In vitro test for the quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in whole blood and hemolysates prepared from whole blood on the cobas c 111 system. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus. Moreover, this test is to be used as an aid in diagnosis of diabetes and identifying patients who may be at risk for developing diabetes.
This method uses TTAB (Tetradecyltrimethylammonium bromide) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). Sample pretreatment to remove labile HbA1c is not necessary.
All hemoglobin variants which are glycated at the β‑chain N‑terminus and which have antibody-recognizable regions identical to that of HbA1c are measured by this assay. Consequently, the metabolic state of patients having uremia or the most frequent hemoglobinopathies (HbAS, HbAC, HbAE) can be determined using this assay.
Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
Sample and addition of R1 (buffer/antibody):
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, complex formation does not take place.
Addition of SR (buffer/polyhapten) and start of reaction:
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibody-polyhapten complex which can be measured turbidimetrically.
Hemoglobin
Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently not necessary.
The final result is expressed as mmol/mol HbA1c or percent HbA1c and is calculated from the HbA1c/Hb ratio as follows:
Protocol 1 (mmol/mol HbA1c acc. to IFCC):
HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
Protocol 2 (% HbA1c acc. to DCCT/NGSP):
HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
Hb: 2.48‑24.8 mmol/L (4‑40 g/dL)
HbA1c: 0.186‑1.61 mmol/L (0.3‑2.6 g/dL)*
*The measuring range for HbA1c lies between 0.186 mmol/L and the concentration of the highest standard. The test range stated above is based on a typical calibrator value of 1.61 mmol/L.
This corresponds to a measuring range of 23‑196 mmol/mol HbA1c (IFCC) and 4.2‑20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L (13.2 g/dL).
In rare cases of \">test rng\" flags that might occur with the whole blood application remix the whole blood sample and repeat the analysis with the same settings.
Hb: | Limit of Blank | = 0.31 mmol/L (0.50 g/dL) |
Limit of Detection | = 0.62 mmol/L (1.00 g/dL) | |
HbA1c: | Limit of Blank | = 0.12 mmol/L (0.19 g/dL) |
Limit of Detection | = 0.18 mmol/L (0.29 g/dL) |
The Limit of Blank and Limit of Detection were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A 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 %).
Protocol 1 (mmol/mol HbA1c acc. to IFCC): 29‑42 mmol/mol HbA1c
Protocol 2 (% HbA1c acc. to DCCT/NGSP): 4.8‑5.9 % HbA1c
This reference range was obtained by measuring
HbA1c levels may reach 195 mmol/mol HbA1c (IFCC) or 20 % HbA1c (DCCT/NGSP) and more in poorly controlled diabetes. Therapeutic action is suggested at levels above 64 mmol/mol HbA1c (IFCC) or 8 % HbA1c (DCCT/NGSP). Diabetes patients with HbA1c levels below 53 mmol/mol HbA1c (IFCC) or 7 % HbA1c (DCCT/NGSP) meet the goal of the American Diabetes Association.
HbA1c levels below the established reference range may indicate recent episodes of hypoglycemia, the presence of Hb variants, or shortened lifetime of erythrocytes.
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
For diagnostic purposes, mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) should be used in conjunction with information from other diagnostic procedures and clinical evaluations.
The test is designed only for accurate and precise measurement of mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP). The individual results for total Hb and HbA1c concentration should not be reported.
As a matter of principle, care must be taken when interpreting any HbA1c result from patients with Hb variants. Abnormal hemoglobins might affect the half life of the red cells or the in vivo glycation rates. In these cases even analytically correct results do not reflect the same level of glycemic control that would be expected in patients with normal hemoglobin.
Any cause of shortened erythrocyte survival will reduce exposure of erythrocytes to glucose with a consequent decrease in mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP), even though the time-averaged blood glucose level may be elevated. Causes of shortened erythrocyte lifetime might be hemolytic anemia or other hemolytic diseases, homozygous sickle cell trait, pregnancy, recent significant or chronic blood loss, etc. Caution should be used when interpreting the HbA1c results from patients with these conditions. HbA1c must not be used for the diagnosis of diabetes mellitus in the presence of such conditions.
Glycated HbF is not detected by the assay as it does not contain the glycated β-chain that characterizes HbA1c. However, HbF is measured in the total Hb assay and as a consequence, specimens containing high amounts of HbF (≥ 10 %) may result in lower than expected mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).
mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) are not suitable for diagnosis of gestational diabetes.
In very rare cases of rapidly evolving type 1 diabetes the increase of HbA1c values might be delayed compared to the acute increase in glucose concentrations. In these conditions diabetes mellitus must be diagnosed based on plasma glucose concentrations and/or the typical clinical symptoms.
Criterion: Recovery within ± 10 % of initial value.
Icterus: No significant interference up to a conjugated and unconjugated bilirubin concentration of 1026 μmol/L or 60 mg/dL.
Lipemia: No significant interference up to an Intralipid concentration of 600 mg/dL. There is poor correlation between the triglycerides concentration and turbidity.
Glycemia: No significant interference up to a glucose level of 55.5 mmol/L (1000 mg/dL). A fasting sample is not required.
Rheumatoid factors: No significant interference up to a rheumatoid factor level of 750 IU/mL.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated hemoglobin, carbamylated hemoglobin, glycated albumin and labile HbA1c were found for the anti-HbA1c antibodies used in this kit.
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.
|
| Analyzer(s) on which kit(s) can be used | |
05336180 190 | Tina-quant Hemoglobin A1c Gen.3 (2 × 100 tests) | cobas c 111 |
Materials required (but not provided):
04528417 190 | C.f.a.s. HbA1c (3 × 2 mL) | Code 674 | |
05479207 190 | PreciControl HbA1c norm (4 × 1 mL) | Code 208 | |
05912504 190 | PreciControl HbA1c path (4 × 1 mL) | Code 209 | |
05007232 190 | Hemolyzing Reagent Gen.2 (8 × 6.3 mL) | Code 952 | |
11488457 122 | HbA1c Hemolyzing Reagent for Tina-quant HbA1c (1000 mL) | ||
11930630 001 | Chimneys |
Whole Blood DCCT (%): ACN 891 A1W3D and ACN 871 HBW3D
Hemolysate DCCT (%): ACN 861 A1H3D and ACN 841 HBH3D
Whole Blood IFCC (mmol/mol): ACN 264 A1W3I and ACN 265 HBW3I
Hemolysate IFCC (mmol/mol): ACN 251 A1H3I and ACN 231 HBH3I
Ready for use
Under conditions of extreme humidity, condensation may lead to a dilution of the reagent that affects the measurements. Hence under environmental conditions in which temperature and humidity are equal to, or in excess of 25 °C/80 %, 28 °C/70 %, 30 °C/60 % or 32 °C/55 % a chimney (Cat. No. 11930630 001) should be used to reduce the condensation rate. Place a white chimney in R1 and a black chimney in SR. The chimneys can be reused for reagent bottles within the same kit. However, to avoid contamination of the reagent with detergent or dilution of the reagent with water it is not permitted to wash the chimneys before reuse.
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction direction | Increase |
Wavelength A/B | 378/659 nm |
Calc. first/last | 6/16 |
Unit | mmol/L |
Reaction mode | R1-S |
Hb | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
Total volume | 126 µL |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction direction | Increase |
Wavelength A/B | 340/659 nm |
Calc. first/last | 16/34 |
Unit | mmol/L |
Reaction mode | R1-S-SR |
HbA1c | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
SR | 24 µL | 0 µL |
Total volume | 150 µL |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction direction | Increase |
Wavelength A/B | 378/659 nm |
Calc. first/last | 6/16 |
Unit | mmol/L |
Reaction mode | R1-S |
Hb | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
Total volume | 126 µL |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction direction | Increase |
Wavelength A/B | 340/659 nm |
Calc. first/last | 16/34 |
Unit | mmol/L |
Reaction mode | R1-S-SR |
HbA1c | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
SR | 24 µL | 0 µL |
Total volume | 150 µL |
Shelf life at 2‑8 °C: | See expiration date on reagent |
On-board in use and refrigerated on the analyzer: | 4 weeks |
Shelf life at 2‑8 °C: | See expiration date on vial label |
On-board in use and refrigerated on the analyzer: | 5 days |
When storing at temperatures under 3 °C, the reagent may become cloudy. This has no effect on the function of the reagent and is reversible at higher temperatures. It is therefore recommended to equilibrate the reagent at room temperature for approximately 10 minutes and mix thoroughly before use.
Calibrator | C.f.a.s. HbA1c |
Calibrator | C.f.a.s. HbA1c |
Calibration dilution ratio | 1:1, 1:1.5, 1:2.1, 1:3, 1:6, 1:15, performed automatically by the instrument |
Calibrator diluent | Hemolyzing Reagent Gen.2 |
Calibration frequency | Each lot, every 14 days, 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 approved IFCC reference method for the measurement of HbA1c in human blood
Enter the assigned lot-specific and application-specific value of the calibrator. Use the appropriate C.f.a.s. HbA1c calibrator only.
The cobas c 111 Hemolyzing Reagent Gen.2 (Cat. No. 05007232 190) needs to be available on the analyzer. Otherwise the calibration cannot be performed.
Representative performance data on the cobas c 111 analyzer are given below. Results obtained in individual laboratories may differ.
Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained (data based on DCCT/NGSP values):
Repeatability | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.63 | 0.04 | 0.7 |
PreciControl HbA1c path | 10.7 | 0.06 | 0.6 |
Human sample 1 | 5.21 | 0.04 | 0.7 |
Human sample 2 | 6.22 | 0.04 | 0.7 |
Human sample 3 | 7.34 | 0.05 | 0.7 |
Human sample 4 | 8.44 | 0.05 | 0.6 |
Human sample 5 | 11.0 | 0.06 | 0.5 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.63 | 0.08 | 1.5 |
PreciControl HbA1c path | 10.7 | 0.10 | 0.9 |
Human sample 1 | 5.21 | 0.07 | 1.4 |
Human sample 2 | 6.22 | 0.07 | 1.1 |
Human sample 3 | 7.34 | 0.09 | 1.2 |
Human sample 4 | 8.44 | 0.09 | 1.1 |
Human sample 5 | 11.0 | 0.10 | 0.9 |
Repeatability | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.76 | 0.06 | 1.0 |
PreciControl HbA1c path | 10.5 | 0.06 | 0.6 |
Human sample 1 | 4.99 | 0.07 | 1.5 |
Human sample 2 | 6.42 | 0.08 | 1.3 |
Human sample 3 | 10.2 | 0.07 | 0.7 |
Human sample 4 | 7.44 | 0.07 | 0.9 |
Human sample 5 | 7.69 | 0.05 | 0.7 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.76 | 0.12 | 2.1 |
PreciControl HbA1c path | 10.5 | 0.19 | 1.8 |
Human sample 1 | 4.99 | 0.11 | 2.2 |
Human sample 2 | 6.42 | 0.16 | 2.5 |
Human sample 3 | 10.2 | 0.18 | 1.8 |
Human sample 4 | 7.44 | 0.14 | 1.9 |
Human sample 5 | 7.69 | 0.13 | 1.6 |
Evaluation of method comparison data is according to former NGSP certification criteria. The mean difference between the two methods and the 95 % confidence intervals of the differences in the range from 4‑10 % (DCCT/NGSP) are given. 95 % of the differences between the values obtained for individual samples with both methods fall within the range defined by the lower and upper 95 % confidence intervals of the differences.
