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"9493", "BrandName": "cobas c 303" }, { "ID": "8481", "BrandName": "cobas c 503" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSE

FREFN‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine
to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

mmol/L

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

mg/dL

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 65 mg/dL

45‑65 mg/dL

< 45 mg/dL

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 55 mg/dL

35‑55 mg/dL

< 35 mg/dL

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid: No significant interference from ascorbic acid up to a concentration of 2.84 mmol/L (50 mg/dL).

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet for information. For further instructions refer to the operator’s manual.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

08057877190

HDL‑Cholesterol Gen.4 (700 tests)

System‑ID 2071 002

cobas c 303, cobas c 503

Materials required (but not provided):

12172623122

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 20424

05117003190

PreciControl ClinChem Multi 1 (20 x 5 mL)

Code 20391

05947626190

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 20391

05117216190

PreciControl ClinChem Multi 2 (20 x 5 mL)

Code 20392

05947774190

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 20392

08063494190

Diluent NaCl 9 % (123 mL)

System‑ID 2906 001

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 20710

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

Test definition

Reporting time

10 min

Wavelength (sub/main)

700/600 nm

Reagent pipetting

Diluent (H2O)

R1

80 µL

R3

27 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

1.6 µL

Decreased

8.0 µL

10 µL

90 µL

Increased

1.6 µL

For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

12 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

Automatic full calibration
- after reagent lot change

Full calibration
- as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.

Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). Results for repeatability and intermediate precision were obtained on the cobas c 503 analyzer.

Repeatability

Mean

mmol/L

SD

mmol/L

CV

%

PCCC1d)

0.764

0.00290

0.4

PCCC2e)

1.45

0.00690

0.5

Human serum 1

0.148

0.00177

1.2

Human serum 2

1.07

0.00512

0.5

Human serum 3

1.49

0.00673

0.5

Human serum 4

1.92

0.00715

0.4

Human serum 5

3.53

0.0152

0.4

Intermediate precision

Mean

mmol/L

SD

mmol/L

CV

%

PCCC1

FREFPreciControl ClinChem Multi 1

0.760

0.00630

0.8

PCCC2

FREFPreciControl ClinChem Multi 2

1.44

0.00974

0.7

Human serum 1

0.148

0.00229

1.5

Human serum 2

1.07

0.00708

0.7

Human serum 3

1.49

0.0105

0.7

Human serum 4

1.92

0.0145

0.8

Human serum 5

3.54

0.0249

0.7

The data obtained on cobas c 503 analyzer(s) are representative for cobas c 303 analyzer(s).

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).

Sample size (n) = 70

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.001x - 0.0175 mmol/L

y = 1.012x - 0.0274 mmol/L

τ = 0.976

r = 1.000

The sample concentrations were between 0.110 and 3.57 mmol/L.

HDL‑cholesterol values for human serum and plasma samples obtained on a cobas c 303 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).

Sample size (n) = 70

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.011x - 0.0242 mmol/L

y = 1.028x - 0.0389 mmol/L

τ = 0.977

r = 1.000

The sample concentrations were between 0.110 and 3.57 mmol/L.

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSO

FREF 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid
buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R3

Bis-Tris

FREFBis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane
buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

R1 is in position B and R3 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.

Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.

Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.

Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 12 weeks.

Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -70 °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0108057877190c503", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "3", "DocumentObjectID": "FF0000000477A00E", "DocumentOriginID": "FF000000032C180E", "MaterialNumbers": [ "08057877190" ], "InstrumentReferences": [ { "ID": "8481", "BrandName": "cobas c 503" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSE

FREFN‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine
to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

mmol/L

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

mg/dL

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 65 mg/dL

45‑65 mg/dL

< 45 mg/dL

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 55 mg/dL

35‑55 mg/dL

< 35 mg/dL

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid: No significant interference from ascorbic acid up to a concentration of 2.84 mmol/L (50 mg/dL).

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on Roche/Hitachi cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet for information. For further instructions refer to the operator’s manual.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

08057877 190

HDL‑Cholesterol Gen.4 (700 tests)

System‑ID 2071 001

Roche/Hitachi cobas c 503

Materials required (but not provided):

12172623 122

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 20424

05117003 190

PreciControl ClinChem Multi 1 (20 x 5 mL)

Code 20391

05947626 190

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 20391

05117216 190

PreciControl ClinChem Multi 2 (20 x 5 mL)

Code 20392

05947774 190

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 20392

08063494 190

Diluent NaCl 9 % (123 mL)

System‑ID 2906 001

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 20710

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

Test definition

Reporting time

10 min

Wavelength (sub/main)

700/600 nm

Reagent pipetting

Diluent (H2O)

R1

78 µL

R3

26 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

1.3 µL

Decreased

2.6 µL

20 µL

60 µL

Increased

1.3 µL

For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

12 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

Automatic full calibration
- after reagent lot change

Full calibration
- as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.

Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained:

Repeatability

Mean

mmol/L

SD

mmol/L

CV

%

PCCC1d)

0.773

0.00427

0.6

PCCC2e)

2.36

0.0128

0.5

Human serum 1

0.147

0.00161

1.1

Human serum 2

1.07

0.00500

0.5

Human serum 3

1.54

0.00645

0.4

Human serum 4

1.94

0.00715

0.4

Human serum 5

3.08

0.0127

0.4

Intermediate precision

Mean

mmol/L

SD

mmol/L

CV

%

PCCC1

FREFPreciControl ClinChem Multi 1

0.773

0.00645

0.8

PCCC2

FREFPreciControl ClinChem Multi 2

2.36

0.0299

1.3

Human serum 1

0.147

0.00214

1.5

Human serum 2

1.07

0.00791

0.7

Human serum 3

1.54

0.0102

0.7

Human serum 4

1.94

0.0132

0.7

Human serum 5

3.10

0.0218

0.7

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a Roche/Hitachi cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a Roche/Hitachi cobas c 501 analyzer (x).

Sample size (n) = 94

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.014x - 0.00292 mmol/L

y = 1.020x - 0.0114 mmol/L

τ = 0.989

r = 1.000

The sample concentrations were between 0.160 and 3.76 mmol/L.

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSO

FREF 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid
buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R3

Bis-Tris

FREFBis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane
buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

R1 is in position B and R3 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.

Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.

Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.

Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 12 weeks.

Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -70 °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0208057877190c503", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "1", "DocumentObjectID": "FF00000003D6480E", "DocumentOriginID": "FF00000003D6470E", "MaterialNumbers": [ "08057877190" ], "InstrumentReferences": [ { "ID": "8481", "BrandName": "cobas c 503" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSE

FREFNethylN(3methylphenyl)N’succinylethylenediamine
to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

3.09‑150 mg/dL (0.08‑3.88 mmol/L)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 3.09 mg/dL (0.08 mmol/L)

Limit of Detection

= 3.09 mg/dL (0.08 mmol/L)

Limit of Quantitation

= 3.09 mg/dL (0.08 mmol/L)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation for HDL‑C is 3.09 mg/dL determined in accordance with the guidelines in CLSI document EP17-A2, based on a minimum of 48 determinations, and a total error goal of 20 % calculated using RMS error model.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 38.7 mg/dL (1 mmol/L).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 60 mg/dL (1026 µmol/L).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 1200 mg/dL or 745 µmol/L).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 1200 mg/dL or 13.7 mmol/L. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid: No significant interference from ascorbic acid up to a concentration of 50 mg/dL (2.84 mmol/L).

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 450 mg/L (2.76 mmol/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet for information. For further instructions refer to the operator’s manual.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

08057877 190

HDL‑Cholesterol Gen.4 (700 tests)

System‑ID 2071 001

cobas c 503

12172623 160

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 20424

05947626 160

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 20391

05947774 160

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 20392

08063494 190

Diluent NaCl 9 % (123 mL)

System‑ID 2906 001

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 20710

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

Test definition

Reporting time

10 min

Wavelength (sub/main)

700/600 nm

Reagent pipetting

Diluent (H2O)

R1

78 µL

R3

26 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

1.3 µL

Decreased

2.6 µL

20 µL

60 µL

Increased

1.3 µL

For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

12 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

Automatic full calibration
- after reagent lot change

Full calibration
- as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.

Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained:

Repeatability

Mean

mg/dL (mmol/L)

SD

mg/dL (mmol/L)

CV

%

PCCC1d)

29.9 (0.773)

0.165 (0.00427)

0.6

PCCC2e)

91.2 (2.36)

0.495 (0.0128)

0.5

Human serum 1

5.68 (0.147)

0.0622 (0.00161)

1.1

Human serum 2

41.4 (1.07)

0.193 (0.00500)

0.5

Human serum 3

59.5 (1.54)

0.249 (0.00645)

0.4

Human serum 4

75.0 (1.94)

0.276 (0.00715)

0.4

Human serum 5

119 (3.08)

0.491 (0.0127)

0.4

Intermediate precision

Mean

mg/dL (mmol/L)

SD

mg/dL (mmol/L)

CV

%

PCCC1

FREFPreciControl ClinChem Multi 1

29.9 (0.773)

0.249 (0.00645)

0.8

PCCC2

FREFPreciControl ClinChem Multi 2

91.2 (2.36)

1.16 (0.0299)

1.3

Human serum 1

5.68 (0.147)

0.0827 (0.00214)

1.5

Human serum 2

41.4 (1.07)

0.306 (0.00791)

0.7

Human serum 3

59.5 (1.54)

0.394 (0.0102)

0.7

Human serum 4

75.0 (1.94)

0.510 (0.0132)

0.7

Human serum 5

120 (3.10)

0.843 (0.0218)

0.7

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).

Sample size (n) = 94

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.014x - 0.113 mg/dL

y = 1.020x - 0.441 mg/dL

τ = 0.989

r = 1.000

The sample concentrations were between 6.19 and 145 mg/dL (0.160 and 3.76 mmol/L).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSO

FREF 2HydroxyNtris(hydroxymethyl)methyl3aminopropanesulfonic acid
buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R3

Bis-Tris

FREFBis(2hydroxyethyl)iminotris(hydroxymethyl)methane
buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

R1 is in position B and R3 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory reagents.
Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.

For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 12 weeks.

Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0208057877190c503", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "3", "DocumentObjectID": "FF00000005ED290E", "DocumentOriginID": "FF00000003D6480E", "MaterialNumbers": [ "08057877190" ], "InstrumentReferences": [ { "ID": "9493", "BrandName": "cobas c 303" }, { "ID": "8481", "BrandName": "cobas c 503" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSE

FREFN‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine
to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

3.09‑150 mg/dL (0.08‑3.88 mmol/L)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 3.09 mg/dL (0.08 mmol/L)

Limit of Detection

= 3.09 mg/dL (0.08 mmol/L)

Limit of Quantitation

= 3.09 mg/dL (0.08 mmol/L)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation for HDL‑C is 3.09 mg/dL determined in accordance with the guidelines in CLSI document EP17-A2, based on a minimum of 48 determinations, and a total error goal of 20 % calculated using RMS error model.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 38.7 mg/dL (1 mmol/L).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 60 mg/dL (1026 µmol/L).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 1200 mg/dL or 745 µmol/L).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 1200 mg/dL or 13.7 mmol/L. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid: No significant interference from ascorbic acid up to a concentration of 50 mg/dL (2.84 mmol/L).

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 450 mg/L (2.76 mmol/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SCCS Method Sheet. For further instructions refer to the operator’s manual.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

08057877190

HDL‑Cholesterol Gen.4 (700 tests)

System‑ID 2071 002

cobas c 303, cobas c 503

Materials required (but not provided):

12172623160

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 20424

05947626160

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 20391

05947774160

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 20392

08063494190

Diluent NaCl 9 % (123 mL)

System‑ID 2906 001

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 20710

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

Test definition

Reporting time

10 min

Wavelength (sub/main)

700/600 nm

Reagent pipetting

Diluent (H2O)

R1

80 µL

R3

27 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

1.6 µL

Decreased

8.0 µL

10 µL

90 µL

Increased

1.6 µL

For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

12 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

Automatic full calibration
- after reagent lot change

Full calibration
- as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.

Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). Results for repeatability and intermediate precision were obtained on the cobas c 503 analyzer.

Repeatability

Mean

mg/dL (mmol/L)

SD

mg/dL (mmol/L)

CV

%

PCCC1d)

29.5 (0.764)

0.112 (0.00290)

0.4

PCCC2e)

56.0 (1.45)

0.267 (0.00690)

0.5

Human serum 1

5.72 (0.148)

0.0684 (0.00177)

1.2

Human serum 2

41.4 (1.07)

0.198 (0.00512)

0.5

Human serum 3

57.6 (1.49)

0.260 (0.00673)

0.5

Human serum 4

74.2 (1.92)

0.276 (0.00715)

0.4

Human serum 5

136 (3.53)

0.588 (0.0152)

0.4

Intermediate precision

Mean

mg/dL (mmol/L)

SD

mg/dL (mmol/L)

CV

%

PCCC1

FREFPreciControl ClinChem Multi 1

29.4 (0.760)

0.244 (0.00630)

0.8

PCCC2

FREFPreciControl ClinChem Multi 2

55.7 (1.44)

0.377 (0.00974)

0.7

Human serum 1

5.72 (0.148)

0.0885 (0.00229)

1.5

Human serum 2

41.4 (1.07)

0.274 (0.00708)

0.7

Human serum 3

57.6 (1.49)

0.406 (0.0105)

0.7

Human serum 4

74.2 (1.92)

0.561 (0.0145)

0.8

Human serum 5

137 (3.54)

0.963 (0.0249)

0.7

The data obtained on cobas c 503 analyzer(s) are representative for cobas c 303 analyzer(s).

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a cobas c 503 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).

Sample size (n) = 70

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.001x - 0.677 mg/dL

y = 1.012x - 1.06 mg/dL

τ = 0.976

r = 1.000

The sample concentrations were between 4.25 and 138 mg/dL (0.110 and 3.57 mmol/L).

HDL‑cholesterol values for human serum and plasma samples obtained on a cobas c 303 analyzer (y) were compared with those determined using the corresponding reagent on a cobas c 501 analyzer (x).

Sample size (n) = 70

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.011x - 0.936 mg/dL

y = 1.028x - 1.50 mg/dL

τ = 0.977

r = 1.000

The sample concentrations were between 4.25 and 138 mg/dL (0.110 and 3.57 mmol/L).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSO

FREF 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid
buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R3

Bis-Tris

FREFBis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane
buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

R1 is in position B and R3 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.

Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.

Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.

Safety data sheet available for professional user on request.

For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 12 weeks.

Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0107528582190c701", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "4", "DocumentObjectID": "FF00000003498A0E", "DocumentOriginID": "FF00000001BF630E", "MaterialNumbers": [ "07528582190", "07528582214" ], "InstrumentReferences": [ { "ID": "2492", "BrandName": "cobas c 702" }, { "ID": "310", "BrandName": "cobas c 701" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

(> 65 mg/dL)

(45‑65 mg/dL)

(< 45 mg/dL)

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

(> 55 mg/dL)

(35‑55 mg/dL)

(< 35 mg/dL)

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid up to 2.84 mmol/L (50 mg/dL) does not interfere.

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on Roche/Hitachi cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link, manual input is required in certain cases. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SmpCln1+2/SCCS Method Sheet and for further instructions refer to the operator’s manual.

