ONLINE DAT Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative detection of benzodiazepines in human serum and plasma on Roche/Hitachi cobas c systems at a cutoff concentration of 200 ng/mL.
Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC‑MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC‑MS/MS) is the preferred confirmatory method.
The assay is based on the kinetic interaction of microparticles in a solution (KIMS)
When a serum sample contains the drug in question, this drug competes with the particle‑bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.
The presence of β‑glucuronidase enzyme enhances the Benzodiazepines II assay cross‑reactivity to some of the glucuronidated metabolites.
Qualitative assay
Results of this assay distinguish preliminary positive (≥ 200 ng/mL) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.
Criterion: No cross‑over at initial values of samples of 100 ng/mL and 300 ng/mL (control levels).
See the \"Specific performance data\" section of this document for information on substances tested with this assay. There is the possibility that other substances and/or factors may interfere with the test and cause erroneous results (e.g., technical or procedural errors).
A preliminary positive result with this assay indicates the presence of benzodiazepines and/or their metabolites in serum. It does not reflect the degree of intoxication.
Icterus:
Hemolysis:
Lipemia (Intralipid):
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 1200 IU/mL.
Immunoglobulins: No significant interference from immunoglobulins up to a concentration of 16 g/L (simulated by human immunoglobulin A), up to a concentration of 70 g/L (simulated by human immunoglobulin G) and up to a concentration of 10 g/L (simulated by human immunoglobulin M).
Albumin: No significant interference from human serum albumin up to a concentration of 70 g/L.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
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 | |
04939417190 | ONLINE DAT Benzodiazepines II (200 tests) | System‑ID 07 6997 5 | cobas c 501/502 |
Materials required (but not provided):
03304671190 | Preciset DAT Plus I calibrator CAL 5 | Code 435 | |
07978766190 | Serum DAT Control Low (ACQ Partner Channel*) | ||
07978740190 | Serum DAT Control High (ACQ Partner Channel*) | ||
04489357190 | NaCl Diluent 9 % (50 mL) | System‑ID 07 6869 3 |
*Roche does not hold the product registration for Partner Channels. The legal manufacturer indicated on the kit is solely responsible for all of the design, legal, and regulatory aspects of the product.
For cobas c 501 analyzer:
BEQ2S: ACN 603 (Serum/plasma): for qualitative assay, 200 ng/mL
For cobas c 502 analyzer:
BEQ2S: ACN 8603 (Serum/plasma): for qualitative assay, 200 ng/mL
Ready for use
Carefully invert reagent container several times prior to use to ensure that the reagent components are mixed.
Deselect Automatic Rerun for these applications in the Utility menu, Application screen, Range tab.
cobas c 501/502 test definition | |
Qualitative | |
Assay type | 2‑Point End |
Reaction time / Assay points | 10 / 16‑46 |
Wavelength (sub/main) | – /546 nm |
Reaction direction | Increase |
Unit | mAbs |
Reagent pipetting | |
R1 | 90 µL |
R2 | 40 µL |
Sample volumes | Sample |
200 ng/mL cutoff | |
Normal | 4.5 µL |
Decreased | 4.5 µL |
Increased | 4.5 µL |
BNZ2 | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 12 weeks |
Do not freeze. |
NaCl Diluent 9 % | |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 12 weeks |
Calibrators | Qualitative application |
200 ng/mL cutoff assay | |
S1: Preciset DAT Plus I calibrator ‑ CAL 5, 1000 ng/mL with automatic pre‑dilution | |
The drug concentration of the calibrator has been verified by GC‑MS. | |
Calibration K Factor | Enter the K Factor as -1000 into the Calibration menu, Status screen, Calibration Result window. |
Calibration mode | Qualitative application |
Linear | |
Calibration frequency | Blank calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against a primary reference method (GC‑MS).