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 57 |
Mean difference: 0.22 % HbA1c |
Lower 95 % confidence interval of differences: −0.22 % HbA1c |
Upper 95 % confidence interval of differences: 0.66 % HbA1c |
The sample concentrations were between 4.86 % and 12.2 % HbA1c (DCCT/NGSP values).
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 57 |
Mean difference: 0.19 % HbA1c |
Lower 95 % confidence interval of differences: −0.16 % HbA1c |
Upper 95 % confidence interval of differences: 0.54 % HbA1c |
The sample concentrations were between 4.58 % and 11.8 % HbA1c (DCCT/NGSP values).
Hemoglobin (Hb) consists of four protein subunits, each containing a heme moiety, and is the red-pigmented protein located in the erythrocytes. Its main function is to transport oxygen and carbon dioxide in blood. Each Hb molecule is able to bind four oxygen molecules. Hb consists of a variety of subfractions and derivatives. Among this heterogeneous group of hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction formed by the attachment of various sugars to the Hb molecule. HbA1c is formed in two steps by the nonenzymatic reaction of glucose with the N‑terminal amino group of the β‑chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This is rearranged to form stable HbA1c in a second reaction step.
In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte’s life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. Glucose levels closer to the time of the assay have a greater influence on the HbA1c level.
The approximate relationship between HbA1c and mean blood glucose values during the preceding 2 to 3 months was analyzed in several studies. A recent study obtained the following correlation:
IFCC standardization (recalculated acc. to ref. 8)
• Estimated average glucose [mmol/L] = 0.146 × HbA1c (mmol/mol) + 0.834
or
• Estimated average glucose [mg/dL] = 2.64 × HbA1c (mmol/mol) + 15.03
Standardization acc. to DCCT/NGSP
• Estimated average glucose [mmol/L] = 1.59 × HbA1c (%) - 2.59
or
• Estimated average glucose [mg/dL] = 28.7 × HbA1c (%) - 46.7
The risk of diabetic complications, such as diabetic nephropathy and retinopathy, increases with poor metabolic control. In accordance with its function as an indicator for the mean blood glucose level, HbA1c predicts the development of diabetic complications in diabetes patients.
For monitoring of long term glycemic control, testing every 3 to 4 months is generally sufficient. In certain clinical situations, such as gestational diabetes, or after a major change in therapy, it may be useful to measure HbA1c in 2 to 4 week intervals.
R1 | Antibody reagent FREFMES = 2-morpholinoethane sulfonic acid buffer: 0.025 mol/L; TRISFREFTRIS = Tris(hydroxymethyl)-aminomethane buffer: 0.015 mol/L, pH 6.2; HbA1c antibody (ovine serum) ≥ 0.5 mg/mL; detergents; stabilizers; preservative |
SR | Polyhapten reagent |
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.
This kit contains components classified as follows in accordance with the Regulation (EC) No. 1272/2008:
|
Warning
H317 | May cause an allergic skin reaction. |
P261 | Avoid breathing dust/fume/gas/mist/vapours/spray. |
P272 | Contaminated work clothing should not be allowed out of the workplace. |
P280 | Wear protective gloves. |
P333 + P313 | If skin irritation or rash occurs: Get medical advice/attention. |
P362 + P364 | Take off contaminated clothing and wash it before reuse. |
P501 | Dispose of contents/container to an approved waste disposal plant. |
Product safety labeling follows EU GHS guidance.
Contact phone: all countries: +49-621-7590
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.
Follow the applicable government regulations and local guidelines for quality control.
HbA1c controls carry a declaration for HbA1c ratio only. No declarations for Hb and HbA1c concentrations are provided. HbA1c controls must be included in the cobas c 111 systems Quality Control Program. For the hemolysate test the HbA1c controls must be hemolysed manually before using them for the Quality Control Program. Assigned target values in mmol/mol HbA1c according to IFCC are not included in the control barcodes and must be entered by the customer manually.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Anticoagulated venous or capillary blood or hemolysate
The only acceptable anticoagulants are Li‑Heparin, K2‑EDTA, K3‑EDTA, Fluoride/Na2‑EDTA, Na‑Heparin and Fluoride/potassium oxalate.
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.
See the limitations and interferences section for details about possible sample interferences.
Stability: | 3 days at 15‑25 °C |
The recovery of HbA1c ratio values from sedimented samples, especially in case of poorly controlled diabetic patients, may be slightly elevated. To minimize this effect samples may be gently mixed by inversion prior to analysis.
Freeze only once. Mix specimen thoroughly after thawing.
Whole blood samples must be placed on the instrument in uncapped primary tubes and not in (micro) tubes.
Manual hemolysate preparation:
Allow blood specimen and Hemolyzing Reagent for Tina‑quant HbA1c (Cat. No. 11488457122) to equilibrate at room temperature before use.
Moderately mix the sample immediately prior to pipetting, to ensure homogeneous mixture of erythrocytes. Take care to avoid the formation of foam.
Dilute the sample with Hemolyzing Reagent for Tina‑quant HbA1c in the ratio 1:101 (1+100) using one of the following pipetting schemes.
Pipette into tubes:
Hemolyzing Reagent for Tina-quant HbA1c: 500 µL
Specimen (patient or control): 5 µL
or
Hemolyzing Reagent for Tina-quant HbA1c: 1000 µL
Specimen (patient or control): 10 µL
or
Hemolyzing Reagent for Tina-quant HbA1c: 2000 µL
Specimen (patient or control): 20 µL
Mix using a vibration mixer or by gentle swirling.
The hemolysate can be used after the solution has changed color from red to brownish-green (approximately 1‑2 min).
Stability of the hemolysate: | 4 hours at 15‑25 °C |
In vitro test for the quantitative determination of mmol/mol hemoglobin (IFCC) and % hemoglobin A1c (DCCT/NGSP) in whole blood or hemolysate on Roche/Hitachi cobas c systems. HbA1c determinations are useful monitoring of long-term blood glucose control in individuals with diabetes mellitus. Moreover, this test is to be used as an aid in diagnosis of diabetes and identifying patients who may be at risk for developing diabetes.
This method uses TTABa) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). Sample pretreatment to remove labile HbA1c is not necessary.
All hemoglobin variants which are glycated at the β‑chain N‑terminus and which have antibody-recognizable regions identical to that of HbA1c are determined by this assay. Consequently, the metabolic state of patients having uremia or the most frequent hemoglobinopathies (HbAS, HbAC,
a) TTAB = Tetradecyltrimethylammonium bromide
Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
Sample and addition of R1 (buffer/antibody)
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, formation of insoluble complexes does not take place.
Addition of R3 (buffer/polyhapten) and start of reaction:
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibody-polyhapten complex which can be determined turbidimetrically.
Hemoglobin
Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently not necessary.
The final result is expressed as mmol/mol HbA1c or % HbA1c and is calculated from the HbA1c/Hb ratio as follows:
Protocol 1 (mmol/mol HbA1c acc. to IFCC):
HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
Protocol 2 (% HbA1c acc. to DCCT/NGSP):
HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
Measuring range
Hemoglobin: 2.48‑24.8 mmol/L (4‑40 g/dL).
HbA1c: 0.186‑1.61 mmol/L (0.3‑2.6 g/dL)
This corresponds to a measuring range of 23‑196 mmol/mol HbA1c (IFCC) and 4.2‑20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L (13.2 g/dL).
In rare cases of “>Test” flags which might occur with the use of the whole blood application, remix the whole blood sample and repeat the analysis with the same settings.
It is recommended to switch the auto rerun function off.
Lower limits of measurement
Limit of Blank and Limit of Detection
Hemoglobin:
Limit of Blank = 0.31 mmol/L (0.50 g/dL)
Limit of Detection = 0.62 mmol/L (1.00 g/dL)
HbA1c:
Limit of Blank = 0.12 mmol/L (0.19 g/dL)
Limit of Detection = 0.18 mmol/L (0.29 g/dL)
The Limit of Blank and Limit of Detection were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A 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 sample concentration which leads with a probability of 95 % to a measurement result above the Limit of Blank.
Protocol 1 (mmol/mol HbA1c acc. to IFCC): 29‑42 mmol/mol HbA1c
Protocol 2 (% HbA1c acc. to DCCT/NGSP): 4.8‑5.9 % HbA1c
This reference range was obtained by measuring
HbA1c levels may reach 195 mmol/mol (IFCC) or 20 % (DCCT/NGSP) or higher in poorly controlled diabetes. Therapeutic action is suggested at levels above 64 mmol/mol HbA1c (IFCC) or 8 % HbA1c (DCCT/NGSP). Diabetes patients with HbA1c levels below 53 mmol/mol (IFCC) or 7 % (DCCT/NGSP) meet the goal of the American Diabetes Association.
HbA1c levels below the established reference range may indicate recent episodes of hypoglycemia, the presence of Hb variants, or shortened lifetime of erythrocytes.
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
1. For diagnostic purposes, mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) should be used in conjunction with information from other diagnostic procedures and clinical evaluations.
2. The test is designed only for accurate and precise measurement of mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP). The individual results for total Hb and HbA1c concentration should not be reported.
3. As a matter of principle, care must be taken when interpreting any HbA1c result from patients with Hb variants. Abnormal hemoglobins might affect the half life of the red cells or the in vivo glycation rates. In these cases even analytically correct results do not reflect the same level of glycemic control that would be expected in patients with normal hemoglobin.
4. Any cause of shortened erythrocyte survival or decrease in mean erythrocyte age will reduce exposure of erythrocytes to glucose with a consequent decrease in mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP), even though the time-averaged blood glucose level may be elevated. Causes of shortened erythrocyte lifetime might be hemolytic anemia or other hemolytic diseases, homozygous sickle cell trait, pregnancy, recent significant or chronic blood loss, etc. Similarly, recent blood transfusions can alter the mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).Caution should be used when interpreting the HbA1c results from patients with these conditions. HbA1c must not be used for the diagnosis of diabetes mellitus in the presence of such conditions.
5. Glycated HbF is not detected by the assay as it does not contain the glycated β‑chain that characterizes HbA1c. However, HbF is measured in the total Hb assay and as a consequence, specimens containing high amounts of HbF (> 10 %) may result in lower than expected mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).
6. mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) are not suitable for the diagnosis of gestational diabetes.
7. In very rare cases of rapidly evolving type 1 diabetes the increase of the HbA1c values might be delayed compared to the acute increase in glucose concentrations. In these conditions diabetes mellitus must be diagnosed based on plasma glucose concentrations and/or the typical clinical symptoms.
Criterion: Recovery within ± 10 % of initial value.
Icterus:
Lipemia (Intralipid):
Glycemia: No significant interference up to a glucose level of 55.5 mmol/L (1000 mg/dL). A fasting sample is not required.
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 750 IU/mL.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated hemoglobin, carbamylated hemoglobin, glycated albumin and labile HbA1c were found for the anti-HbA1c antibodies used in this kit.
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. 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.
| Analyzer(s) on which cobas c pack(s) can be used | ||
05336163 190 | Tina-quant Hemoglobin A1c Gen.3 (150 tests) | System ID 07 7455 3 | cobas c 311, cobas c 501/502 |
Materials required (but not provided):
04528417 190 | Calibrator f.a.s. HbA1c (3 x 2 mL) | Code 674 | |
05479207 190 | PreciControl HbA1c norm (4 × 1 mL) | Code 208 | |
05912504 190 | PreciControl HbA1c path (4 × 1 mL) | Code 209 | |
04528182 190 | Hemolyzing Reagent Gen.2 (51 mL)* | System ID 07 6873 1 | |
11488457 122 | HbA1c Hemolyzing Reagent for Tina‑quant HbA1c (1000 mL) | For Hemolysate Application only |
* The value encoded in the instrument settings is 45 mL to account for the dead volume of the bottles.