Where required, special wash/carry‑over evasion programming must be implemented prior to reporting results with this test.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

07528582 190*

HDL‑Cholesterol Gen.4 (500 tests)

System‑ID 05 7589 4

Roche/Hitachi cobas c 701/702

07528582 214*

HDL‑Cholesterol Gen.4 (500 tests)

System‑ID 05 7589 4

Roche/Hitachi cobas c 701/702

12172623 122

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 424

05117003 190

PreciControl ClinChem Multi 1 (20 x 5 mL)

Code 391

05947626 190

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 391

05117216 190

PreciControl ClinChem Multi 2 (20 x 5 mL)

Code 392

05947774 190

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 392

05172152 190

Diluent NaCl 9 % (119 mL)

System ID 08 6869 3

* Some kits shown may not be available in all countries.

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 8454

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

cobas c 701/702 test definition

Assay type

2‑Point End

Reaction time / Assay points

10/18‑38

Wavelength (sub/main)

700/600 nm

Reaction direction

Increase

Units

mmol/L (mg/dL)

Reagent pipetting

Diluent (H2O)

R1

120 µL

R3

40 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

2.4 µL

Decreased

12 µL

15 µL

135 µL

Increased

2.4 µL

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

HDLC4

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

4 weeks

On‑board on the Reagent Manager:

24 hours

Diluent NaCl 9 %

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

4 weeks

On‑board on the Reagent Manager:

24 hours

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

2‑point calibration
• after reagent lot change
• as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (4 aliquots per run, 1 run per day, 21 days). The following results were obtained:

Repeatability

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.70 (27.1)

0.004 (0.16)

0.6

PCCC Multi 2

1.67 (64.6)

0.01 (0.39)

0.4

Human serum 1

0.24 (9.28)

0.002 (0.08)

0.9

Human serum 2

1.01 (39.1)

0.01 (0.39)

0.6

Human serum 3

1.47 (56.8)

0.01 (0.39)

0.5

Human serum 4

1.96 (75.8)

0.01 (0.39)

0.4

Human serum 5

3.53 (137)

0.01 (0.39)

0.3

Intermediate precision

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.70 (27.1)

0.01 (0.39)

1.4

PCCC Multi 2

1.67 (64.6)

0.03 (1.16)

1.6

Human serum 1

0.24 (9.28)

0.003 (0.12)

1.4

Human serum 2

1.01 (39.1)

0.02 (0.77)

1.8

Human serum 3

1.47 (56.8)

0.02 (0.77)

1.7

Human serum 4

1.96 (75.8)

0.02 (0.77)

1.2

Human serum 5

3.53 (137)

0.04 (1.55)

1.2

PCCC = PreciControl ClinChem

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a Roche/Hitachi cobas c 701 analyzer (y) were compared with those determined using the corresponding reagent on a Roche/Hitachi cobas c 501 analyzer (x).

Sample size (n) = 59

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 0.994x - 0.032 mmol/L

y = 0.987x - 0.020 mmol/L

τ = 0.994

r = 1.000

The sample concentrations were between 0.12 and 3.74 mmol/L (4.64 and 145 mg/dL).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSOb) buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R3

Bis-Trisc) buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

b) 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid

c) Bis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane

R1 is in position B and R3 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory reagents.
Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -70 °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0107528582190c701", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "5", "DocumentObjectID": "FF0000000477430E", "DocumentOriginID": "FF00000003498A0E", "MaterialNumbers": [ "07528582190", "07528582214" ], "InstrumentReferences": [ { "ID": "2492", "BrandName": "cobas c 702" }, { "ID": "310", "BrandName": "cobas c 701" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

(> 65 mg/dL)

(45‑65 mg/dL)

(< 45 mg/dL)

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

(> 55 mg/dL)

(35‑55 mg/dL)

(< 35 mg/dL)

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid up to 2.84 mmol/L (50 mg/dL) does not interfere.

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on Roche/Hitachi cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link, manual input is required in certain cases. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SmpCln1+2/SCCS Method Sheet and for further instructions refer to the operator’s manual.

Where required, special wash/carry‑over evasion programming must be implemented prior to reporting results with this test.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

07528582 190*

HDL‑Cholesterol Gen.4 (500 tests)

System‑ID 05 7589 4

Roche/Hitachi cobas c 701/702

07528582 214*

HDL‑Cholesterol Gen.4 (500 tests)

System‑ID 05 7589 4

Roche/Hitachi cobas c 701/702

Materials required (but not provided):

12172623 122

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 424

05117003 190

PreciControl ClinChem Multi 1 (20 x 5 mL)

Code 391

05947626 190

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 391

05117216 190

PreciControl ClinChem Multi 2 (20 x 5 mL)

Code 392

05947774 190

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 392

05172152 190

Diluent NaCl 9 % (119 mL)

System ID 08 6869 3

* Some kits shown may not be available in all countries.

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 8454

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

cobas c 701/702 test definition

Assay type

2‑Point End

Reaction time / Assay points

10/18‑38

Wavelength (sub/main)

700/600 nm

Reaction direction

Increase

Units

mmol/L (mg/dL)

Reagent pipetting

Diluent (H2O)

R1

120 µL

R3

40 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

2.4 µL

Decreased

12 µL

15 µL

135 µL

Increased

2.4 µL

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

HDLC4

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

4 weeks

On‑board on the Reagent Manager:

24 hours

Diluent NaCl 9 %

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

4 weeks

On‑board on the Reagent Manager:

24 hours

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

2‑point calibration
• after reagent lot change
• as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (4 aliquots per run, 1 run per day, 21 days). The following results were obtained:

Repeatability

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.70 (27.1)

0.004 (0.16)

0.6

PCCC Multi 2

1.67 (64.6)

0.01 (0.39)

0.4

Human serum 1

0.24 (9.28)

0.002 (0.08)

0.9

Human serum 2

1.01 (39.1)

0.01 (0.39)

0.6

Human serum 3

1.47 (56.8)

0.01 (0.39)

0.5

Human serum 4

1.96 (75.8)

0.01 (0.39)

0.4

Human serum 5

3.53 (137)

0.01 (0.39)

0.3

Intermediate precision

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.70 (27.1)

0.01 (0.39)

1.4

PCCC Multi 2

1.67 (64.6)

0.03 (1.16)

1.6

Human serum 1

0.24 (9.28)

0.003 (0.12)

1.4

Human serum 2

1.01 (39.1)

0.02 (0.77)

1.8

Human serum 3

1.47 (56.8)

0.02 (0.77)

1.7

Human serum 4

1.96 (75.8)

0.02 (0.77)

1.2

Human serum 5

3.53 (137)

0.04 (1.55)

1.2

PCCC = PreciControl ClinChem

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a Roche/Hitachi cobas c 701 analyzer (y) were compared with those determined using the corresponding reagent on a Roche/Hitachi cobas c 501 analyzer (x).

Sample size (n) = 59

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 0.994x - 0.032 mmol/L

y = 0.987x - 0.020 mmol/L

τ = 0.994

r = 1.000

The sample concentrations were between 0.12 and 3.74 mmol/L (4.64 and 145 mg/dL).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSOb) buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R3

Bis-Trisc) buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

b) 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid

c) Bis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane

R1 is in position B and R3 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.

Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.

Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.

Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -70 °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0207528604190_USA", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "1", "DocumentObjectID": "FF0000000330FE0E", "DocumentOriginID": "FF0000000330FE0E", "MaterialNumbers": [ "07528604190" ], "InstrumentReferences": [ { "ID": "307", "BrandName": "cobas c 111" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on cobas c 111 systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

3.09‑150 mg/dL (0.08‑3.88 mmol/L)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 3.09 mg/dL (0.08 mmol/L)

Limit of Detection

= 3.09 mg/dL (0.08 mmol/L)

Limit of Quantitation

= 3.09 mg/dL (0.08 mmol/L)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation for HDL‑cholesterol is 3.09 mg/dL determined in accordance with the guidelines in CLSI document EP17‑A2, based on a minimum of 48 determinations, and a total error goal of 20 % calculated using RMS error model.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 38.7 mg/dL (1 mmol/L).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 60 mg/dL or 1026 µmol/L).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 1200 mg/dL or 745 µmol/L).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 1200 mg/dL or 13.7 mmol/L. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid: No significant interference from ascorbic acid up a concentration of 50 mg/dL (2.84 mmol/L).