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
A nordiazepam solution was added to 9 samples obtained from a human serum sample pool to achieve concentrations at approximately ‑100 %, ‑75 %, ‑50 %, ‑25 %, ±0 %, +25 %, +50 %, +75 %, and +100 % of the cutoff value. These samples were tested for precision. Following a CLSI (EP5‑A3) precision protocol, samples were tested in 2 replicates per run, 2 runs per day for 21 days, total n = 84. The following results were obtained on a
Drug | Concentration of Sample | Number of Determinations | Results |
Nordiazepam | zero drug | 84 | 84 Neg / 0 Pos |
Nordiazepam | ‑75 % | 84 | 84 Neg / 0 Pos |
Nordiazepam | ‑50 % | 84 | 84 Neg / 0 Pos |
Nordiazepam | ‑25 % | 84 | 0 Neg / 84 Pos |
Nordiazepam | cutoff | 84 | 0 Neg / 84 Pos |
Nordiazepam | +25 % | 84 | 0 Neg / 84 Pos |
Nordiazepam | +50 % | 84 | 0 Neg / 84 Pos |
Nordiazepam | +75 % | 83 | 0 Neg / 83 Pos |
Nordiazepam | +100 % | 84 | 0 Neg / 84 Pos |
The benzodiazepines constitute a class of versatile and widely prescribed central nervous system (CNS) depressant drugs with medically useful anxiolytic, sedative, hypnotic, muscle relaxant, and anticonvulsant activities.
Following ingestion, the benzodiazepines of the 1,4‑substituted class (including the triazolobenzodiazepine derivatives) are absorbed, metabolized, and excreted in the urine at different rates as a variety of structurally related metabolites. Metabolite diversity reflects the different physiochemical properties and metabolic pathways of the individual drugs. Overall metabolic similarities include removal of substituents from the β ring of the 1,4‑substituted benzodiazepines, α‑hydroxylation of the triazolobenzodiazepines, demethylation, hydroxylation of the three‑position carbon of the β ring, and conjugation of hydroxylated metabolites followed by urinary excretion predominantly as glucuronides.
R1 | Benzodiazepines antibody (sheep polyclonal); buffer; β‑glucuronidase enzyme; bovine serum albumin (BSA); 0.09 % sodium azide |
R2 | Conjugated benzodiazepine derivative microparticles; buffer; 0.09 % sodium azide |
R1 is in position A and R2 is in position B.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
Drug concentrations of the high and low controls have been verified by GC‑MS.
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.
Serum: Serum tubes with and without separating gel.
Plasma: K2‑ or K3‑EDTA, lithium heparin.
Stability: | 5 days capped at 15‑25 °C |
14 days capped at 2‑8 °C | |
6 months capped at -20 °C |
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Specimens can be repeatedly frozen and thawed up to 3 times.
Invert thawed specimens several times prior to testing.
CAUTION: Specimen dilutions should only be used to interpret results of Calc.? and Samp.? alarms, or when estimating concentration in preparation for GC‑MS or LC‑MS/MS. Dilution results are not intended for patient values. Dilution procedures, when used, should be validated.
Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on Roche/Hitachi cobas c systems at cutoff concentrations of 100 ng/mL, 200 ng/mL, and 300 ng/mL.
Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program.
Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC‑MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC‑MS/MS) is the preferred confirmatory method.
The assay is based on the kinetic interaction of microparticles in a solution (KIMS)
When a urine sample contains the drug in question, this drug competes with the particle‑bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.
The presence of β‑glucuronidase enzyme enhances the Benzodiazepines II assay cross‑reactivity to some of the glucuronidated metabolites.
Qualitative assay
Results of this assay distinguish preliminary positive (≥ 100 ng/mL, ≥ 200 ng/mL, or ≥ 300 ng/mL depending on the cutoff) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.
Semiquantitative assay
Results of this assay yield only approximate cumulative concentrations of the drug and its metabolites (see Analytical specificity section).
See the \"Specific performance data\" section of this document for information on substances tested with this assay. There is the possibility that other substances and/or factors may interfere with the test and cause erroneous results (e.g., technical or procedural errors).
A preliminary positive result with this assay indicates the presence of benzodiazepines and/or their metabolites in urine. It does not reflect the degree of intoxication.
Interfering substances were added to urine containing nordiazepam at ‑25 % and +25 % of the cutoff level at the concentration listed below. Samples were tested and the following results were obtained on a Roche/Hitachi 917 analyzer.