Whole Blood Application - Standardized according to IFCC transferable to DCCT/NGSP | ||
HB-W3: | ACN 871 | Hemoglobin (Hb) |
A1-W3: | ACN 881 | Hemoglobin A1c (HbA1c) |
RWD3: | ACN 891 | Ratio % HbA1c (acc. to DCCT/NGSP) |
A1CD2: | ACN 952 | Hemolyzing reagent |
Hemolysate Application - Standardized according to IFCC transferable to DCCT/NGSP | ||
HB-H3: | ACN 841 | Hemoglobin (Hb) |
A1–H3: | ACN 851 | Hemoglobin A1c (HbA1c) |
RHD3: | ACN 861 | Ratio % HbA1c (acc. to DCCT/NGSP) |
A1CD2: | ACN 952 | Hemolyzing reagent |
Ready for use
Whole Blood application for Hb (HB‑W3) and HbA1c (A1‑W3)
cobas c 311 test definition Hb (HB-W3) | |||
Assay type | 1-Point | ||
Reaction time / Assay points | 10 / 23 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
cobas c 311 test definition HbA1c (A1-W3) | |||
Assay type | 2-Point End | ||
Reaction time / Assay points | 10 / 23‑57 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
cobas c 501/502 test definition Hb (HB-W3) | |||
Assay type | 1-Point | ||
Reaction time / Assay points | 10 / 34 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
cobas c 501/502 test definition HbA1c (A1-W3) | |||
Assay type | 2-Point End | ||
Reaction time / Assay points | 10 / 34-70 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
Ratio definition for mmol/mol HbA1c and % HbA1c calculation | |
Abbreviated ratio name | RWI3 |
Equation | (A1-W3/HB-W3) × 1000 |
Unit | mmol/mol |
Protocol 2 (% HbA1c acc. to DCCT/NGSP): | |
Abbreviated ratio name | RWD3 (891) |
Equation | (A1-W3/HB-W3) × 91.5 + 2.15 |
Unit | % |
Protocol 2 is already implemented in the application (ACN 891). The mmol/mol HbA1c values according to Protocol 1 (IFCC) must be manually calculated according to the above equation. If requested a calculated test with the formula in protocol 1 can be programmed under Utility > calculated test on the
Sample Type | Supernt. |
Unit of Measure | mM/M |
Report Name | HbA1c Gen.3 IFCC |
Item | RWI3 |
Formula | (A1-W3/HB-W3) x 1000 |
The ratio for HbA1c (mmol/mol HbA1c acc. to IFCC and % HbA1c acc. to DCCT/NGSP) will be automatically calculated after result output of both tests. It is recommended to report % HbA1c values (DCCT/NGSP) to one decimal place and mmol/mol HbA1c values (IFCC) without decimal places, which can be entered in the editable field “expected values”.
Hemolysate Application for Hb (HB‑H3) and HbA1c (A1‑H3)
cobas c 311 test definition Hb (HB-H3) | |||
Assay type | 1-Point | ||
Reaction time / Assay points | 10 / 23 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
cobas c 311 test definition HbA1c (A1-H3) | |||
Assay type | 2-Point End | ||
Reaction time / Assay points | 10 / 23-57 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
cobas c 501/502 test definition Hb (HB-H3) | |||
Assay type | 1‑Point | ||
Reaction time / Assay points | 10 / 34 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
cobas c 501/502 test definition HbA1c (A1-H3) | |||
Assay type | 2‑Point End | ||
Reaction time / Assay points | 10 / 34-70 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
Ratio definition for HbA1c (mmol/mol (IFCC) or % (DCCT/NGSP)) calculation
Protocol 1 (mmol/mol HbA1c acc. to IFCC): | |
Abbreviated ratio name | RHI3 |
Equation | (A1-H3/HB-H3) x 1000 |
Unit | mmol/mol |
Protocol 2 (% HbA1c acc. to DCCT/NGSP): | |
Abbreviated ratio name | RHD3 (861) |
Equation | (A1-H3/HB-H3) x 91.5 + 2.15 |
Unit | % |
Protocol 2 is already implemented in the application (ACN 861). The mmol/mol HbA1c values according to Protocol 1 (IFCC) must be manually calculated according to the above equation. If requested a calculated test with the formula in protocol 1 can be programmed under Utility > calculated test on the
Sample Type | Supernt. |
Unit of Measure | mM/M |
Report Name | HbA1c Gen.3 IFCC |
Item | RHI3 |
Formula | (A1-H3/HB-H3) x 1000 |
The ratio for HbA1c (mmol/mol HbA1c acc. to IFCC and % HbA1c acc. to DCCT/NGSP) will be automatically calculated after result output of both tests. It is recommended to report % HbA1c values (DCCT/NGSP) to one decimal place and mmol/mol HbA1c values (IFCC) without decimal places, which can be entered in the editable field “expected values”.
A1C-3 | |
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 |
Hemolyzing reagent | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
When storing at temperatures under 3 °C, the reagent may become cloudy. This has no effect on the function of the reagent and is reversible at higher temperatures. It is therefore recommended to equilibrate the reagent at room temperature for approximately 10 minutes and mix thoroughly before use.
On-board in use and refrigerated on the analyzer: | 4 weeks |
Calibration for Whole Blood and Hemolysate Application
Hb | |
Calibrators | S1-S2: C.f.a.s. HbA1c |
Calibration mode | Linear |
HbA1c | |
Calibrators | S1-S6: C.f.a.s. HbA1c |
Calibration mode | Spline |
Calibration frequency | Hb and HbA1c: full calibration is recommended - after 29 days during shelf life - after reagent lot change - as required following quality control procedures |
Always calibrate both assays (Hb and HbA1c) in parallel. Automatic calibration at QC failure should be deactivated. |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the approved IFCC reference method for the measurement of HbA1c in human blood
Note for Whole Blood and Hemolysate Application
Enter the assigned lot-specific and application-specific value of the calibrator. Use the appropriate C.f.a.s. HbA1c calibrator only.
The cobas c Hemolyzing Reagent Gen.2 pack, 51 mL, Cat. No. 04528182 190, needs to be available on the analyzer otherwise the calibration cannot be performed.
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements with repeatability and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained (data based on DCCT/NGSP values):
Whole Blood Application:
Repeatability | Mean % HbA1c | SD % | CV % |
PreciControl HbA1c norm | 5.3 | 0.07 | 1.3 |
PreciControl HbA1c path | 9.9 | 0.11 | 1.1 |
Human sample 1 | 4.4 | 0.07 | 1.6 |
Human sample 2 | 5.6 | 0.09 | 1.6 |
Human sample 3 | 8.0 | 0.08 | 1.0 |
Human sample 4 | 10.6 | 0.11 | 1.1 |
Intermediate precision | Mean % HbA1c | SD % | CV % |
PreciControl HbA1c norm | 5.3 | 0.08 | 1.4 |
PreciControl HbA1c path | 9.9 | 0.15 | 1.5 |
Human sample 1 | 4.4 | 0.09 | 1.9 |
Human sample 2 | 5.6 | 0.11 | 2.0 |
Human sample 3 | 8.0 | 0.11 | 1.4 |
Human sample 4 | 10.6 | 0.16 | 1.5 |
Hemolysate Application:
Repeatability | Mean % HbA1c | SD % | CV % |
PreciControl HbA1c norm | 5.1 | 0.07 | 1.3 |
PreciControl HbA1c path | 10.2 | 0.10 | 1.0 |
Human sample 1 | 4.3 | 0.06 | 1.4 |
Human sample 2 | 5.6 | 0.07 | 1.2 |
Human sample 3 | 8.2 | 0.08 | 1.0 |
Human sample 4 | 10.9 | 0.11 | 1.0 |
Intermediate precision | Mean % HbA1c | SD % | CV % |
PreciControl HbA1c norm | 5.1 | 0.11 | 2.2 |
PreciControl HbA1c path | 10.2 | 0.21 | 2.0 |
Human sample 1 | 4.3 | 0.10 | 2.3 |
Human sample 2 | 5.6 | 0.09 | 1.6 |
Human sample 3 | 8.2 | 0.16 | 1.9 |
Human sample 4 | 10.9 | 0.22 | 2.0 |
Evaluation of method comparison data is according to NGSP certification criteria. The mean difference between the two methods and the 95 % confidence intervals of the differences in the range from 4‑10 % (DCCT/NGSP) are given. 95 % of the differences between the values obtained for individual samples with both methods fall within the range defined by the lower and upper 95 % confidence intervals of the differences.
Whole Blood Application:
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 80 | |
Mean difference | 0.07 % HbA1c |
Lower 95 % confidence interval of differences | -0.27 % HbA1c |
Upper 95 % confidence interval of differences | 0.42 % HbA1c |
The sample concentrations were between 4.7 % and 9.8 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 82 | |
Mean difference | 0.07 % HbA1c |
Lower 95 % confidence interval of differences | -0.50 % HbA1c |
Upper 95 % confidence interval of differences | 0.65 % HbA1c |
The sample concentrations were between 5.0 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 80 | |
Mean difference | -0.09 % HbA1c |
Lower 95 % confidence interval of differences | -0.46 % HbA1c |
Upper 95 % confidence interval of differences | 0.28 % HbA1c |
The sample concentrations were between 4.7 % and 9.8 % (DCCT/NGSP values). |
Hemolysate Application:
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 111 | |
Mean difference | -0.19 % HbA1c |
Lower 95 % confidence interval of differences | -0.52 % HbA1c |
Upper 95 % confidence interval of differences | 0.14 % HbA1c |
The sample concentrations were between 4.6 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 84 | |
Mean difference | -0.06 % HbA1c |
Lower 95 % confidence interval of differences | -0.53 % HbA1c |
Upper 95 % confidence interval of differences | 0.41 % HbA1c |
The sample concentrations were between 5.5 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 111 | |
Mean difference | -0.35 % HbA1c |
Lower 95 % confidence interval of differences | -0.68 % HbA1c |
Upper 95 % confidence interval of differences | -0.02 % HbA1c |
The sample concentrations were between 4.7 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a
Sample size (n) = 113 | |
Mean difference | -0.10 % HbA1c |
Lower 95 % confidence interval of differences | -0.49 % HbA1c |
Upper 95 % confidence interval of differences | 0.31 % HbA1c |
The sample concentrations were between 4.8 % and 9.7 % (DCCT/NGSP values). |
Hemoglobin (Hb) consists of four protein subunits, each containing a heme moiety, and is the red-pigmented protein located in the erythrocytes. Its main function is the transport of oxygen and carbon dioxide in blood. Each Hb molecule is able to bind four oxygen molecules. Hb consists of a variety of subfractions and derivatives. Among this heterogeneous group of hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction formed by the attachment of various sugars to the Hb molecule. HbA1c is formed in two steps by the non-enzymatic reaction of glucose with the N‑terminal amino group of the β‑chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This is rearranged to form stable HbA1c in a second reaction step.
In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte's life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. Glucose levels closer to the time of the assay have a greater influence on the HbA1c level.