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to a concentration of 450 mg/L (2.76 mmol/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on the cobas c 111 analyzer. For information about test combinations requiring special wash steps, please refer to the latest version of the carry over evasion list found with the CLEAN Method Sheet and the operator’s manual for further instructions.
Where required, special wash/carry-over evasion programming must be implemented prior to reporting results with this test.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which kit(s) can be used

07528604 190

HDL‑Cholesterol Gen.4 (2 x 100 tests)

cobasc 111

12172623 160

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 424

05947626 160

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 391

05947774 160

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 392

04774230 190

Diluent NaCl 9 %

Code 951

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 454

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

cobas c 111 test definition

Measuring mode

Absorbance

Abs. calculation mode

Endpoint

Reaction direction

Increase

Wavelength A/B

583/659 nm

Calc. first/last

16/37

Unit

mmol/L

Reaction mode

R1‑S‑SR

Pipetting parameters

Diluent (H2O)

R1

120 µL

Sample

2.5 µL

7 µL

SR

40 µL

Total volume

169.5 µL

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

HDLC4

Shelf life at 2‑8 °C:

See expiration date on reagent.

On‑board in use and refrigerated on the analyzer:

3 weeks

Diluent NaCl 9 %

Shelf life at 2‑8 °C:

See expiration date on reagent.

On‑board in use and refrigerated on the analyzer:

4 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

C.f.a.s. Lipids

Deionized water is used automatically by the instrument as the zero calibrator.

Calibration mode

Linear regression

Calibration frequency

Each lot and as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (4 aliquots per run, 1 run per day, 21 days). The following results were obtained:

Repeatability

Mean

mg/dL (mmol/L)

SD

mg/dL (mmol/L)

CV

%

PCCC Multi 1

26.6 (0.69)

0.40 (0.01)

1.5

PCCC Multi 2

66.4 (1.72)

1.05 (0.03)

1.6

Human serum 1

8.92 (0.23)

0.20 (0.01)

2.3

Human serum 2

38.1 (0.99)

0.61 (0.02)

1.6

Human serum 3

56.4 (1.46)

0.73 (0.02)

1.3

Human serum 4

74.3 (1.92)

1.11 (0.03)

1.5

Human serum 5

134 (3.47)

1.76 (0.05)

1.3

Intermediate precision

Mean

mg/dL (mmol/L)

SD

mg/dL (mmol/L)

CV

%

PCCC Multi 1

26.6 (0.69)

0.51 (0.01)

1.9

PCCC Multi 2

66.4 (1.72)

1.42 (0.04)

2.1

Human serum 1

8.92 (0.23)

0.22 (0.01)

2.5

Human serum 2

38.1 (0.99)

0.71 (0.02)

1.8

Human serum 3

56.4 (1.46)

1.02 (0.03)

1.8

Human serum 4

74.3 (1.92)

1.25 (0.03)

1.7

Human serum 5

134 (3.47)

2.61 (0.07)

2.0

PCCC = PreciControl ClinChem

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL-cholesterol values for human serum samples obtained on a cobas c 111 analyzer (y) were compared with those determined using the corresponding reagent on a COBAS INTEGRA 400 plus analyzer (x).

Sample size (n) = 57

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.015x - 0.979 mg/dL

y = 1.006x - 0.314 mg/dL

τ = 0.968

r = 0.999

The sample concentrations were between 4.65 and 145 mg/dL (0.12 and 3.76 mmol/L).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSOb) buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

SR

Bis-Trisc) buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

b) 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid

c) Bis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory reagents.
Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.

For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Sample stability claims were established by experimental data by the manufacturer or based on reference literature and only for the temperatures/time frames as stated in the method sheet. It is the responsibility of the individual laboratory to use all available references and/or its own studies to determine specific stability criteria for its laboratory.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "07529031001", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "2", "DocumentObjectID": "FF00000004774C0E", "DocumentOriginID": "FF00000001BF610E", "MaterialNumbers": [ "07528604190" ], "InstrumentReferences": [ { "ID": "307", "BrandName": "cobas c 111" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on cobas c 111 systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

(> 65 mg/dL)

(45‑65 mg/dL)

(< 45 mg/dL)

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

(> 55 mg/dL)

(35‑55 mg/dL)

(< 35 mg/dL)

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid up to 2.84 mmol/L (50 mg/dL) does not interfere.

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference up to a N‑acetylcysteine concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on the cobas c 111 analyzer. For information about test combinations requiring special wash steps, please refer to the latest version of the carry over evasion list found with the CLEAN Method Sheet and the operator’s manual for further instructions.
Where required, special wash/carry-over evasion programming must be implemented prior to reporting results with this test.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which kit(s) can be used

07528604 190

HDL‑Cholesterol Gen.4 (2 x 100 tests)

cobasc 111

Materials required (but not provided):

12172623 122

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 424

05117003 190

PreciControl ClinChem Multi 1 (20 x 5 mL)

Code 391

05947626 190

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 391

05117216 190

PreciControl ClinChem Multi 2 (20 x 5 mL)

Code 392

05947774 190

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 392

04774230 190

Diluent NaCl 9 %

Code 951

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

HDLC4: ACN 454

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

cobas c 111 test definition

Measuring mode

Absorbance

Abs. calculation mode

Endpoint

Reaction direction

Increase

Wavelength A/B

583/659 nm

Calc. first/last

16/37

Unit

mmol/L

Reaction mode

R1‑S‑SR

Pipetting parameters

Diluent (H2O)

R1

120 µL

Sample

2.5 µL

7 µL

SR

40 µL

Total volume

169.5 µL

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

HDLC4

Shelf life at 2‑8 °C:

See expiration date on reagent.

On‑board in use and refrigerated on the analyzer:

3 weeks

Diluent NaCl 9 %

Shelf life at 2‑8 °C:

See expiration date on reagent.

On‑board in use and refrigerated on the analyzer:

4 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

C.f.a.s. Lipids

Deionized water is used automatically by the instrument as the zero calibrator.

Calibration mode

Linear regression

Calibration frequency

Each lot and as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.
LREFData on file at Roche Diagnostics.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (4 aliquots per run, 1 run per day, 21 days). The following results were obtained:

Repeatability

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.69 (26.6)

0.01 (0.39)

1.5

PCCC Multi 2

1.72 (66.3)

0.03 (1.04)

1.6

Human serum 1

0.23 (8.93)

0.01 (0.19)

2.3

Human serum 2

0.99 (38.2)

0.02 (0.62)

1.6

Human serum 3

1.46 (56.4)

0.02 (0.73)

1.3

Human serum 4

1.92 (74.3)

0.03 (1.12)

1.5

Human serum 5

3.46 (134)

0.05 (1.74)

1.3

Intermediate precision

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.69 (26.6)

0.01 (0.50)

1.9

PCCC Multi 2

1.72 (66.3)

0.04 (1.43)

2.1

Human serum 1

0.23 (8.93)

0.01 (0.23)

2.5

Human serum 2

0.99 (38.2)

0.02 (0.70)

1.8

Human serum 3

1.46 (56.4)

0.03 (1.01)

1.8

Human serum 4

1.92 (74.3)

0.03 (1.24)

1.7

Human serum 5

3.46 (134)

0.07 (2.59)

2.0

PCCC = PreciControl ClinChem

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL-cholesterol values for human serum samples obtained on a cobas c 111 analyzer (y) were compared to those determined using the corresponding reagent on a COBAS INTEGRA 400 plus analyzer (x).

Sample size (n) = 57

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.015x - 0.025 mmol/L

y = 1.006x - 0.008 mmol/L

τ = 0.968

r = 0.999

The sample concentrations were between 0.12 and 3.74 mmol/L (4.64 and 145 mg/dL).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSOb) buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

SR

Bis-Trisc) buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

b) 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid

c) Bis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.

Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.

Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.

Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

LREFData on file at Roche Diagnostics.