Semiquantitative (ng/mL) | 100 ng/mL | 200 ng/mL | 300 ng/mL | ||||
Compound | Cmpd. Conc. | Neg Level | Pos Level | Neg Level | Pos Level | Neg Level | Pos Level |
Acetone | 1 | 77(NEG) | 134(POS) | 157(NEG) | 260(POS) | 231(NEG) | 402(POS) |
Ascorbic acid | 1.5 | 78(NEG) | 132(POS) | 156(NEG) | 262(POS) | 233(NEG) | 399(POS) |
Conjugated bilirubin | 0.25 | 82(NEG) | 129(POS) | 156(NEG) | 247(POS) | 229(NEG) | 392(POS) |
Creatinine | 5 | 81(NEG) | 138(POS) | 158(NEG) | 259(POS) | 230(NEG) | 396(POS) |
Ethanol | 1 | 78(NEG) | 136(POS) | 151(NEG) | 261(POS) | 228(NEG) | 395(POS) |
Glucose | 20 | 81(NEG) | 138(POS) | 158(NEG) | 262(POS) | 236(NEG) | 403(POS) |
Hemoglobin | 1 | 76(NEG) | 139(POS) | 159(NEG) | 261(POS) | 228(NEG) | 398(POS) |
Human serum albumin | 5 | 83(NEG) | 140(POS) | 165(NEG) | 273(POS) | 243(NEG) | 422(POS) |
Oxalic acid | 2 | 74(NEG) | 128(POS) | 151(NEG) | 254(POS) | 226(NEG) | 388(POS) |
Sodium chloride | 0.5 | 79(NEG) | 139(POS) | 159(NEG) | 262(POS) | 234(NEG) | 389(POS) |
Urea | 6 | 80(NEG) | 138(POS) | 157(NEG) | 261(POS) | 233(NEG) | 405(POS) |
The same experiment was performed in the qualitative mode for each cutoff. All negative and positive samples recovered properly in the presence of the interfering substance.
An additional protocol was executed in which samples containing nordiazepam at control levels (±25 % of cutoff) with specific gravities ranging from 1.006 to 1.034 were tested. As with the other interferences, there were no control cross‑overs on any of the 3 assay cutoffs at either extreme specific gravity level.
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 | |
04939417190 | ONLINE DAT Benzodiazepines II (200 tests) | System‑ID 07 6997 5 |
Materials required (but not provided):
03304671190 | Preciset DAT Plus I calibrators CAL 1‑6 (6 x 5 mL) | Codes 431‑436 | |
03304680190 | Preciset DAT Plus II calibrators CAL 1‑6 (6 x 5 mL) | Codes 437‑442 | |
03304698190 | C.f.a.s. DAT Qualitative Plus (6 x 5 mL) | ||
04590856190 | C.f.a.s. DAT Qualitative Plus Clinical (3 x 5 mL) | Code 699 | |
03312950190 | Control Set DAT I (for 300 ng/mL assay) | ||
03312968190 | Control Set DAT II (for 100 ng/mL assay) | ||
04500873190 | Control Set DAT Clinical (for 100 ng/mL assay) | ||
03312976190 | Control Set DAT III (for 200 ng/mL assay) |
For cobas c 311/501 analyzers:
BZ2Q2: ACN 719 (Urine): for qualitative assay, 200 ng/mL
BZ3Q2: ACN 720 (Urine): for qualitative assay, 300 ng/mL
BZ1S2: ACN 728 (Urine): for semiquantitative assay, 100 ng/mL
BZ2S2: ACN 729 (Urine): for semiquantitative assay, 200 ng/mL
BZ3S2: ACN 730 (Urine): for semiquantitative assay, 300 ng/mL
BZQ1C: ACN 727 (Urine):
using C.f.a.s. DAT Qualitative Plus Clinical
For cobas c 502 analyzer:
BZ2Q2: ACN 8719 (Urine): for qualitative assay, 200 ng/mL
BZ3Q2: ACN 8720 (Urine): for qualitative assay, 300 ng/mL
BZ1S2: ACN 8728 (Urine): for semiquantitative assay, 100 ng/mL
BZ2S2: ACN 8729 (Urine): for semiquantitative assay, 200 ng/mL
BZ3S2: ACN 8730 (Urine): for semiquantitative assay, 300 ng/mL
BZQ1C: ACN 8727 (Urine):
using C.f.a.s. DAT Qualitative Plus Clinical
Ready for use
Carefully invert reagent container several times prior to use to ensure that the reagent components are mixed.