The approximate relationship between HbA1c and mean blood glucose values during the preceding 2 to 3 months was analyzed in several studies. A recent study obtained the following correlation:
IFCC standardization (recalculated acc. to ref. 8)
• Estimated average glucose [mmol/L] = 0.146 x HbA1c (mmol/mol) + 0.834 or
• Estimated average glucose [mg/dL] = 2.64 x HbA1c (mmol/mol) + 15.03
Standardization acc. to DCCT/NGSP
• Estimated average glucose [mmol/L] = 1.59 x HbA1c (%) - 2.59 or
• Estimated average glucose [mg/dL] = 28.7 x HbA1c (%) - 46.7
The risk of diabetic complications, such as diabetic nephropathy and retinopathy, increases with poor metabolic control. In accordance with its function as an indicator for the mean blood glucose level, HbA1c predicts the development of diabetic complications in diabetes patients.
For monitoring long term glycemic control, testing every 3 to 4 months is generally sufficient. In certain clinical situations, such as gestational diabetes, or after a major change in therapy, it may be useful to measure HbA1c in 2 to 4 week intervals.
R1 | Antibody Reagent MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c antibody (ovine serum): ≥ 0.5 mg/mL; detergent; stabilizers; preservatives |
R3 | Polyhapten Reagent MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c polyhapten: ≥ 8 µg/mL; detergent; stabilizers; preservatives |
R1 is in position A and R3 is in position C. Position B contains H2O for technical reasons.
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.
This kit contains components classified as follows in accordance with the Regulation (EC) No. 1272/2008:
|
Warning
H317 | May cause an allergic skin reaction. |
Prevention:
P261 | Avoid breathing dust/fume/gas/mist/vapours/spray. |
P272 | Contaminated work clothing should not be allowed out of the workplace. |
P280 | Wear protective gloves. |
Response:
P333 + P313 | If skin irritation or rash occurs: Get medical advice/attention. |
P362 + P364 | Take off contaminated clothing and wash it before reuse. |
Disposal:
P501 | Dispose of contents/container to an approved waste disposal plant. |
Product safety labeling follows EU GHS guidance.
Contact phone: all countries: +49-621-7590
Quality control for Whole Blood and Hemolysate Application
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.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Anticoagulated venous or capillary blood or hemolysate.
The only acceptable anticoagulants are Li‑heparin, K2‑EDTA, K3‑EDTA, Fluoride/Na2‑EDTA, Na‑Heparin and Fluoride/potassium oxalate.
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.
See the limitations and interferences section for details about possible sample interferences.
Stability: | 3 days at 15‑25 °C 7 days at 2‑8 °C 6 months at (-15)‑(-25) °C |
Freeze only once. Mix specimen thoroughly after thawing.
1. Allow blood specimen and Hemolyzing Reagent for Tina‑quant HbA1c to equilibrate at room temperature before use.
2. Moderately mix the sample immediately prior to pipetting to ensure a homogeneous mixture of erythrocytes. Take care to avoid the formation of foam.
3. Dilute the sample with Hemolyzing Reagent for Tina‑quant HbA1c (Cat. No. 11488457 122) in the ratio 1:101 (1+100) using one of the following pipetting schemes.
Pipette into tubes:
HbA1c Hemolyzing Reagent for Tina‑quant HbA1c | 500 µL | 1000 µL | 2000 µL |
Specimen (patient or control) | 5 µL | 10 µL | 20 µL |
4. Mix using a vibration mixer or by gentle swirling.
5. The hemolysate can be used after the solution has changed color from red to brownish-green (approx. 1‑2 min).
Stability of the hemolysate: | 4 hours at 15‑25 °C 24 hours at 2‑8 °C 6 months at (-15)‑(-25) °C |
In vitro test for the quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in whole blood or in hemolysate on Roche/Hitachi cobas c systems.
This method uses TTABa) as the detergent in the hemolyzing reagent to eliminate interference from leukocytes (TTAB does not lyse leukocytes). Sample pretreatment to remove labile HbA1c is not necessary.
All hemoglobin variants which are glycated at the β‑chain N‑terminus and which have antibody-recognizable regions identical to that of HbA1c are determined by this assay. Consequently, the metabolic state of patients having uremia or the most frequent hemoglobinopathies (HbAS, HbAC, HbAE) can be determined using this assay.
a) TTAB = Tetradecyltrimethylammonium bromide
Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
Sample and addition of R1 (buffer/antibody)
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, formation of insoluble complexes does not take place.
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibody-polyhapten complex which can be determined turbidimetrically.
Hemoglobin
Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently not necessary.
The final result is expressed as mmol/mol HbA1c or % HbA1c and is calculated from the HbA1c/Hb ratio as follows:
Protocol 1 (mmol/mol HbA1c acc. to IFCC):
HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
Protocol 2 (% HbA1c acc. to DCCT/NGSP):
HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
Measuring range
Hemoglobin: 2.48‑24.8 mmol/L (4‑40 g/dL).
HbA1c: 0.186‑1.61 mmol/L (0.3‑2.6 g/dL)
It is recommended to switch the auto rerun function off.
Lower limits of measurement
Limit of Blank and Limit of Detection
Hemoglobin:
Limit of Blank = 0.31 mmol/L (0.50 g/dL)
Limit of Detection = 0.62 mmol/L (1.00 g/dL)
HbA1c:
Limit of Blank = 0.12 mmol/L (0.19 g/dL)
Limit of Detection = 0.18 mmol/L (0.29 g/dL)
The Limit of Blank and Limit of Detection were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A 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 %).
Protocol 1 (mmol/mol HbA1c acc. to IFCC):
20‑42 mmol/mol HbA1c
Protocol 2 (% HbA1c acc. to DCCT/NGSP): 4.0‑6.0 % HbA1c
HbA1c levels above the established reference range are an indication of hyperglycemia during the preceding 2 to 3 months or longer.
HbA1c levels may reach 195 mmol/mol (IFCC) or 20 % (DCCT/NGSP) or higher in poorly controlled diabetes. Therapeutic action is suggested at levels above 64 mmol/mol HbA1c (IFCC) or 8 % HbA1c (DCCT/NGSP). Diabetes patients with HbA1c levels below 53 mmol/mol HbA1c (IFCC) or 7 % HbA1c (DCCT/NGSP) meet the goal of the American Diabetes Association.
HbA1c levels below the established reference range may indicate recent episodes of hypoglycemia, the presence of Hb variants, or shortened lifetime of erythrocytes.
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
For diagnostic purposes, mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) should be used in conjunction with information from other diagnostic procedures and clinical evaluations.
The test is designed only for accurate and precise measurement of mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP). The individual results for total Hb and HbA1c concentration should not be reported.
The test is not intended for the diagnosis of diabetes mellitus or for judging day-to-day glucose control and should not be used to replace daily home testing of urine or blood glucose.
As a matter of principle, care must be taken when interpreting any HbA1c result from patients with Hb variants. Abnormal hemoglobins might affect the half life of the red cells or the in vivo glycation rates. In these cases even analytically correct results do not reflect the same level of glycemic control that would be expected in patients with normal hemoglobin.
Any cause of shortened erythrocyte survival will reduce exposure of erythrocytes to glucose with a consequent decrease in mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP), even though the time-averaged blood glucose level may be elevated. Causes of shortened erythrocyte lifetime might be hemolytic anemia or other hemolytic diseases, homozygous sickle cell trait, pregnancy, recent significant or chronic blood loss, etc. Caution should be used when interpreting the HbA1c results from patients with these conditions.
Glycated HbF is not detected as it does not contain the glycated β‑chain that characterizes HbA1c. However, HbF is measured in the Total Hb assay and as a consequence, specimens containing high amounts of HbF (> 10 %) may result in lower than expected mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).
Criterion: Recovery within ± 10 % of initial value.
Icterus:
Lipemia (Intralipid):
Drugs: No interference was found at therapeutic concentrations using common drug panels.
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated hemoglobin, glycated albumin and labile HbA1c were found for the anti-HbA1c antibodies used in this kit.
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.
| Analyzer(s) on which cobas c pack(s) can be used | ||
05336163 190 | Tina-quant Hemoglobin A1c Gen.3 (150 tests) | System ID 07 7455 3 | Roche/Hitachi cobas c 311, cobas c 501/502 |
04528417 190 | C.f.a.s. HbA1c (3 x 2 mL) | Code 674 | |
05479207 190 | PreciControl HbA1c norm (4 × 1 mL) | Code 208 | |
05912504 190 | PreciControl HbA1c path (4 × 1 mL) | Code 209 | |
04528182 190 | Hemolyzing Reagent Gen.2 (51 mL)* | System ID 07 6873 1 | |
11488457 122 | HbA1c Hemolyzing Reagent for Tina‑quant HbA1c (1000 mL) | For Hemolysate Application only |
* The value encoded in the instrument settings is 45 mL to account for the dead volume of the bottles.
Whole Blood Application - Standardized according to IFCC | ||
HB-W3: | ACN 871 | Hemoglobin (Hb) |
A1-W3: | ACN 881 | Hemoglobin A1c (HbA1c) |
RWD3: | ACN 891 | Ratio % HbA1c (acc. to DCCT/NGSP) |
A1CD2: | ACN 952 | Hemolyzing reagent |
Hemolysate Application - Standardized according to IFCC transferable to DCCT/NGSP | ||
HB-H3: | ACN 841 | Hemoglobin (Hb) |
A1–H3: | ACN 851 | Hemoglobin A1c (HbA1c) |
RHD3: | ACN 861 | Ratio % HbA1c (acc. to DCCT/NGSP) |
A1CD2: | ACN 952 | Hemolyzing reagent |
Ready for use
Whole Blood application for Hb (HB‑W3) and HbA1c (A1‑W3)
cobas c 311 test definition Hb (HB-W3) | |||
Assay type | 1-Point | ||
Reaction time / Assay points | 10 / 23 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
cobas c 311 test definition HbA1c (A1-W3) | |||
Assay type | 2-Point End | ||
Reaction time / Assay points | 10 / 23‑57 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
cobas c 501/502 test definition Hb (HB-W3) | |||
Assay type | 1-Point | ||
Reaction time / Assay points | 10 / 34 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
cobas c 501/502 test definition HbA1c (A1-W3) | |||
Assay type | 2-Point End | ||
Reaction time / Assay points | 10 / 34-70 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent (Hemolyzing reagent) | ||
Normal | 5 μL | 2 μL | 180 μL |
Decreased | 5 μL | 2 μL | 180 μL |
Increased | 5 μL | 2 μL | 180 μL |
Ratio definition for mmol/mol HbA1c and % HbA1c calculation | |
Abbreviated ratio name | RWI3 |
Equation | (A1-W3/HB-W3) × 1000 |
Unit | mmol/mol |
LREFHoelzel W, Weykamp C, Jeppsson JO, et al. IFCC Reference System for measurement of hemoglobin A1c in human blood and the National Standardization Schemes in the United States, Japan, and Sweden: a method comparison study. Clin Chem 2004;50:166-174. | |
Abbreviated ratio name | RWD3 (891) |
Equation | (A1-W3/HB-W3) × 91.5 + 2.15 |
Unit | % |
Protocol 2 is already implemented in the application (ACN 891). The mmol/mol HbA1c values according to Protocol 1 (IFCC) must be manually calculated according to the above equation. If requested a calculated test with the formula in Protocol 1 can be programmed under Utility > calculated test on the Roche/Hitachi cobas c 311 analyzer and on the Roche/Hitachi cobas c 501/502 analyzers. Please use the following settings:
Sample Type | Supernt. |
Unit of Measure | mM/M |
Report Name | HbA1c Gen.3 IFCC |
Item | RWI3 |
Formula | (A1-W3/HB-W3) x 1000 |
The ratio for HbA1c (mmol/mol HbA1c acc. to IFCC and % HbA1c acc. to DCCT/NGSP) will be automatically calculated after result output of both tests. It is recommended to report % HbA1c values (DCCT/NGSP) to one decimal place and mmol/mol HbA1c values (IFCC) without decimal places, which can be entered in the editable field “expected values”.