7 days at 2‑8 °C

LREFData on file at Roche Diagnostics.

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

LREFData on file at Roche Diagnostics.

7 days at 2‑8 °C

LREFData on file at Roche Diagnostics.

3 months at (-15)‑(-25) °C

LREFData on file at Roche Diagnostics.

18 months at -70 °C

LREFData on file at Roche Diagnostics.

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0107528566190c501", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "3", "DocumentObjectID": "FF0000000212050E", "DocumentOriginID": "FF0000000212050E", "MaterialNumbers": [ "07528566190" ], "InstrumentReferences": [ { "ID": "308", "BrandName": "cobas c 311" }, { "ID": "2324", "BrandName": "cobas c 502" }, { "ID": "309", "BrandName": "cobas c 501" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

(> 65 mg/dL)

(45‑65 mg/dL)

(< 45 mg/dL)

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

(> 55 mg/dL)

(35‑55 mg/dL)

(< 35 mg/dL)

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid up to 2.84 mmol/L (50 mg/dL) does not interfere.

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference from N‑acetylcysteine up to an concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on Roche/Hitachi cobas c systems. The latest version of the carry‑over evasion list can be found with the NaOHD-SMS-SmpCln1+2-SCCS Method Sheets. For further instructions refer to the operator’s manual. cobas c 502 analyzer: All special wash programming necessary for avoiding carry‑over is available via the cobas link, manual input is required in certain cases.

Where required, special wash/carry‑over evasion programming must be implemented prior to reporting results with this test.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

07528566 190

HDL‑Cholesterol Gen.4 (350 tests)

System‑ID 07 7589 4

Roche/Hitachi cobas c 311, cobas c 501/502

12172623 122

Calibrator f.a.s. Lipids (3 x 1 mL)

Code 424

05117003 190

PreciControl ClinChem Multi 1 (20 x 5 mL)

Code 391

05947626 190

PreciControl ClinChem Multi 1 (4 x 5 mL)

Code 391

05117216 190

PreciControl ClinChem Multi 2 (20 x 5 mL)

Code 392

05947774 190

PreciControl ClinChem Multi 2 (4 x 5 mL)

Code 392

04489357 190

Diluent NaCl 9 % (50 mL)

System ID 07 6869 3

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

For cobas c 311/501 analyzers:

HDLC4: ACN 454

For cobas c 502 analyzer:

HDLC4: ACN 8454

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

cobas c 311 test definition

Assay type

2‑Point End

Reaction time / Assay points

10/6‑33

Wavelength (sub/main)

700/600 nm

Reaction direction

Increase

Units

mmol/L (mg/dL)

Reagent pipetting

Diluent (H2O)

R1

120 µL

R2

40 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

2.4 µL

Decreased

12 µL

15 µL

135 µL

Increased

2.4 µL

cobas c 501/502 test definition

Assay type

2‑Point End

Reaction time / Assay points

10/10‑47

Wavelength (sub/main)

700/600 nm

Reaction direction

Increase

Units

mmol/L (mg/dL)

Reagent pipetting

Diluent (H2O)

R1

120 µL

R2

40 µL

Sample volumes

Sample

Sample dilution

Sample

Diluent (NaCl)

Normal

2.4 µL

Decreased

12 µL

15 µL

135 µL

Increased

2.4 µL

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

HDLC4

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

12 weeks

Diluent NaCl 9 %

Shelf life at 2‑8 °C:

See expiration date on cobas c pack label.

On‑board in use and refrigerated on the analyzer:

12 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrators

S1: H2O

S2: C.f.a.s. Lipids

Calibration mode

Linear

Calibration frequency

2‑point calibration
• after reagent lot change
• as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (4 aliquots per run, 1 run per day, 21 days). The following results were obtained:

Repeatability

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.73 (28.2)

0.004 (0.15)

0.6

PCCC Multi 2

1.76 (68.0)

0.01 (0.39)

0.6

Human serum 1

0.25 (9.67)

0.004 (0.15)

1.8

Human serum 2

1.05 (40.6)

0.01 (0.39)

0.7

Human serum 3

1.53 (59.1)

0.01 (0.39)

0.5

Human serum 4

2.05 (79.3)

0.01 (0.39)

0.6

Human serum 5

3.66 (141)

0.02 (0.77)

0.6

Intermediate precision

Mean

mmol/L (mg/dL)

SD

mmol/L (mg/dL)

CV

%

PCCC Multi 1

0.73 (28.2)

0.01 (0.27)

1.0

PCCC Multi 2

1.72 (66.5)

0.02 (0.77)

1.4

Human serum 1

0.25 (9.67)

0.01 (0.19)

2.2

Human serum 2

1.05 (40.6)

0.01 (0.39)

0.8

Human serum 3

1.53 (59.1)

0.01 (0.39)

0.7

Human serum 4

2.05 (79.3)

0.02 (0.77)

0.8

Human serum 5

3.66 (141)

0.03 (1.16)

0.8

PCCC = PreciControl ClinChem

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a Roche/Hitachi cobas c 701 analyzer (y) were compared with those determined using the corresponding reagent on a Roche/Hitachi cobas c 501 analyzer (x).

Sample size (n) = 59

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.006x + 0.032 mmol/L

y = 1.012x + 0.021 mmol/L

τ = 0.994

r = 1.000

The sample concentrations were between 0.11 and 3.69 mmol/L (4.25 and 143 mg/dL).

HDL‑cholesterol values for human serum and plasma samples obtained on a COBAS INTEGRA 400 plus analyzer (y) were compared with those determined using the corresponding reagent on a Roche/Hitachi cobas c 501 analyzer (x).

Sample size (n) = 118

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 0.980x + 0.013 mmol/L

y = 0.988x + 0.001 mmol/L

τ = 0.973

r = 0.998

The sample concentrations were between 0.08 and 3.83 mmol/L (3.09 and 148 mg/dL).

HDL‑cholesterol values for human serum samples obtained on a Roche/Hitachi cobas c 501 analyzer (y) were compared with those determined using the HDL Ultra Cholesterol Reagent on a Roche/Hitachi 917 analyzer (x).

Sample size (n) = 111

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 0.957x - 0.024 mmol/L

y = 0.961x - 0.033 mmol/L

τ = 0.944

r = 0.995

The sample concentrations were between 0.13 and 3.86 mmol/L (5.03 and 149 mg/dL).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSOb) buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

R2

Bis-Trisc) buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

b) 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid

c) Bis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane

R1 is in position B and R2 is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use.
Exercise the normal precautions required for handling all laboratory reagents.
Disposal of all waste material should be in accordance with local guidelines.
Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

See the limitations and interferences section for details about possible sample interferences.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

7 days at 2‑8 °C

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

7 days at 2‑8 °C

3 months at (-15)‑(-25) °C

18 months at -70 °C

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0107528566190COIN", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "3", "DocumentObjectID": "FF0000000477130E", "DocumentOriginID": "FF00000001BF640E", "MaterialNumbers": [ "07528566190" ], "InstrumentReferences": [ { "ID": "302", "BrandName": "COBAS INTEGRA 400 plus" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on COBAS INTEGRA systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

HDL‑cholesterol esters + H2O

CHER

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

HDL‑cholesterol + O2

CHOD

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

2 H2O2 + 4‑amino‑antipyrine + EMSE + H+ + H2O

Peroxidase

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

(> 65 mg/dL)

(45‑65 mg/dL)

(< 45 mg/dL)

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

(> 55 mg/dL)

(35‑55 mg/dL)

(< 35 mg/dL)

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:
LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (approximate conjugated and unconjugated bilirubin concentration: 1026 µmol/L or 60 mg/dL).

Hemolysis:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an H index of 1200 (approximate hemoglobin concentration: 745 µmol/L or 1200 mg/dL).

Lipemia (Intralipid):

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an L index of 2000. No significant interference from native triglycerides up to 13.7 mmol/L or 1200 mg/dL. There is poor correlation between the L index (corresponds to turbidity) and triglycerides concentration.