Deselect Automatic Rerun for these applications in the Utility menu, Application screen, Range tab.
cobas c 311 test definitions | |||
Semiquantitative | Qualitative | ||
Assay type | 2‑Point End | 2‑Point End | |
Reaction time / Assay points | 10 / 10‑31 | 10 / 10‑31 | |
Wavelength (sub/main) | – /546 nm | – /546 nm | |
Reaction direction | Increase | Increase | |
Unit | ng/mL | mAbs | |
Reagent pipetting | Diluent (H2O) | ||
R1 | 90 µL | – | |
R2 | 40 µL | – | |
Sample volumes | Sample | Sample dilution | |
100 and 200 ng/mL cutoffs | Sample | Diluent (NaCl) | |
Normal | 4.5 µL | – | – |
Decreased | 4.5 µL | – | – |
Increased | 4.5 µL | – | – |
300 ng/mL cutoff | |||
Normal | 2.0 µL | – | – |
Decreased | 2.0 µL | – | – |
Increased | 2.0 µL | – | – |
cobas c 501/502 test definitions | |||
Semiquantitative | Qualitative | ||
Assay type | 2‑Point End | 2‑Point End | |
Reaction time / Assay points | 10 / 16‑46 | 10 / 16‑46 | |
Wavelength (sub/main) | – /546 nm | – /546 nm | |
Reaction direction | Increase | Increase | |
Unit | ng/mL | mAbs | |
Reagent pipetting | Diluent (H2O) | ||
R1 | 90 µL | – | |
R2 | 40 µL | – | |
Sample volumes | Sample | Sample dilution | |
100 and 200 ng/mL cutoffs | Sample | Diluent (NaCl) | |
Normal | 4.5 µL | – | – |
Decreased | 4.5 µL | – | – |
Increased | 4.5 µL | – | – |
300 ng/mL cutoff | |||
Normal | 2.0 µL | – | – |
Decreased | 2.0 µL | – | – |
Increased | 2.0 µL | – | – |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 12 weeks |
Do not freeze. |
Calibrators | Semiquantitative applications |
100 and 200 ng/mL cutoff assays | |
S1‑6: Preciset DAT Plus II calibrators, CAL 1‑6 0, 50, 100, 200, 400, 1000 ng/mL | |
300 ng/mL cutoff assay | |
S1-6: Preciset DAT Plus I calibrators, CAL 1‑6 0, 150, 300, 600, 1000, 3000 ng/mL | |
Qualitative applications | |
100 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 3 (Test BZ1Q2) , 100 ng/mL, S1: C.f.a.s. DAT Qualitative Plus Clinical (Test BZQ1C) , 100 ng/mL | |
200 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 4, 200 ng/mL | |
300 ng/mL cutoff assay | |
S1: C.f.a.s. DAT Qualitative Plus or Preciset DAT Plus I calibrator - CAL 3, 300 ng/mL | |
The drug concentrations of the calibrators have been verified by GC‑MS. | |
Calibration K Factor | For the qualitative applications, enter the K Factor as -1000 into the Calibration menu, Status screen, Calibration Result window. |
Calibration mode | Semiquantitative applications |
Result Calculation Mode (RCM) FREFSee Results section. | |
Qualitative applications | |
Linear | |
Calibration frequency | Full (semiquantitative) or blank (qualitative) calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against a primary reference method (GC‑MS).