Hemolysate Application for Hb (HB‑H3) and HbA1c (A1‑H3)
cobas c 311 test definition Hb (HB-H3) | |||
Assay type | 1-Point | ||
Reaction time / Assay points | 10 / 23 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
cobas c 311 test definition HbA1c (A1-H3) | |||
Assay type | 2-Point End | ||
Reaction time / Assay points | 10 / 23-57 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
cobas c 501/502 test definition Hb (HB-H3) | |||
Assay type | 1‑Point | ||
Reaction time / Assay points | 10 / 34 | ||
Wavelength (sub/main) | 660 / 376 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
cobas c 501/502 test definition HbA1c (A1-H3) | |||
Assay type | 2‑Point End | ||
Reaction time / Assay points | 10 / 34-70 | ||
Wavelength (sub/main) | 660 / 340 nm | ||
Reaction direction | Increase | ||
Unit | mmol/L (g/dL) | ||
Reagent pipetting | Diluent (H2O) | ||
R1 | 120 μL | - | |
R3 | 24 μL | - | |
Sample volumes | Sample | Sample dilution | |
Sample | Diluent | ||
Normal | 5 μL | - | - |
Decreased | 5 μL | - | - |
Increased | 5 μL | - | - |
Ratio definition for HbA1c (mmol/mol (IFCC) or % (DCCT/NGSP))
calculation
Protocol 1 (mmol/mol HbA1c acc. to IFCC): | |
Abbreviated ratio name | RHI3 |
Equation | (A1-H3/HB-H3) x 1000 |
Unit | mmol/mol |
LREFHoelzel W, Weykamp C, Jeppsson JO, et al. IFCC Reference System for measurement of hemoglobin A1c in human blood and the National Standardization Schemes in the United States, Japan, and Sweden: a method comparison study. Clin Chem 2004;50:166-174. | |
Abbreviated ratio name | RHD3 (861) |
Equation | (A1-H3/HB-H3) x 91.5 + 2.15 |
Unit | % |
Protocol 2 is already implemented in the application (ACN 861). The mmol/mol HbA1c values according to Protocol 1 (IFCC) must be manually calculated according to the above equation. If requested a calculated test with the formula in Protocol 1 can be programmed under Utility > calculated test on the Roche/Hitachi cobas c 311 analyzer and on the Roche/Hitachi cobas c 501/502 analyzers. Please use the following settings:
Sample Type | Supernt. |
Unit of Measure | mM/M |
Report Name | HbA1c Gen.3 IFCC |
Item | RHI3 |
Formula | (A1-H3/HB-H3) x 1000 |
The ratio for HbA1c (mmol/mol HbA1c acc. to IFCC and % HbA1c acc. to DCCT/NGSP) will be automatically calculated after result output of both tests. It is recommended to report % HbA1c values (DCCT/NGSP) to one decimal place and mmol/mol HbA1c values (IFCC) without decimal places, which can be entered in the editable field “expected values”.
A1C-3 | |
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 |
Hemolyzing reagent | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label. |
When storing at temperatures under 3 °C, the reagent may become cloudy. This has no effect on the function of the reagent and is reversible at higher temperatures. It is therefore recommended to equilibrate the reagent at room temperature for approximately 10 minutes and mix thoroughly before use.
On-board in use and refrigerated on the analyzer: | 4 weeks |
Calibration for Whole Blood and Hemolysate Application
Hb | |
Calibrators | S1-S2: C.f.a.s. HbA1c |
Calibration mode | Linear |
HbA1c | |
Calibrators | S1-S6: C.f.a.s. HbA1c |
Calibration mode | Spline |
Calibration frequency | Hb and HbA1c: full calibration is recommended - after 29 days during shelf life - after reagent lot change - as required following quality control procedures |
Always calibrate both assays (Hb and HbA1c) in parallel. Automatic calibration at QC failure should be deactivated. |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against the approved IFCC reference method for the measurement of HbA1c in human blood
Note for Whole Blood and Hemolysate Application
Enter the assigned lot-specific and application-specific value of the calibrator. Use the appropriate C.f.a.s. HbA1c calibrator only.
The cobas c Hemolyzing Reagent Gen.2 pack, 51 mL, Cat. No. 04528182190, needs to be available on the analyzer otherwise the calibration cannot be performed.
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements with repeatability (n = 21) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained (data based on DCCT/NGSP values):
Whole Blood Application:
Repeatability | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.3 | 0.07 | 1.3 |
PreciControl HbA1c path | 9.9 | 0.11 | 1.1 |
Human sample 1 | 4.4 | 0.07 | 1.6 |
Human sample 2 | 5.6 | 0.09 | 1.6 |
Human sample 3 | 8.0 | 0.08 | 1.0 |
Human sample 4 | 10.6 | 0.11 | 1.1 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.3 | 0.08 | 1.4 |
PreciControl HbA1c path | 9.9 | 0.15 | 1.5 |
Human sample 1 | 4.4 | 0.09 | 1.9 |
Human sample 2 | 5.6 | 0.11 | 2.0 |
Human sample 3 | 8.0 | 0.11 | 1.4 |
Human sample 4 | 10.6 | 0.16 | 1.5 |
Hemolysate Application:
Repeatability | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.1 | 0.07 | 1.3 |
PreciControl HbA1c path | 10.2 | 0.10 | 1.0 |
Human sample 1 | 4.3 | 0.06 | 1.4 |
Human sample 2 | 5.6 | 0.07 | 1.2 |
Human sample 3 | 8.2 | 0.08 | 1.0 |
Human sample 4 | 10.9 | 0.11 | 1.0 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.1 | 0.11 | 2.2 |
PreciControl HbA1c path | 10.2 | 0.21 | 2.0 |
Human sample 1 | 4.3 | 0.10 | 2.3 |
Human sample 2 | 5.6 | 0.09 | 1.6 |
Human sample 3 | 8.2 | 0.16 | 1.9 |
Human sample 4 | 10.9 | 0.22 | 2.0 |
Evaluation of method comparison data is according to NGSP certification criteria. The mean difference between the two methods and the 95 % confidence intervals of the differences in the range from 4‑10 % (DCCT/NGSP) are given. 95 % of the differences between the values obtained for individual samples with both methods fall within the range defined by the lower and upper 95 % confidence intervals of the differences.
Whole Blood Application:
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the whole blood application (y) were compared
Sample size (n) = 80 | |
Mean difference | 0.07 % HbA1c |
Lower 95 % confidence interval of differences | -0.27 % HbA1c |
Upper 95 % confidence interval of differences | 0.42 % HbA1c |
The sample concentrations were between 4.7 % and 9.8 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the whole blood application (y) were compared
Sample size (n) = 82 | |
Mean difference | 0.07 % HbA1c |
Lower 95 % confidence interval of differences | -0.50 % HbA1c |
Upper 95 % confidence interval of differences | 0.65 % HbA1c |
The sample concentrations were between 5.0 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the whole blood application (y) were compared
Sample size (n) = 80 | |
Mean difference | -0.09 % HbA1c |
Lower 95 % confidence interval of differences | -0.46 % HbA1c |
Upper 95 % confidence interval of differences | 0.28 % HbA1c |
The sample concentrations were between 4.7 % and 9.8 % (DCCT/NGSP values). |
Hemolysate Application:
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the hemolysate application (y) were compared
Sample size (n) = 111 | |
Mean difference | -0.19 % HbA1c |
Lower 95 % confidence interval of differences | -0.52 % HbA1c |
Upper 95 % confidence interval of differences | 0.14 % HbA1c |
The sample concentrations were between 4.6 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the hemolysate application (y) were compared
Sample size (n) = 84 | |
Mean difference | -0.06 % HbA1c |
Lower 95 % confidence interval of differences | -0.53 % HbA1c |
Upper 95 % confidence interval of differences | 0.41 % HbA1c |
The sample concentrations were between 5.0 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the hemolysate application (y) were compared
Sample size (n) = 111 | |
Mean difference | -0.35 % HbA1c |
Lower 95 % confidence interval of differences | -0.68 % HbA1c |
Upper 95 % confidence interval of differences | -0.02 % HbA1c |
The sample concentrations were between 4.7 % and 9.9 % (DCCT/NGSP values). |
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a Roche/Hitachi cobas c 501 analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the hemolysate application (y) were compared
Sample size (n) = 113 | |
Mean difference | -0.10 % HbA1c |
Lower 95 % confidence interval of differences | -0.49 % HbA1c |
Upper 95 % confidence interval of differences | 0.31 % HbA1c |
The sample concentrations were between 4.8 % and 9.7 % (DCCT/NGSP values). |
Hemoglobin (Hb) consists of four protein subunits, each containing a heme moiety, and is the red-pigmented protein located in the erythrocytes. Its main function is to transport oxygen and carbon dioxide in blood. Each Hb molecule is able to bind four oxygen molecules. Hb consists of a variety of subfractions and derivatives. Among this heterogeneous group of hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction formed by the attachment of various sugars to the Hb molecule. HbA1c is formed in two steps by the nonenzymatic reaction of glucose with the N‑terminal amino group of the β‑chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This is rearranged to form stable HbA1c in a second reaction step.
In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte's life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. Glucose levels closer to the time of the assay have a greater influence on the HbA1c level.
The risk of diabetic complications, such as diabetic nephropathy and retinopathy, increases with poor metabolic control. In accordance with its function as an indicator for the mean blood glucose level, HbA1c predicts the development of diabetic complications in diabetes patients.
For routine clinical use, testing every 3 to 4 months is generally sufficient. In certain clinical situations, such as gestational diabetes, or after a major change in therapy, it may be useful to measure HbA1c in 2 to 4 week intervals.
R1 | Antibody Reagent MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c antibody (ovine serum): ≥ 0.5 mg/mL; detergent; stabilizers; preservatives |
R3 | Polyhapten Reagent MES buffer: 0.025 mol/L; TRIS buffer: 0.015 mol/L, pH 6.2; HbA1c polyhapten: ≥ 8 µg/mL; detergent; stabilizers; preservatives |
Aqueous buffered matrix, pH 7.25; tetradecyltrimethylammonium bromide: 36 g/L; sodium dihydrogenphosphate monohydrate: 16 mmol/L; sodium monohydrogenphosphate dihydrate: 64 mmol/L; stabilizers; preservatives |
R1 is in position A and R3 is in position C. Position B contains H2O for technical reasons.
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.
Quality control for Whole Blood and Hemolysate Application
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.
Follow the applicable government regulations and local guidelines for quality control.
When using Na-heparin, NaF/potassium oxalate, or NaF/sodium EDTA with the whole blood application, fresh whole blood samples should be analyzed. Do not use whole blood stored > 8 hours. Hemolysate should be prepared and stored for analysis if whole blood cannot be analyzed within 8 hours of collection. |
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Anticoagulated venous or capillary blood or hemolysate. The only acceptable anticoagulants are Li-heparin, Na-heparin, K2‑EDTA, K3‑EDTA, potassium fluoride/Na2‑EDTA, NaF/sodium EDTA and NaF/potassium oxalate.
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.
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.
3 days at 15‑25 °C 7 days at 2‑8 °C 6 months at (-15)‑(-25) °C |
Frozen stability of HbA1c has not been determined for samples treated with anticoagulants Na-heparin, NaF/potassium oxalate or NaF/sodium EDTA.