Other: Elevated concentrations of free fatty acids and denatured proteins may cause falsely elevated HDL‑cholesterol results.

Ascorbic acid up to 2.84 mmol/L (50 mg/dL) does not interfere.

Abnormal liver function affects lipid metabolism; consequently, HDL and LDL results are of limited diagnostic value. In some patients with abnormal liver function, the HDL‑cholesterol result may significantly differ from the DCM (designated comparison method) result due to the presence of lipoproteins with abnormal lipid distribution.

LREFDati F, Metzmann E. Proteins Laboratory Testing and Clinical Use, Verlag: DiaSys; 1. Auflage (September 2005), page 242-243; ISBN-10: 3000171665.

Drugs: No interference was found at therapeutic concentrations using common drug panels.

LREFBreuer J. Report on the Symposium "Drug effects in Clinical Chemistry Methods". Eur J Clin Chem Clin Biochem 1996;34:385-386.
,
LREFSonntag O, Scholer A. Drug interference in clinical chemistry: recommendation of drugs and their concentrations to be used in drug interference studies. Ann Clin Biochem 2001;38:376-385.

Statins (Simvastatin) and fibrates (Bezafibrate) tested at therapeutic concentration ranges did not interfere.

N‑acetylcysteine: No significant interference up to a N‑acetylcysteine concentration of 2.76 mmol/L (450 mg/L).

Acetaminophen intoxications are frequently treated with N‑acetylcysteine. N‑acetylcysteine at the therapeutic concentration when used as an antidote and the acetaminophen metabolite N‑acetyl‑p‑benzoquinone imine (NAPQI) independently may cause falsely low HDL‑cholesterol results.

Metamizole: Venipuncture should be performed prior to the administration of metamizole. Venipuncture immediately after or during the administration of metamizole may lead to falsely low results.

In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.

LREFBakker AJ, Mücke M. Gammopathy interference in clinical chemistry assays: mechanisms, detection and prevention. Clin Chem Lab Med 2007;45(9):1240-1243.

For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.

ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on COBAS INTEGRA analyzers. Refer to the CLEAN Method Sheet for further instructions and for the latest version of the Extra wash cycle list.
Where required, special wash/carry-over evasion programming must be implemented prior to reporting results with this test.

", "Language": "en" }, { "Name": "OrderInformation", "Value": "

OrderInformation (CC Reagents - cobas + Integra)

Order information

Analyzer(s) on which cobas c pack(s) can be used

07528566 190

HDL‑Cholesterol Gen.4 (350 tests)

System‑ID 07 7589 4

COBAS INTEGRA 400 plus

Materials required (but not provided):

12172623 122

Calibrator f.a.s. Lipids (3 x 1 mL)

System‑ID 07 6570 8

05117003 190

PreciControl ClinChem Multi 1 (20 x 5 mL)

System‑ID 07 7469 3

05947626 190

PreciControl ClinChem Multi 1 (4 x 5 mL)

System‑ID 07 7469 3

05117216 190

PreciControl ClinChem Multi 2 (20 x 5 mL)

System‑ID 07 7470 7

05947774 190

PreciControl ClinChem Multi 2 (4 x 5 mL)

System‑ID 07 7470 7

20756350 322

Diluent NaCl 9 % (6 x 22 mL)

System ID 07 5635 0

", "Language": "en" }, { "Name": "SystemInformation", "Value": "

System information

Test HDLC4, test ID 0‑389

", "Language": "en" }, { "Name": "Handling", "Value": "

Reagent handling

Ready for use

The intrinsic color of the reagent does not interfere with the test.

", "Language": "en" }, { "Name": "TestDefinition", "Value": "

Application for serum and plasma

COBAS INTEGRA 400 plus test definition

Measuring mode

Absorbance

Abs. calculation mode

Endpoint

Reaction mode

R1-S-SR

Reaction direction

Increase

Wavelength A/B

583/800 nm

Calc. first/last

33/69

Unit

mmol/L

Pipetting parameters

Diluent (H2O)

R1

120 µL

Sample

2.5 µL

7 µL

SR

40 µL

Total volume

169.5 µL

", "Language": "en" }, { "Name": "StorageStability", "Value": "

Storage and stability

HDLC4

Shelf life at 2‑8 °C

See expiration date on cobas c pack label

On‑board in use at 10‑15 °C

12 weeks

Diluent NaCl 9 %

Shelf life at 2‑8 °C

See expiration date on cobas c pack label

On‑board in use at 10‑15 °C

4 weeks

", "Language": "en" }, { "Name": "Calibration", "Value": "

Calibration

Calibrator

C.f.a.s. Lipids

Use deionized water as zero calibrator.

Calibration mode

Linear regression

Calibration replicate

Duplicate recommended

Calibration interval

Each lot and as required following quality control procedures

Calibration interval may be extended based on acceptable verification of calibration by the laboratory.

Traceability: This method has been standardized against the designated CDC reference method (ultracentrifugation method).

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
The standardization meets the requirements of the “HDL Cholesterol Method Evaluation Protocol for Manufacturers” of the US National Reference System for Cholesterol, CRMLN (Cholesterol Reference Method Laboratory Network), November 1994.
LREFData on file at Roche Diagnostics.

", "Language": "en" }, { "Name": "Limitations", "Value": "", "Language": "en" }, { "Name": "PerformanceData", "Value": "

Specific performance data

Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.

", "Language": "en" }, { "Name": "Precision", "Value": "

Precision

Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (4 aliquots per run, 1 run per day, 21 days). The following results were obtained:

Repeatability

Mean
mmol/L (mg/dL)

SD
mmol/L (mg/dL)

CV
%

PCCC Multi 1

0.71 (27.5)

0.01 (0.27)

1.0

PCCC Multi 2

1.73 (66.9)

0.02 (0.85)

1.2

Human serum 1

0.23 (8.89)

0.004 (0.16)

1.9

Human serum 2

1.00 (38.7)

0.01 (0.43)

1.1

Human serum 3

1.47 (56.8)

0.02 (0.66)

1.2

Human serum 4

1.91 (73.8)

0.02 (0.77)

1.0

Human serum 5

3.44 (133)

0.05 (1.79)

1.3

Intermediate precision

Mean
mmol/L (mg/dL)

SD
mmol/L (mg/dL)

CV
%

PCCC Multi 1

0.70 (27.1)

0.01 (0.43)

1.5

PCCC Multi 2

1.70 (65.7)

0.03 (1.004)

1.6

Human serum 1

0.23 (8.89)

0.01 (0.19)

2.3

Human serum 2

1.00 (38.7)

0.01 (0.54)

1.4

Human serum 3

1.47 (56.8)

0.02 (0.77)

1.4

Human serum 4

1.91 (73.8)

0.03 (0.97)

1.3

Human serum 5

3.44 (133)

0.05 (2.01)

1.5

PCCC = PreciControl ClinChem

", "Language": "en" }, { "Name": "MethodComparison", "Value": "

Method comparison

HDL‑cholesterol values for human serum and plasma samples obtained on a COBAS INTEGRA 400 plus analyzer (y) were compared with those determined using the corresponding reagent on a Roche/Hitachi cobas c 501 analyzer (x).

Sample size (n) = 60

Passing/Bablok

LREFBablok W, Passing H, Bender R, et al. A general regression procedure for method transformation. Application of linear regression procedures for method comparison studies in clinical chemistry, Part III. J Clin Chem Clin Biochem 1988 Nov;26(11):783-790.

Linear regression

y = 1.012x - 0.005 mmol/L

y = 1.010x - 0.005 mmol/L

τ = 0.973

r = 0.999

The sample concentrations were between 0.08 and 3.74 mmol/L (3.09 and 144.58 mg/dL).