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
A nordiazepam solution was added to 5 samples obtained from a human urine sample pool to achieve low, high, and approximate positive and negative control concentrations of drug. These samples were tested for precision in qualitative and semiquantitative modes. Following a CLSI (EP5‑A2) precision protocol, samples were tested in 2 replicates per run, 2 runs per day for 10 days, total n = 40. The following results were obtained on a
Qualitative - 100 ng/mL Cutoff
Drug | Concentration of Sample, ng/mL | Number of Determinations | Results |
Nordiazepam | 57 | 40 | 40 Neg / 0 Pos |
Nordiazepam | 78 | 40 | 40 Neg / 0 Pos |
Nordiazepam | 124 | 40 | 0 Neg / 40 Pos |
Nordiazepam | 190 | 40 | 0 Neg / 40 Pos |
Nordiazepam | 414 | 40 | 0 Neg / 40 Pos |
Qualitative - 200 ng/mL Cutoff
Drug | Concentration of Sample, ng/mL | Number of Determinations | Results |
Nordiazepam | 102 | 40 | 40 Neg / 0 Pos |
Nordiazepam | 145 | 40 | 40 Neg / 0 Pos |
Nordiazepam | 241 | 40 | 0 Neg / 40 Pos |
Nordiazepam | 419 | 40 | 0 Neg / 40 Pos |
Nordiazepam | 842 | 40 | 0 Neg / 40 Pos |
Qualitative - 300 ng/mL Cutoff
Drug | Concentration of Sample, ng/mL | Number of Determinations | Results |
Nordiazepam | 148 | 40 | 40 Neg / 0 Pos |
Nordiazepam | 231 | 40 | 40 Neg / 0 Pos |
Nordiazepam | 364 | 40 | 0 Neg / 40 Pos |
Nordiazepam | 615 | 40 | 0 Neg / 40 Pos |
Nordiazepam | 1353 | 40 | 0 Neg / 40 Pos |
Semiquantitative - 100 ng/mL Cutoff
Drug | Sample Conc., ng/mL | Results# Neg / # Pos | Repeatability | Intermediate Precision | ||
SD, | CV, | SD, | CV, | |||
Nordiazepam | 57 | 40 / 0 | 1.9 | 3.4 | 2.0 | 3.5 |
Nordiazepam | 78 | 40 / 0 | 2.1 | 2.7 | 2.9 | 3.7 |
Nordiazepam | 124 | 0 / 40 | 1.6 | 1.3 | 3.8 | 3.1 |
Nordiazepam | 190 | 0 / 40 | 2.9 | 1.5 | 3.9 | 2.0 |
Nordiazepam | 414 | 0 / 40 | 3.6 | 0.9 | 9.9 | 2.4 |
Semiquantitative - 200 ng/mL Cutoff
Drug | Sample Conc., ng/mL | Results# Neg / # Pos | Repeatability | Intermediate Precision | ||
SD, | CV, | SD, | CV, | |||
Nordiazepam | 102 | 40 / 0 | 2.5 | 2.4 | 3.9 | 3.8 |
Nordiazepam | 145 | 40 / 0 | 1.8 | 1.3 | 5.4 | 3.7 |
Nordiazepam | 241 | 0 / 40 | 2.1 | 0.9 | 7.5 | 3.1 |
Nordiazepam | 419 | 0 / 40 | 6.6 | 1.6 | 12.2 | 2.9 |
Nordiazepam | 842 | 0 / 40 | 8.6 | 1.0 | 20.0 | 2.4 |
Semiquantitative - 300 ng/mL Cutoff
Drug | Sample Conc., ng/mL | Results# Neg / # Pos | Repeatability | Intermediate Precision | ||
SD, | CV, | SD, | CV, | |||
Nordiazepam | 148 | 40 / 0 | 3.7 | 2.5 | 4.8 | 3.3 |
Nordiazepam | 231 | 40 / 0 | 4.3 | 1.9 | 6.3 | 2.7 |
Nordiazepam | 364 | 0 / 40 | 4.5 | 1.2 | 8.2 | 2.3 |
Nordiazepam | 615 | 0 / 40 | 5.8 | 0.9 | 15.4 | 2.5 |
Nordiazepam | 1353 | 0 / 40 | 21.4 | 1.6 | 35.9 | 2.7 |
The benzodiazepines constitute a class of versatile and widely prescribed central nervous system (CNS) depressant drugs with medically useful anxiolytic, sedative, hypnotic, muscle relaxant, and anticonvulsant activities.
Following ingestion, the benzodiazepines of the 1,4‑substituted class (including the triazolobenzodiazepine derivatives) are absorbed, metabolized, and excreted in the urine at different rates as a variety of structurally related metabolites. Metabolite diversity reflects the different physiochemical properties and metabolic pathways of the individual drugs. Overall metabolic similarities include removal of substituents from the β ring of the 1,4‑substituted benzodiazepines, α‑hydroxylation of the triazolobenzodiazepines, demethylation, hydroxylation of the three‑position carbon of the β ring, and conjugation of hydroxylated metabolites followed by urinary excretion predominantly as glucuronides.
R1 | Benzodiazepines antibody (sheep polyclonal); buffer; β‑glucuronidase enzyme; bovine serum albumin (BSA); 0.09 % sodium azide |
R2 | Conjugated benzodiazepine derivative microparticles; buffer; 0.09 % sodium azide |
R1 is in position A and R2 is in position B.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
Drug concentrations of the controls have been verified by GC‑MS.
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.
Only the specimens listed below were tested and found acceptable.