Freeze only once. Mix specimen thoroughly after thawing.
1. Allow blood specimen and Hemolyzing Reagent for Tina‑quant HbA1c to equilibrate at room temperature before use.
2. Moderately mix the sample immediately prior to pipetting to ensure a homogeneous mixture of erythrocytes. Take care to avoid the formation of foam.
3. Dilute the sample with Hemolyzing Reagent for Tina‑quant HbA1c (Cat. No. 11488457122) in the ratio 1:101 (1+100) using one of the following pipetting schemes.
Pipette into tubes:
HbA1c Hemolyzing Reagent for Tina‑quant HbA1c | 500 µL | 1000 µL | 2000 µL |
Specimen (patient or control) | 5 µL | 10 µL | 20 µL |
4. Mix using a vibration mixer or by gentle swirling.
5. The hemolysate can be used after the solution has changed color from red to brownish-green (approx. 1‑2 min).
4 hours at 15‑25 °C 24 hours at 2‑8 °C 6 months at (-15)‑(-25) °C |
Frozen stability of HbA1c has not been determined for samples treated with anticoagulants Na-heparin, NaF/potassium oxalate or NaF/sodium EDTA.
In vitro test for the quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in whole blood on Roche clinical chemistry analyzers. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus. Moreover, this test is to be used as an aid in diagnosis of diabetes and identifying patients who may be at risk for developing diabetes.
Test principle
The anticoagulated whole blood specimen is hemolyzed automatically on the COBAS INTEGRA 400 plus analyzer with COBAS INTEGRA Hemolyzing Reagent Gen.2. This method uses TTAB
All hemoglobin variants which are glycated at the β‑chain N‑terminus and which have antibody-recognizable regions identical to that of HbA1c are determined by this assay. Consequently, the metabolic state of diabetic patients having uremia or the most frequent hemoglobinopathies (HbAS, HbAC,
Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
Sample and addition of R1 (buffer/antibody):
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, complex formation does not take place.
Addition of SR (buffer/polyhapten) and start of reaction:
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibody-polyhapten complex which can be determined turbidimetrically.
Hemoglobin
Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently not necessary.
The final result is expressed as mmol/mol HbA1c or % HbA1c and is calculated from the HbA1c/Hb ratio as follows:
Protocol 1 (mmol/mol HbA1c acc. to IFCC):
HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
Protocol 2 (% HbA1c acc. to DCCT/NGSP):
HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
Hb: 2.48‑24.8 mmol/L (4‑40 g/dL)
HbA1c: 0.186‑1.61 mmol/L (0.3‑2.6 g/dL)
This corresponds to a measuring range of 23‑196 mmol/mol HbA1c (IFCC) and 4.2‑20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L (13.2 g/dL).
Hb: | Limit of Blank | = 0.31 mmol/L (0.50 g/dL) |
Limit of Detection | = 0.62 mmol/L (1.00 g/dL) | |
HbA1c: | Limit of Blank | = 0.12 mmol/L (0.19 g/dL) |
Limit of Detection | = 0.18 mmol/L (0.29 g/dL) |
The Limit of Blank and Limit of Detection were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A 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 %).
Protocol 1 (mmol/mol HbA1c acc. to IFCC): 29‑42 mmol/mol HbA1c
Protocol 2 (% HbA1c acc. to DCCT/NGSP): 4.8‑5.9 % HbA1c
This reference range was obtained by measuring
HbA1c levels may reach 195 mmol/mol (IFCC) or 20 % (DCCT/NGSP) or higher in poorly controlled diabetes. Therapeutic action is suggested at levels above 64 mmol/mol HbA1c (IFCC) or 8 % HbA1c (DCCT/NGSP). Diabetes patients with HbA1c levels below 53 mmol/mol HbA1c (IFCC) or 7 % HbA1c (DCCT/NGSP) meet the goal of the American Diabetes Association.
HbA1c levels below the established reference range may indicate recent episodes of hypoglycemia, the presence of Hb variants, or shortened lifetime of erythrocytes.
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Limitations - interference
For diagnostic purposes, mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) should be used in conjunction with information from other diagnostic procedures and clinical evaluations.
The test is designed only for accurate and precise measurement of mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP). The individual results for total Hb and HbA1c concentration should not be reported.
As a matter of principle, care must be taken when interpreting any HbA1c result from patients with Hb variants. Abnormal hemoglobins might affect the half life of the red cells or the in vivo glycation rates. In these cases even analytically correct results do not reflect the same level of glycemic control that would be expected in patients with normal hemoglobin.
Any cause of shortened erythrocyte survival or decrease in mean erythrocyte age will reduce exposure of erythrocytes to glucose with a consequent decrease in mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP), even though the time-averaged blood glucose level may be elevated. Causes of shortened erythrocyte lifetime might be hemolytic anemia or other hemolytic diseases, homozygous sickle cell trait, pregnancy, recent significant or chronic blood loss, etc. Similarly, recent blood transfusions can alter the mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP). Caution should be used when interpreting the HbA1c results from patients with these conditions. HbA1c must not be used for the diagnosis of diabetes mellitus in the presence of such conditions.
Glycated HbF is not detected as it does not contain the glycated β‑chain that characterizes HbA1c. However, HbF is measured in the Total Hb assay and as a consequence, specimens containing high amounts of HbF (> 10 %) may result in lower than expected mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).
mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) are not suitable for diagnosis of gestational diabetes.
In very rare cases of rapidly evolving type 1 diabetes the increase of HbA1c values might be delayed compared to the acute increase in glucose concentrations. In these conditions diabetes mellitus must be diagnosed based on plasma glucose concentrations and/or the typical clinical symptoms.
Criterion: Recovery within ± 10 % of initial value.
Icterus: No significant interference up to a conjugated and unconjugated bilirubin concentration of 1026 μmol/L or 60 mg/dL.
Lipemia (Intralipid): No significant interference up to an Intralipid concentrationof 600 mg/dL. There is poor correlation between the triglycerides concentration and turbidity.
Glycemia: No significant interference up to a glucose concentration of 55.5 mmol/L or 1000 mg/dL. A fasting sample is not required.
Rheumatoid factors: No significant interference up to a rheumatoid factor concentration of 750 IU/mL.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated hemoglobin, carbamylated hemoglobin, glycated albumin and labile HbA1c were found for the anti-HbA1c antibodies used in this kit.
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.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
05336163 190 | Tina-quant Hemoglobin A1c Gen.3 (150 tests) | System-ID 07 7455 3 | COBAS INTEGRA 400 plus |
Materials required (but not provided):
04528417 190 | C.f.a.s. HbA1c (3 × 2 mL) | System-ID 07 6852 9 | |
05479207 190 | PreciControl HbA1c norm (4 × 1 mL) | System-ID 07 7477 4 | |
05912504 190 | PreciControl HbA1c path (4 × 1 mL) | System-ID 07 7478 2 | |
04528328 190 | COBAS INTEGRA Hemolyzing Reagent Gen.2 (6 × 10 mL) | System-ID 07 6851 0 |
Multitest A1CW3, test ID 0‑264
Test HB‑W3, test ID 0‑265; test A1‑W3, test ID 0‑266
Ratio RWD3, test ID 0‑275 (% HbA1c acc. to DCCT/NGSP)
Ratio RWI3, test ID 0‑274 (mmol/mol HbA1c acc. to IFCC)
Profile PA1W3, test ID 0‑273
Ready for use
Abbreviated test name | HB-W3 |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction mode | R1-S |
Reaction direction | Increase |
Wavelength A/B | 378/659 nm |
Calc. first/last | 17/33 |
Predilution factor | 100 |
Unit | mmol/L |
Hb | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
Total volume | 126 µL |
Abbreviated test name | A1-W3 |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction mode | R1-S-SR |
Reaction direction | Increase |
Wavelength A/B | 340/659 nm |
Calc. first/last | 33/57 |
Predilution factor | 100 |
Unit | mmol/L |
HbA1c | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
SR | 24 µL | 0 µL |
Total volume | 150 µL |
Protocol 1 (mmol/mol HbA1c acc. to IFCC): | |
Abbreviated ratio name | RWI3 (0-274) |
Equation | (A1-W3/HB-W3) × 1000 |
Unit | mM/M |
Protocol 2 (% HbA1c acc. to DCCT/NGSP): | |
Abbreviated ratio name | RWD3 (0-275) |
Equation | (A1-W3/HB-W3) × 91.5 + 2.15 |
Unit | % |
Use the predefined profile (PA1W3, 0‑273) for simultaneous order entry of Hb (HB‑W3) and HbA1c (A1‑W3) tests from the same sample.
The ratio for HbA1c (mmol/mol HbA1c acc. to IFCC and % HbA1c acc. to DCCT/NGSP) will be automatically calculated after result output of both tests.
For dual reporting of both mmol/mol HbA1c (IFCC) units as well as % HbA1c (DCCT/NGSP) units please ensure that both ratio tests 0‑274 (acc. to IFCC) and 0‑275 (acc. to DCCT/NGSP) are activated.
Shelf life at 2‑8 °C | See expiration date on cobas c pack label | |
COBAS INTEGRA 400 plus system | ||
On-board in use at 10‑15 °C | 4 weeks |
Reagent cannot be frozen.
If freezing of a cassette is suspected a control measurement with this cassette is recommended.
Shelf life at 2‑8 °C | See expiration date on bottle label | |
COBAS INTEGRA 400 plus system | ||
On-board in use, ISE rack, closed bottles | 4 weeks | |
On-board in use, multi rack, open bottles | 2 days |
When storing at temperatures under 3 °C, the reagent may become cloudy. This has no effect on the function of the reagent and is reversible at higher temperatures. It is therefore recommended to equilibrate the reagent at room temperature for approximately 10 minutes and mix thoroughly before use.
Calibrator | C.f.a.s. HbA1c |
Calibrator | C.f.a.s. HbA1c |
Calibration dilution ratio | 1:1, 1:1.5, 1:2.1, 1:3, 1:6, 1:15, performed automatically by the instrument |
Calibrator diluent | COBAS INTEGRA Hemolyzing Reagent Gen.2, Cat. No. 04528328190 |
Calibration mode | Spline |
Calibration replicate | Duplicate recommended |
Calibration interval | Each lot, every 29 days, 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 approved IFCC reference method for the measurement of HbA1c in human blood
Enter the assigned lot-specific and application-specific value of the calibrator. Use the appropriate C.f.a.s. HbA1c calibrator only. COBAS INTEGRA Hemolyzing Reagent Gen.2, 6 × 11 mL, Cat. No. 04528328190, system‑ID 07 6851 0, needs to be available on the analyzer. Otherwise the calibration cannot be carried out.
Representative performance data on the COBAS INTEGRA analyzers are given below. Results obtained in individual laboratories may differ.
Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained (data based on DCCT/NGSP values):
Repeatability | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.81 | 0.09 | 1.6 |
PreciControl HbA1c path | 10.6 | 0.09 | 0.8 |
Human sample 1 | 5.37 | 0.09 | 1.7 |
Human sample 2 | 6.43 | 0.08 | 1.3 |
Human sample 3 | 7.50 | 0.10 | 1.4 |
Human sample 4 | 8.52 | 0.08 | 0.9 |
Human sample 5 | 10.9 | 0.09 | 0.8 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.74 | 0.12 | 2.1 |
PreciControl HbA1c path | 10.6 | 0.18 | 1.7 |
Human sample 1 | 5.35 | 0.12 | 2.3 |
Human sample 2 | 6.33 | 0.12 | 1.9 |
Human sample 3 | 7.50 | 0.14 | 1.8 |
Human sample 4 | 8.52 | 0.12 | 1.4 |
Human sample 5 | 10.7 | 0.21 | 1.9 |
Evaluation of method comparison data is according to former NGSP certification criteria. The mean difference between the two methods and the 95 % confidence intervals of the differences in the range from 4‑10 % (DCCT/NGSP) are given. 95 % of the differences between the values obtained for individual samples with both methods fall within the range defined by the lower and upper 95 % confidence intervals of the differences.