", "Language": "en" }, { "Name": "Summary", "Value": "

Summary

High density lipoproteins (HDL) are responsible for the reverse transport of cholesterol from the peripheral cells to the liver. In the liver, cholesterol is transformed to bile acids which are then excreted into the intestine via the biliary tract.
Monitoring of HDL‑cholesterol in serum or plasma is of clinical relevance as the HDL‑cholesterol concentration is important in the assessment of atherosclerotic risk. Elevated HDL‑cholesterol concentrations protect against coronary heart disease (CHD), whereas reduced HDL‑cholesterol concentrations, particularly in conjunction with elevated triglycerides, increase cardiovascular risk.

LREFDominiczak M, McNamara J. The system of Cardiovascular prevention. 103.125; Nauk M, Wiebe D, Warnick G.Measurement of High-Density-Lipoprotein Cholesterol.221.244. In: Handbook of Lipoprotein Testing (eds. Rifai,Warnick and Dominiczak), 2nd edition.

Two cholesterol related variables that are predictive of cardiovascular disease (CVD) have emerged. These are non‑HDL‑cholesterol

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.
,
LREFBoekholdt SM, Arsenault BJ, Mora S, et al. Association of LDL cholesterol, non-HDL cholesterol, and apolipoprotein B levels with risk of cardiovascular events among patients treated with statins: a meta-analysis. JAMA 2012 Mar 28;307(12):1302-9.
,
LREFStone NJ, Robinson JG, Lichtenstein AH, et al. 2013 ACC/AHA guideline on the treatment of blood cholesterol to reduce atherosclerotic cardiovascular risk in adults: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines. J Am Coll Cardiol 2014;63:2889-2934.
(= cholesterol ‑ HDL‑cholesterol) and the rate of cholesterol transfer from the macrophages to HDL, also described as cholesterol efflux capacity.
LREFRohatgi A, Khera A, Berry JD, et al. HDL cholesterol efflux capacity and incident cardiovascular events. N Engl J Med 2014 Dec 18;371(25):2383-93.
Whereas both cholesterol and HDL‑cholesterol can be readily determined with high accuracy, currently, non‑HDL‑cholesterol appears to be best suited for patient management.

A variety of methods are available to determine HDL‑cholesterol, including ultracentrifugation (reference method in combination with cholesterol measurement by the Abell‑Kendall method), electrophoresis, HPLC, precipitation, and direct methods.

LREFLanglois MR, Blaton VH. Historical milestones in measurement of HDL-cholesterol: Impact on clinical and laboratory practice. Clin Chimica Acta 2006;369:168-178.
Of these, the direct methods are used routinely. Roche HDLC4 is also a direct method. The automated HDLC4 assay uses detergents, cholesterol esterase (CHER), cholesterol oxidase (CHOD) and peroxidase to form a colored pigment that is measured optically.
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

The HDLC4 assay meets the 1998 National Institutes of Health (NIH) / National Cholesterol Education Program (NCEP) goals for precision and accuracy.

LREFKimberly M, Leary E, Cole T, et al. Selection, Validation, Standardization and Performance of a Designated Comparison Method for HDL-Cholesterol for Use in the Cholesterol Reference Method Laboratory Network. Clin Chem 1999;45:1803-1812.
,
LREFSaraf S, Ray KK. Guidelines in the USA, a viewpoint contrary to those guidelines in Europe, Canada, Britain and the International Atherosclerosis Society. Curr Opin Lipidol 2014 Dec;25(6):413-7.

", "Language": "en" }, { "Name": "Reagents", "Value": "

Reagents - working solutions

R1

TAPSOb) buffer: 62.1 mmol/L, pH 7.77; polyanion: 1.25 g/L; EMSE: 1.08 mmol/L; ascorbate oxidase (cucurbita): ≥ 50 μkat/L; peroxidase (horseradish): ≥ 166.7 μkat/L; detergent; BSA: 2.0 g/L; preservative

SR

Bis-Trisc) buffer: 20.1 mmol/L, pH 6.70; cholesterol esterase (microorganism): ≥ 7.5 μkat/L; cholesterol oxidase (recombinant E. coli): ≥ 7.17 μkat/L; cholesterol oxidase (microorganism): ≥ 76.7 μkat/L; peroxidase (horseradish): ≥ 333 μkat/L; 4‑amino‑antipyrine: 1.48 mmol/L; BSA: 3.0 g/L; detergents; preservative

b) 2‑Hydroxy‑N‑tris(hydroxymethyl)methyl‑3‑aminopropanesulfonic acid

c) Bis(2‑hydroxyethyl)iminotris(hydroxymethyl)methane

R1 is in position B and SR is in position C.

", "Language": "en" }, { "Name": "PrecautionsWarnings", "Value": "

Precautions and warnings

For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.

Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.

Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.

Safety data sheet available for professional user on request.

", "Language": "en" }, { "Name": "Caution", "Value": "", "Language": "en" }, { "Name": "QualityControl", "Value": "

Quality control

Reference range

PreciControl ClinChem Multi 2

Pathological range

PreciControl ClinChem Multi 1

Control interval

24 hours recommended

Control sequence

User defined

Control after calibration

Recommended

For quality control, use control materials as listed in the \"Order information\" section.

In addition, other suitable control material can be used.

The control intervals and limits should be adapted to each laboratory’s individual requirements. Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.

Quality control materials are intended for use only as monitors of accuracy and precision.

Follow the applicable government regulations and local guidelines for quality control.

", "Language": "en" }, { "Name": "SpecimenPreparation", "Value": "

Specimen collection and preparation

For specimen collection and preparation only use suitable tubes or collection containers.

Only the specimens listed below were tested and found acceptable.
Serum.
Plasma: Li‑heparin, K2‑ and K3‑EDTA plasma.

The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.

Centrifuge samples containing precipitates before performing the assay.

Collect blood by using an evacuated tube or syringe. Specimens should preferably be analyzed on the day of collection.

Fasting and non‑fasting samples can be used.

LREFSidhu D, Naugler C. Fasting time and lipid levels in a community-based population: A cross sectional study. Arch. Intern. Med. Dec 10, 2012; 172(22):1707-10.
,
LREFOntario Community Laboratory Guideline for Adult Lipid Testing (CLP017) 2013.

Stability in serum:

72 hours at 15‑25 °C

LREFData on file at Roche Diagnostics.

7 days at 2‑8 °C

LREFData on file at Roche Diagnostics.

12 months at -20 °C

LREFJansen EHLM, Beekhof PK, Schenk E. Long Term Stability of Lipid Metabolism in Frozen Human Serum: Triglycerides, Free Fatty Acids, Total-, HDL- and LDL-cholesterol, Apolipoprotein-A1 and B. J Mol Biomark Diagn 2014;5:4.

24 months at -70 °C

LREFShih WJ, Bachorik PS, Haga JA, et al. Estimating the Long-Term Effects of Storage at -70°C on Cholesterol, Triglyceride, and HDL-Cholesterol Measurements in Stored Sera. Clin Chem 2000 Mar;46(3):351-64.

Stability in Li‑heparin, K2‑ and
K3‑EDTA plasma:

72 hours at 15‑25 °C

LREFData on file at Roche Diagnostics.

7 days at 2‑8 °C

LREFData on file at Roche Diagnostics.

3 months at (-15)‑(-25) °C

LREFData on file at Roche Diagnostics.

18 months at -70 °C

LREFData on file at Roche Diagnostics.

18 months at -80 °C

LREFKronenberg F, Lobentanz EM, König P, et al. Effect of sample storage on the measurement of lipoprotein[a], apolipoproteins B and A-IV, total and high density lipoprotein cholesterol and triglycerides. J Lipid Res. 1994 Jul;35(7):1318-28.

It is reported that EDTA stabilizes lipoproteins.

LREFCooper GR, Myers GL, Smith SJ, et al. Standardization of Lipid, Lipoprotein, and Apolipoprotein Measurements. Clin Chem 1988;34(8B):B95-B105.