Urine: Collect urine samples in clean glass or plastic containers. Fresh urine specimens do not require any special handling or pretreatment, but an effort should be made to keep pipetted samples free of gross debris. Samples should be within the normal physiological pH range of 5‑8. No additives or preservatives are required. It is recommended that urine specimens be stored at 2‑8 °C and tested within 5 days of collection.
For prolonged storage, freezing of samples is recommended.
Centrifuge highly turbid specimens before testing.
See the limitations and interferences section for details about possible sample interferences.
Adulteration or dilution of the sample can cause erroneous results. If adulteration is suspected, another sample should be collected. Specimen validity testing is required for specimens collected under the Mandatory Guidelines for Federal Workplace Drug Testing Programs.
CAUTION: Specimen dilutions should only be used to interpret results of Calc.? and Samp.? alarms, or when estimating concentration in preparation for GC‑MS or LC‑MS/MS. Dilution results are not intended for patient values. Dilution procedures, when used, should be validated.
Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on cobas c systems at cutoff concentrations of 100 ng/mL, 200 ng/mL, and 300 ng/mL.
Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program. Semiquantitative assays are intended to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas chromatography/mass spectrometry (GC-MS), or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS).
Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC‑MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC‑MS/MS) is the preferred confirmatory method.
The assay is based on the kinetic interaction of microparticles in a solution (KIMS)
When a urine sample contains the drug in question, this drug competes with the particle‑bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.
The presence of β‑glucuronidase enzyme enhances the Benzodiazepines II assay cross‑reactivity to some of the glucuronidated metabolites. Enzymatic cleavage makes the benzodiazepine part of the glucuronides more accessible for the antibody.
Qualitative assay
Results of this assay distinguish preliminary positive (≥ 100 ng/mL, ≥ 200 ng/mL, or ≥ 300 ng/mL depending on the cutoff) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.
Semiquantitative assay
Results of this assay yield only approximate cumulative concentrations of the drug and its metabolites (see \"Analytical specificity\" section).
See the \"Specific performance data\" section of this document for information on substances tested with this assay. There is the possibility that other substances and/or factors may interfere with the test and cause erroneous results (e.g., technical or procedural errors).
A preliminary positive result with this assay indicates the presence of benzodiazepines and/or their metabolites in urine. It does not reflect the degree of intoxication.
In very rare cases, gammopathy, in particular type IgM (Waldenström’s macroglobulinemia), may cause unreliable results.
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 | |
04939417190 | ONLINE DAT Benzodiazepines II (200 tests) | System‑ID 07 6997 5 | cobas c 311, cobas c 501/502 |
Materials required (but not provided):
03304671190 | Preciset DAT Plus I CAL 1‑6 (1 x 5 mL) | Codes 431‑436 | |
03304680190 | Preciset DAT Plus II CAL 1‑6 (1 x 5 mL) | Codes 437‑442 | |
03304698190 | C.f.a.s. DAT Qualitative Plus (6 x 5 mL) | ||
04590856190 | C.f.a.s. DAT Qualitative Plus Clinical (3 x 5 mL) | Code 699 | |
03312950190 | Control Set DAT I (for 300 ng/mL assay) | ||
03312968190 | Control Set DAT II (for 100 ng/mL assay) | ||
04500873190 | Control Set DAT Clinical (for 100 ng/mL assay) | ||
03312976190 | Control Set DAT III (for 200 ng/mL assay) |
For cobas c 311/501 analyzers:
BZ1Q2: ACN 718 (Urine): for qualitative assay, 100 ng/mL
BZ2Q2: ACN 719 (Urine): for qualitative assay, 200 ng/mL
BZ3Q2: ACN 720 (Urine): for qualitative assay, 300 ng/mL
BZ1S2: ACN 728 (Urine): for semiquantitative assay, 100 ng/mL
BZ2S2: ACN 729 (Urine): for semiquantitative assay, 200 ng/mL
BZ3S2: ACN 730 (Urine): for semiquantitative assay, 300 ng/mL
BZQ1C: ACN 727 (Urine): for qualitative assay, 100 ng/mL;
using C.f.a.s. DAT Qualitative Plus Clinical
For cobas c 502 analyzer:
BZ1Q2: ACN 8718 (Urine): for qualitative assay, 100 ng/mL
BZ2Q2: ACN 8719 (Urine): for qualitative assay, 200 ng/mL
BZ3Q2: ACN 8720 (Urine): for qualitative assay, 300 ng/mL
BZ1S2: ACN 8728 (Urine): for semiquantitative assay, 100 ng/mL
BZ2S2: ACN 8729 (Urine): for semiquantitative assay, 200 ng/mL
BZ3S2: ACN 8730 (Urine): for semiquantitative assay, 300 ng/mL
BZQ1C: ACN 8727 (Urine): for qualitative assay, 100 ng/mL;
using C.f.a.s. DAT Qualitative Plus Clinical
Ready for use
Carefully invert reagent container several times prior to use to ensure that the reagent components are mixed.