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a COBAS INTEGRA 400 plus analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the whole blood application (y) were compared with those determined using the same reagent with the whole blood application on a cobas c 501 analyzer (x).
Sample size (n) = 96 |
Mean difference: 0.03 % HbA1c |
Lower 95 % confidence interval of differences: −0.45 % HbA1c |
Upper 95 % confidence interval of differences: 0.51 % HbA1c |
The sample concentrations were between 4.30 and 11.7 % HbA1c (DCCT/NGSP).
Summary
Hemoglobin (Hb) consists of four protein subunits, each containing a heme moiety, and is the red-pigmented protein located in the erythrocytes. Its main function is to transport oxygen and carbon dioxide in blood. Each Hb molecule is able to bind four oxygen molecules. Hb consists of a variety of subfractions and derivatives. Among this heterogeneous group of hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction formed by the attachment of various sugars to the Hb molecule. HbA1c is formed in two steps by the nonenzymatic reaction of glucose with the N‑terminal amino group of the β‑chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This is rearranged to form stable HbA1c in a second reaction step.
In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte’s life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. Glucose levels closer to the time of the assay have a greater influence on the HbA1c level.
The approximate relationship between HbA1c and mean blood glucose values during the preceding 2 to 3 months was analyzed in several studies. A recent study obtained the following correlation:
IFCC standardization (recalculated acc. to ref. 8)
• Estimated average glucose [mmol/L] = 0.146 × HbA1c (mmol/mol) + 0.834
or
• Estimated average glucose [mg/dL] = 2.64 × HbA1c (mmol/mol) + 15.03
Standardization acc. to DCCT/NGSP
• Estimated average glucose [mmol/L] = 1.59 × HbA1c (%) - 2.59
or
• Estimated average glucose [mg/dL] = 28.7 × HbA1c (%) - 46.7
The risk of diabetic complications, such as diabetic nephropathy and retinopathy, increases with poor metabolic control. In accordance with its function as an indicator for the mean blood glucose level, HbA1c predicts the development of diabetic complications in diabetes patients.
For monitoring of long term glycemic control, testing every 3 to 4 months is generally sufficient. In certain clinical situations, such as gestational diabetes, or after a major change in therapy, it may be useful to measure HbA1c in 2 to 4 week intervals.
R1 | Antibody reagent FREFMES = 2-morpholinoethane sulfonic acid buffer: 0.025 mol/L; TRISFREFTRIS = Tris(hydroxymethyl)-aminomethane buffer: 0.015 mol/L, pH 6.2; HbA1c antibody (ovine serum): ≥ 0.5 mg/mL; detergents; stabilizers; preservative |
SR | Polyhapten reagent |
R1 is in position A and SR is in position C.
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.
This kit contains components classified as follows in accordance with the Regulation (EC) No. 1272/2008:
|
Warning
H317 | May cause an allergic skin reaction. |
Prevention:
P261 | Avoid breathing dust/fume/gas/mist/vapours/spray. |
P272 | Contaminated work clothing should not be allowed out of the workplace. |
P280 | Wear protective gloves. |
Response:
P333 + P313 | If skin irritation or rash occurs: Get medical advice/attention. |
P362 + P364 | Take off contaminated clothing and wash it before reuse. |
Disposal:
P501 | Dispose of contents/container to an approved waste disposal plant. |
Product safety labeling follows EU GHS guidance.
Contact phone: all countries: +49-621-7590
Quality control | PreciControl HbA1c norm |
PreciControl HbA1c path | |
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.
Follow the applicable government regulations and local guidelines for quality control.
Pretreat controls in the same way as samples.
HbA1c controls carry a declaration for mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP) only. No declarations for Hb and HbA1c concentrations are provided. As a consequence, HbA1c controls are handled like samples and cannot be included in the COBAS INTEGRA systems Quality Control Program.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Anticoagulated venous or capillary blood
The only acceptable anticoagulants are Li-Heparin, K2‑EDTA, K3‑EDTA, Fluoride/Na2‑EDTA, Na‑Heparin and Fluoride/potassium oxalate.
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.
See the limitations and interferences section for details about possible sample interferences.
Stability: | 3 days at 15‑25 °C |
7 days at 2‑8 °C | |
6 months at (‑15)‑(‑25) °C |
The recovery of HbA1c ratio values from sedimented samples, especially in case of poorly controlled diabetic patients, may be slightly elevated. To minimize this effect samples may be gently mixed by inversion prior to analysis.
Freeze only once. Mix specimen thoroughly after thawing.
In vitro test for the quantitative determination of mmol/mol hemoglobin A1c (IFCC) and % hemoglobin A1c (DCCT/NGSP) in hemolysate prepared from whole blood on Roche clinical chemistry analyzers. HbA1c determinations are useful for monitoring of long-term blood glucose control in individuals with diabetes mellitus. Moreover, this test is to be used as an aid in diagnosis of diabetes and identifying patients who may be at risk for developing diabetes.
Test principle
This method uses TTAB
All hemoglobin variants which are glycated at the β‑chain N‑terminus and which have antibody-recognizable regions identical to that of HbA1c are determined by this assay. Consequently, the metabolic state of diabetic patients having uremia or the most frequent hemoglobinopathies (HbAS, HbAC,
Hemoglobin A1c
The HbA1c determination is based on the turbidimetric inhibition immunoassay (TINIA) for hemolyzed whole blood.
Sample and addition of R1 (buffer/antibody):
Glycohemoglobin (HbA1c) in the sample reacts with anti-HbA1c antibody to form soluble antigen-antibody complexes. Since the specific HbA1c antibody site is present only once on the HbA1c molecule, complex formation does not take place.
Addition of SR (buffer/polyhapten) and start of reaction:
The polyhaptens react with excess anti-HbA1c antibodies to form an insoluble antibody-polyhapten complex which can be determined turbidimetrically.
Hemoglobin
Liberated hemoglobin in the hemolyzed sample is converted to a derivative having a characteristic absorption spectrum which is measured bichromatically during the preincubation phase (sample + R1) of the above immunological reaction. A separate Hb reagent is consequently not necessary.
The final result is expressed as mmol/mol HbA1c or % HbA1c and is calculated from the HbA1c/Hb ratio as follows:
Protocol 1 (mmol/mol HbA1c acc. to IFCC):
HbA1c (mmol/mol) = (HbA1c/Hb) × 1000
Protocol 2 (% HbA1c acc. to DCCT/NGSP):
HbA1c (%) = (HbA1c/Hb) × 91.5 + 2.15
Hb: 2.48‑24.8 mmol/L (4‑40 g/dL)
HbA1c: 0.186‑1.61 mmol/L (0.3‑2.6 g/dL)
This corresponds to a measuring range of 23‑196 mmol/mol HbA1c (IFCC) and 4.2‑20.1 % HbA1c (DCCT/NGSP) at a typical hemoglobin concentration of 8.2 mmol/L (13.2 g/dL).
Hb: | Limit of Blank | = 0.31 mmol/L (0.50 g/dL) |
Limit of Detection | = 0.62 mmol/L (1.00 g/dL) | |
HbA1c: | Limit of Blank | = 0.12 mmol/L (0.19 g/dL) |
Limit of Detection | = 0.18 mmol/L (0.29 g/dL) |
The Limit of Blank and Limit of Detection were determined in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP17‑A 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 %).
Protocol 1 (mmol/mol HbA1c acc. to IFCC): 29‑42 mmol/mol HbA1c
Protocol 2 (% HbA1c acc. to DCCT/NGSP): 4.8‑5.9 % HbA1c
This reference range was obtained by measuring
HbA1c levels may reach 195 mmol/mol (IFCC) or 20 % (DCCT/NGSP) or higher in poorly controlled diabetes. Therapeutic action is suggested at levels above 64 mmol/mol HbA1c (IFCC) or 8 % HbA1c (DCCT/NGSP). Diabetes patients with HbA1c levels below 53 mmol/mol HbA1c (IFCC) or 7 % HbA1c (DCCT/NGSP) meet the goal of the American Diabetes Association.
HbA1c levels below the established reference range may indicate recent episodes of hypoglycemia, the presence of Hb variants, or shortened lifetime of erythrocytes.
Each laboratory should investigate the transferability of the expected values to its own patient population and if necessary determine its own reference ranges.
Limitations - interference
For diagnostic purposes, mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) should be used in conjunction with information from other diagnostic procedures and clinical evaluations.
The test is designed only for accurate and precise measurement of mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP). The individual results for total Hb and HbA1c concentration should not be reported.
As a matter of principle, care must be taken when interpreting any HbA1c result from patients with Hb variants. Abnormal hemoglobins might affect the half life of the red cells or the in vivo glycation rates. In these cases even analytically correct results do not reflect the same level of glycemic control that would be expected in patients with normal hemoglobin.
Any cause of shortened erythrocyte survival or decrease in mean erythrocyte age will reduce exposure of erythrocytes to glucose with a consequent decrease in mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP), even though the time-averaged blood glucose level may be elevated. Causes of shortened erythrocyte lifetime might be hemolytic anemia or other hemolytic diseases, homozygous sickle cell trait, pregnancy, recent significant or chronic blood loss, etc. Similarly, recent blood transfusions can alter the mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP). Caution should be used when interpreting the HbA1c results from patients with these conditions. HbA1c must not be used for the diagnosis of diabetes mellitus in the presence of such conditions.
Glycated HbF is not detected as it does not contain the glycated β‑chain that characterizes HbA1c. However, HbF is measured in the Total Hb assay and as a consequence, specimens containing high amounts of HbF (> 10 %) may result in lower than expected mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP).
mmol/mol HbA1c values (IFCC) and % HbA1c values (DCCT/NGSP) are not suitable for diagnosis of gestational diabetes.
In very rare cases of rapidly evolving type 1 diabetes the increase of HbA1c values might be delayed compared to the acute increase in glucose concentrations. In these conditions diabetes mellitus must be diagnosed based on plasma glucose concentrations and/or the typical clinical symptoms.
Criterion: Recovery within ± 10 % of initial value.
Icterus: No significant interference up to a conjugated and unconjugated bilirubin concentration of 1026 μmol/L or 60 mg/dL.
Lipemia (Intralipid): No significant interference up to an Intralipid concentrationof 600 mg/dL. There is poor correlation between the triglycerides concentration and turbidity.
Glycemia: No significant interference up to a glucose concentration of 55.5 mmol/L or 1000 mg/dL. A fasting sample is not required.
Rheumatoid factors: No significant interference up to a rheumatoid factor concentration of 750 IU/mL.
Drugs: No interference was found at therapeutic concentrations using common drug panels.
Other: No cross reactions with HbA0, HbA1a, HbA1b, acetylated hemoglobin, carbamylated hemoglobin, glycated albumin and labile HbA1c were found for the anti-HbA1c antibodies used in this kit.