", "Language": "en" } ] } }, { "ProductSpecVariant": { "MetaData": { "DocumentMaterialNumber": "0107528566190c501", "ProductName": "HDLC4", "ProductLongName": "HDL-Cholesterol Gen.4", "Language": "en", "DocumentVersion": "4", "DocumentObjectID": "FF0000000477150E", "DocumentOriginID": "FF00000001BF620E", "MaterialNumbers": [ "07528566190" ], "InstrumentReferences": [ { "ID": "308", "BrandName": "cobas c 311" }, { "ID": "2324", "BrandName": "cobas c 502" }, { "ID": "309", "BrandName": "cobas c 501" } ], "DisclaimerText": "Product information shown on this page contains elements of the officially released Method Sheet. If you require further information please refer to the full Method Sheet PDF under the given link, or contact your local Roche country representative." }, "Chapters": [ { "Name": "IntendedUse", "Value": "

Intended use

In vitro diagnostic test for the quantitative determination of the HDL‑cholesterol concentration in human serum and plasma on Roche/Hitachi cobas c systems.

", "Language": "en" }, { "Name": "TestPrinciple", "Value": "

Test principle

Test principle
LREFMiida T, Nishimura K, Okamura T, et al. Validation of homogeneous assays for HDL-cholesterol using fresh samples from healthy and diseased subjects. Atherosclerosis 2014;233(1):253-9.
,
LREFKatayama Y, Soya H, Fujinaka M, et al. Evaluation of New Homogeneous Assay Kit to Determine HDL-C with a High Reactivity with Cholesterol in Various Types of HDL. AACC Meeting 2009, Poster Abstract B-103.

Homogeneous enzymatic colorimetric test.

Non‑HDL lipoproteins such as LDL, VLDL and chylomicrons are combined with polyanions and a detergent forming a water‑soluble complex. In this complex the enzymatic reaction of CHER and CHOD towards non‑HDL lipoproteins is blocked.
Finally only HDL‑particles can react with CHER and CHOD. The concentration of HDL‑cholesterol is determined enzymatically by CHER and CHOD.
Cholesterol esters are broken down quantitatively into free cholesterol and fatty acids by CHER.

CHER

HDL‑cholesterol esters + H2O

HDL‑cholesterol + RCOOH

In the presence of oxygen, cholesterol is oxidized by cholesterol oxidase to Δ4‑cholestenone and hydrogen peroxide.

CHOD

HDL‑cholesterol + O2

Δ4‑cholestenone + H2O2

In the presence of peroxidase, the hydrogen peroxide generated reacts with 4‑amino‑antipyrine and EMSEa) to form a dye. The color intensity of this dye is directly proportional to the cholesterol concentration and is measured photometrically.

Peroxidase

2 H2O2 + 4‑amino‑antipyrine +
EMSE + H+ + H2O

colored pigment + 5 H2O

a) N‑ethyl‑N‑(3‑methylphenyl)‑N’‑succinylethylenediamine

", "Language": "en" }, { "Name": "MeasuringRange", "Value": "

Limits and ranges

Measuring range

0.08‑3.88 mmol/L (3.09‑150 mg/dL)

Determine samples having higher concentrations via the rerun function. Dilution of samples via the rerun function is a 1:2 dilution. Results from samples diluted using the rerun function are automatically multiplied by a factor of 2.

Lower limits of measurement

Limit of Blank, Limit of Detection and Limit of Quantitation

Limit of Blank

= 0.08 mmol/L (3.09 mg/dL)

Limit of Detection

= 0.08 mmol/L (3.09 mg/dL)

Limit of Quantitation

= 0.08 mmol/L (3.09 mg/dL)

The Limit of Blank, Limit of Detection and Limit of Quantitation were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A2 requirements.

The Limit of Blank is the 95th percentile value from n ≥ 60 measurements of analyte‑free samples over several independent series. The Limit of Blank corresponds to the concentration below which analyte‑free samples are found with a probability of 95 %.

The Limit of Detection is determined based on the Limit of Blank and the standard deviation of low concentration samples.

The Limit of Detection corresponds to the lowest analyte concentration which can be detected (value above the Limit of Blank with a probability of 95 %).

The Limit of Quantitation is the lowest analyte concentration that can be reproducibly measured with a precision of ≤ 30 % CV. It has been determined using low concentration HDL‑cholesterol samples.

", "Language": "en" }, { "Name": "ExpectedValues", "Value": "

Expected values

No risk

Moderate risk

High risk

Females

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.68 mmol/L

1.15‑1.68 mmol/L

< 1.15 mmol/L

(> 65 mg/dL)

(45‑65 mg/dL)

(< 45 mg/dL)

Males

LREFThomas L, ed. Labor und Diagnose, 4th ed. Marburg: Die Medizinische Verlagsgesellschaft 1992;208.
,
LREFAssmann G. At what levels of total low- or high-density lipoprotein cholesterol should diet/drug therapy be initiated? European guidelines. Amer J Cardiol 1990;65:11F.
,
LREFAssmann G, Schriewer H, Schmitz G, et al.Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2. Clin Chem 1983;29(12):2026-2030.

> 1.45 mmol/L

0.90‑1.45 mmol/L

< 0.90 mmol/L

(> 55 mg/dL)

(35‑55 mg/dL)

(< 35 mg/dL)

National Cholesterol Education Program (NCEP) guidelines:

LREFThird Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III). NIH Publication No 01-3670; May 2001.

< 40 mg/dL: Low HDL‑cholesterol (major risk factor for CHD)

≥ 60 mg/dL: High HDL‑cholesterol (“negative” risk factor for CHD)

HDL‑cholesterol is affected by a number of factors, e.g. smoking, exercise, hormones, sex and age.

Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.

National Cholesterol Education Program (NCEP) guidelines are based on serum values. When classifying patients, serum or serum equivalent values should be used. Therefore the NCEP recommends using a factor of 1.03 to convert EDTA plasma values to serum values. A later study found EDTA plasma concentrations to be 4.7 % lower than those in serum.

LREFCloey T, Bachorik PS, Becker D, et al. Reevaluation of Serum-Plasma Differences in Total Cholesterol Concentration. JAMA 1990 May 23-30;263(20):2788-9.
To comply with the 1998 NCEP goal of a bias < 5 % it is recommended that each laboratory validates this conversion factor and enters it into the test parameters for HDL‑cholesterol.
LREFNational Cholesterol Education Program Recommendations for Measurement of High-Density Lipoprotein Cholesterol: Executive Summary. Clin Chem 1995;41:1427-1433.

Treatment goals for non‑HDL‑cholesterol have been proposed:

LREFBlaha MJ, Blumenthal RS, Brinton EA, et al. The importance of non-HDL cholesterol reporting in lipid management. J Clin Lipidol 2008 Aug;2(4):267-73.

NCEP ATP III

ADA/AHA Guidelines for patients with increased cardiometabolic risk

Optional goal for very-high/highest risk patients (known CVD, diabetes with elevated risk)

< 3.37 mmol/L
(< 130 mg/dL)

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for those with established cardiovascular disease and multiple major risk factors

< 2.59 mmol/L
(< 100 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for moderately-high/intermediate risk patients (≥ 2 major CVD risk factors, Framingham 10‑year risk score from 10‑20 %)

< 4.14 mmol/L
(< 160 mg/dL)

< 3.37 mmol/L
(< 130 mg/dL)

Optional goal for high-risk patients, CHD-risk-equivalent (Framingham 10‑year risk score > 20 %/10 years, diabetes without other major risk factors)

< 3.37 mmol/L
(< 130 mg/dL)

", "Language": "en" }, { "Name": "LimitationInterference", "Value": "

Limitations - interference

Limitations - interference
LREFKadri N, Douville P, Lachance P. Letter to editor. Clin Chem 2002;48:964.

Criterion: Recovery within ± 10 % of initial value at a HDL‑cholesterol concentration of 1 mmol/L (38.7 mg/dL).

Icterus:

LREFGlick MR, Ryder KW, Jackson SA. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470-475.
No significant interference up to an I index of 60 for conjugated and unconjugated bilirubin (appr