Deselect Automatic Rerun for these applications in the Utility menu, Application screen, Range tab.
cobas c 311 test definitions | |||
Semiquantitative | Qualitative | ||
Assay type | 2‑Point End | 2‑Point End | |
Reaction time / Assay points | 10 / 10‑31 | 10 / 10‑31 | |
Wavelength (sub/main) | – /546 nm | – /546 nm | |
Reaction direction | Increase | Increase | |
Unit | ng/mL | mAbs | |
Reagent pipetting | Diluent (H2O) | ||
R1 | 90 µL | – | |
R2 | 40 µL | – | |
Sample volumes | Sample | Sample dilution | |
100 and 200 ng/mL cutoffs | Sample | Diluent (NaCl) | |
Normal | 4.5 µL | – | – |
Decreased | 4.5 µL | – | – |
Increased | 4.5 µL | – | – |
300 ng/mL cutoff | |||
Normal | 2.0 µL | – | – |
Decreased | 2.0 µL | – | – |
Increased | 2.0 µL | – | – |
cobas c 501/502 test definitions | |||
Semiquantitative | Qualitative | ||
Assay type | 2‑Point End | 2‑Point End | |
Reaction time / Assay points | 10 / 16‑46 | 10 / 16‑46 | |
Wavelength (sub/main) | – /546 nm | – /546 nm | |
Reaction direction | Increase | Increase | |
Unit | ng/mL | mAbs | |
Reagent pipetting | Diluent (H2O) | ||
R1 | 90 µL | – | |
R2 | 40 µL | – | |
Sample volumes | Sample | Sample dilution | |
100 and 200 ng/mL cutoffs | Sample | Diluent (NaCl) | |
Normal | 4.5 µL | – | – |
Decreased | 4.5 µL | – | – |
Increased | 4.5 µL | – | – |
300 ng/mL cutoff | |||
Normal | 2.0 µL | – | – |
Decreased | 2.0 µL | – | – |
Increased | 2.0 µL | – | – |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 12 weeks |
Do not freeze. |
Calibrators | Semiquantitative applications |
100 and 200 ng/mL cutoff assays | |
S1‑6: Preciset DAT Plus II calibrators, CAL 1‑6 0, 50, 100, 200, 400, 1000 ng/mL | |
300 ng/mL cutoff assay | |
S1-6: Preciset DAT Plus I calibrators, CAL 1‑6 0, 150, 300, 600, 1000, 3000 ng/mL | |
Qualitative applications | |
100 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 3 | |
200 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 4, 200 ng/mL | |
300 ng/mL cutoff assay | |
S1: C.f.a.s. DAT Qualitative Plus or Preciset DAT Plus I calibrator - CAL 3, 300 ng/mL | |
The drug concentrations of the calibrators have been verified by GC‑MS. | |
Calibration K Factor | For the qualitative applications, enter the K Factor as -1000 into the Calibration menu, Status screen, Calibration Result window. |
Calibration mode | Semiquantitative applications |
Result Calculation Mode (RCM) FREFSee Results section. | |
Qualitative applications | |
Linear | |
Calibration frequency | Full (semiquantitative) or blank (qualitative) calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against a primary reference method (GC‑MS).
Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.
Results obtained in individual laboratories may differ due to heterogeneous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). Results for repeatability and intermediate precision were obtained on the cobas c 501 analyzer.