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.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
05336163190 | Tina-quant Hemoglobin A1c Gen.3 (150 tests) | System-ID 07 7455 3 | COBAS INTEGRA 400 plus |
04528417190 | C.f.a.s. HbA1c (3 × 2 mL) | System-ID 07 6852 9 | |
05479207190 | PreciControl HbA1c norm (4 × 1 mL) | System-ID 07 7477 4 | |
05912504190 | PreciControl HbA1c path (4 × 1 mL) | System-ID 07 7478 2 | |
11488457122 | Hemolyzing Reagent for Tina-quant HbA1c (1000 mL) | ||
04528328190 | COBAS INTEGRA Hemolyzing Reagent Gen.2 (6 × 10 mL) | System-ID 07 6851 0 |
Multitest A1CH3, test ID 0‑588
Test HB‑H3, test ID 0‑589; test A1‑H3, test ID 0‑590
Ratio RHD3, test ID 0‑622 (% HbA1c acc. to DCCT/NGSP)
Ratio RHI3, test ID 0‑621 (mmol/mol HbA1c acc. to IFCC)
Profile PA1H3, test ID 0‑620
Ready for use
Abbreviated test name | HB-H3 |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction mode | R1-S |
Reaction direction | Increase |
Wavelength A/B | 378/659 nm |
Calc. first/last | 17/33 |
Unit | mmol/L |
Hb | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
Total volume | 126 µL |
Abbreviated test name | A1-H3 |
Measuring mode | Absorbance |
Abs. calculation mode | Endpoint |
Reaction mode | R1-S-SR |
Reaction direction | Increase |
Wavelength A/B | 340/659 nm |
Calc. first/last | 33/57 |
Unit | mmol/L |
HbA1c | Diluent (H2O) | |
R1 | 120 µL | |
Sample | 6 µL | 0 µL |
SR | 24 µL | 0 µL |
Total volume | 150 µL |
Abbreviated ratio name | RHI3 (0-621) |
Equation | (A1-H3/HB-H3) × 1000 |
Unit | mM/M |
Abbreviated ratio name | RHD3 (0-622) |
Equation | (A1-H3/HB-H3) × 91.5 + 2.15 |
Unit | % |
Use the predefined profile (PA1H3, 0‑620) for simultaneous order entry of Hb (HB‑H3) and HbA1c (A1‑H3) tests from the same sample.
The ratio for HbA1c (mmol/mol HbA1c acc. to IFCC and % HbA1c acc. to DCCT/NGSP) will be automatically calculated after result output of both tests.
For dual reporting of both mmol/mol HbA1c (IFCC) units as well as % HbA1c (DCCT/NGSP) units please ensure that both ratio tests 0‑621 (acc. to IFCC) and 0‑620 (acc. to DCCT/NGSP) are activated.
Shelf life at 2‑8 °C | See expiration date on cobas c pack label | |
COBAS INTEGRA 400 plus system | ||
On-board in use at 10‑15 °C | 4 weeks |
Reagent cannot be frozen.
If freezing of a cassette is suspected a control measurement with this cassette is recommended.
Shelf life at 2‑8 °C | See expiration date on bottle label | |
COBAS INTEGRA 400 plus system | ||
On-board in use, ISE rack, closed bottles | 4 weeks | |
On-board in use, multi rack, open bottles | 2 days |
When storing at temperatures under 3 °C, the reagent may become cloudy. This has no effect on the function of the reagent and is reversible at higher temperatures. It is therefore recommended to equilibrate the reagent at room temperature for approximately 10 minutes and mix thoroughly before use.
Calibrator | C.f.a.s. HbA1c |
Calibrator | C.f.a.s. HbA1c |
Calibration dilution ratio | 1:1, 1:1.5, 1:2.1, 1:3, 1:6, 1:15, performed automatically by the instrument |
Calibrator diluent | COBAS INTEGRA Hemolyzing Reagent Gen.2, Cat. No. 04528328190 |
Calibration mode | Spline |
Calibration replicate | Duplicate recommended |
Calibration interval | Each lot, every 29 days, 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 approved IFCC reference method for the measurement of HbA1c in human blood
Enter the assigned lot-specific and application-specific value of the calibrator. Use the appropriate C.f.a.s. HbA1c calibrator only. COBAS INTEGRA Hemolyzing Reagent Gen.2, 6 × 11 mL, Cat. No. 04528328190, system‑ID 07 6851 0, needs to be available on the analyzer. Otherwise the calibration cannot be carried out.
Representative performance data on the COBAS INTEGRA analyzers are given below. Results obtained in individual laboratories may differ.
Repeatability and intermediate precision were determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP5 requirements (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained (data based on DCCT/NGSP values):
Repeatability | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.88 | 0.10 | 1.8 |
PreciControl HbA1c path | 10.6 | 0.07 | 0.7 |
Human sample 1 | 5.08 | 0.13 | 2.3 |
Human sample 2 | 6.51 | 0.13 | 2.0 |
Human sample 3 | 10.6 | 0.07 | 0.7 |
Human sample 4 | 7.57 | 0.08 | 1.1 |
Human sample 5 | 8.07 | 0.08 | 1.0 |
Intermediate precision | Mean | SD | CV |
|---|---|---|---|
PreciControl HbA1c norm | 5.88 | 0.12 | 2.0 |
PreciControl HbA1c path | 10.6 | 0.11 | 1.0 |
Human sample 1 | 5.08 | 0.12 | 2.6 |
Human sample 2 | 6.51 | 0.14 | 2.1 |
Human sample 3 | 10.6 | 0.12 | 1.1 |
Human sample 4 | 7.57 | 0.11 | 1.4 |
Human sample 5 | 7.98 | 0.09 | 1.1 |
Evaluation of method comparison data is according to former NGSP certification criteria. The mean difference between the two methods and the 95 % confidence intervals of the differences in the range from 4‑10 % (DCCT/NGSP) are given. 95 % of the differences between the values obtained for individual samples with both methods fall within the range defined by the lower and upper 95 % confidence intervals of the differences.
% HbA1c (DCCT/NGSP) values for human blood samples obtained on a COBAS INTEGRA 400 plus analyzer using the Tina‑quant Hemoglobin A1c Gen.3 reagent with the hemolysate application (y) were compared with those determined using the same reagent with the hemolysate application on a cobas c 501 analyzer (x).
Sample size (n) = 59 |
Mean difference: 0.16 % HbA1c |
Lower 95 % confidence interval of differences: −0.14 % HbA1c |
Upper 95 % confidence interval of differences: 0.45 % HbA1c |
The sample concentrations were between 4.31 and 12.0 % HbA1c (DCCT/NGSP).
Summary
Hemoglobin (Hb) consists of four protein subunits, each containing a heme moiety, and is the red-pigmented protein located in the erythrocytes. Its main function is to transport oxygen and carbon dioxide in blood. Each Hb molecule is able to bind four oxygen molecules. Hb consists of a variety of subfractions and derivatives. Among this heterogeneous group of hemoglobins HbA1c is one of the glycated hemoglobins, a subfraction formed by the attachment of various sugars to the Hb molecule. HbA1c is formed in two steps by the nonenzymatic reaction of glucose with the N‑terminal amino group of the β‑chain of normal adult Hb (HbA). The first step is reversible and yields labile HbA1c. This is rearranged to form stable HbA1c in a second reaction step.
In the erythrocytes, the relative amount of HbA converted to stable HbA1c increases with the average concentration of glucose in the blood. The conversion to stable HbA1c is limited by the erythrocyte’s life span of approximately 100 to 120 days. As a result, HbA1c reflects the average blood glucose level during the preceding 2 to 3 months. HbA1c is thus suitable to monitor long-term blood glucose control in individuals with diabetes mellitus. Glucose levels closer to the time of the assay have a greater influence on the HbA1c level.
The approximate relationship between HbA1c and mean blood glucose values during the preceding 2 to 3 months was analyzed in several studies. A recent study obtained the following correlation:
IFCC standardization (recalculated acc. to ref. 8)
• Estimated average glucose [mmol/L] = 0.146 × HbA1c (mmol/mol) + 0.834
or
• Estimated average glucose [mg/dL] = 2.64 × HbA1c (mmol/mol) + 15.03
Standardization acc. to DCCT/NGSP
• Estimated average glucose [mmol/L] = 1.59 × HbA1c (%) - 2.59
or
• Estimated average glucose [mg/dL] = 28.7 × HbA1c (%) - 46.7
The risk of diabetic complications, such as diabetic nephropathy and retinopathy, increases with poor metabolic control. In accordance with its function as an indicator for the mean blood glucose level, HbA1c predicts the development of diabetic complications in diabetes patients.
For monitoring of long term glycemic control, testing every 3 to 4 months is generally sufficient. In certain clinical situations, such as gestational diabetes, or after a major change in therapy, it may be useful to measure HbA1c in 2 to 4 week intervals.
R1 | Antibody reagent FREFMES = 2-morpholinoethane sulfonic acid buffer: 0.025 mol/L; TRISFREFTRIS = Tris(hydroxymethyl)-aminomethane buffer: 0.015 mol/L, pH 6.2; HbA1c antibody (ovine serum): ≥ 0.5 mg/mL; detergents; stabilizers; preservative |
SR | Polyhapten reagent |
R1 is in position A and SR is in position C.
Pay attention to all precautions and warnings listed in Section 1 / Introduction of this Method Manual.
This kit contains components classified as follows in accordance with the Regulation (EC) No. 1272/2008:
2-methyl-2H-isothiazol-3-one hydrochloride
EUH 208 | May produce an allergic reaction. |
Product safety labeling follows EU GHS guidance.
Quality control | PreciControl HbA1c norm |
PreciControl HbA1c path | |
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.
Follow the applicable government regulations and local guidelines for quality control.
Pretreat controls in the same way as samples.
HbA1c controls carry a declaration for mmol/mol HbA1c (IFCC) and % HbA1c (DCCT/NGSP) only. No declarations for Hb and HbA1c concentrations are provided. As a consequence, HbA1c controls are handled like samples and cannot be included in the COBAS INTEGRA systems Quality Control Program.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Anticoagulated venous or capillary blood or hemolysate
The only acceptable anticoagulants are Li‑Heparin, K2‑EDTA, K3‑EDTA, Fluoride/Na2‑EDTA, Na‑Heparin and Fluoride/potassium oxalate.
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.
See the limitations and interferences section for details about possible sample interferences.
3 days at 15‑25 °C | |
7 days at 2‑8 °C | |
6 months at (‑15)‑(‑25) °C |
Freeze only once. Mix specimen thoroughly after thawing.
Allow blood specimen and Hemolyzing Reagent for Tina‑quant HbA1c to equilibrate at room temperature before use.
Moderately mix the sample immediately prior to pipetting, to ensure homogeneous mixture of erythrocytes. Take care to avoid the formation of foam.
Dilute the sample with Hemolyzing Reagent for Tina‑quant HbA1c (Cat. No. 11488457122) in the ratio 1:101 (1+100) using one of the following pipetting schemes.
Pipette into tubes:
Hemolyzing Reagent for Tina‑quant HbA1c: 500 µL
specimen (patient or control): 5 µL
or
Hemolyzing Reagent for Tina‑quant HbA1c: 1000 µL
specimen (patient or control): 10 µL
or
Hemolyzing Reagent for Tina‑quant HbA1c: 2000 µL
specimen (patient or control): 20 µL
Mix using a vibration mixer or by gentle swirling.
The hemolysate can be used after the solution has changed color from red to brownish-green (approximately 1‑2 min).
4 hours at 15‑25 °C | |
24 hours at 2‑8 °C | |
6 months at (‑15)‑(‑25) °C |
Tina-quant Hemoglobin A1c Gen.3
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