Semiquantitative precision - 100 ng/mL | |||
Repeatability | Mean | SD | CV |
Urine - 50 % | 45.0 | 2.56 | 5.7 |
Urine - 25 % | 68.8 | 2.44 | 3.6 |
DAT2N | 78.8 | 1.99 | 2.5 |
Cutoff urine | 99.7 | 2.38 | 2.4 |
Urine + 25 % | 123 | 2.43 | 2.0 |
DAT2P | 127 | 2.70 | 2.1 |
Urine + 50 % | 146 | 2.59 | 1.8 |
Intermediate precision | Mean | SD | CV |
Urine - 50 % | 45.0 | 2.79 | 6.2 |
Urine - 25 % | 68.8 | 2.65 | 3.9 |
DAT2N | 81.2 | 3.08 | 3.8 |
Cutoff urine | 99.7 | 2.63 | 2.6 |
Urine + 25 % | 123 | 2.92 | 2.4 |
DAT2P | 129 | 3.40 | 2.6 |
Urine + 50 % | 146 | 2.94 | 2.0 |
Cutoff (100) | Number | Correct | Confidence level |
Urine - 50 % | 84 | 84 | > 95 % negative reading |
Urine - 25 % | 84 | 84 | > 95 % negative reading |
DAT2N | 84 | 84 | > 95 % negative reading |
Cutoff urine | 84 | n.a.* | n.a.* |
Urine + 25 % | 84 | 84 | > 95 % positive reading |
DAT2P | 84 | 84 | > 95 % positive reading |
Urine + 50 % | 84 | 84 | > 95 % positive reading |
*n.a. = not applicable
Semiquantitative precision - 200 ng/mL | |||
Repeatability | Mean | SD | CV |
Urine - 50 % | 98.2 | 2.50 | 2.5 |
Urine - 25 % | 146 | 2.51 | 1.7 |
DAT3N | 152 | 2.70 | 1.8 |
Cutoff urine | 199 | 2.48 | 1.2 |
Urine + 25 % | 242 | 3.15 | 1.3 |
DAT3P | 250 | 2.67 | 1.1 |
Urine + 50 % | 279 | 2.34 | 0.8 |
Intermediate precision | Mean | SD | CV |
Urine - 50 % | 98.2 | 3.09 | 3.1 |
Urine - 25 % | 146 | 2.87 | 2.0 |
DAT3N | 150 | 3.49 | 2.3 |
Cutoff urine | 199 | 3.33 | 1.7 |
Urine + 25 % | 242 | 3.72 | 1.5 |
DAT3P | 248 | 5.68 | 2.3 |
Urine + 50 % | 279 | 4.88 | 1.7 |
Cutoff (200) | Number | Correct | Confidence level |
Urine - 50 % | 84 | 84 | > 95 % negative reading |
Urine - 25 % | 84 | 84 | > 95 % negative reading |
DAT3N | 84 | 84 | > 95 % negative reading |
Cutoff urine | 84 | n.a.* | n.a.* |
Urine + 25 % | 84 | 84 | > 95 % positive reading |
DAT3P | 84 | 84 | > 95 % positive reading |
Urine + 50 % | 84 | 84 | > 95 % positive reading |
*n.a. = not applicable
Semiquantitative precision - 300 ng/mL | |||
Repeatability | Mean | SD | CV |
Urine - 50 % | 151 | 4.41 | 2.9 |
Urine - 25 % | 211 | 3.99 | 1.9 |
DAT1N | 223 | 5.50 | 2.5 |
Cutoff urine | 276 | 4.17 | 1.5 |
DAT1P | 347 | 4.87 | 1.4 |
Urine + 25 % | 354 | 5.21 | 1.5 |
Urine + 50 % | 432 | 5.14 | 1.2 |
Intermediate precision | Mean | SD | CV |
Urine - 50 % | 151 | 5.31 | 3.5 |
Urine - 25 % | 211 | 5.40 | 2.6 |
DAT1N | 214 | 7.23 | 3.4 |
Cutoff urine | 276 | 6.07 | 2.2 |
DAT1P | 347 | 10.1 | 2.9 |
Urine + 25 % | 354 | 6.17 | 1.7 |
Urine + 50 % | 432 | 6.70 | 1.6 |
Cutoff (300) | Number | Correct | Confidence level |
Urine - 50 % | 84 | 84 | > 95 % negative reading |
Urine - 25 % | 84 | 84 | > 95 % negative reading |
DAT1N | 84 | 84 | > 95 % negative reading |
Cutoff urine | 84 | n.a.* | n.a.* |
DAT1P | 84 | 84 | > 95 % positive reading |
Urine + 25 % | 84 | 84 | > 95 % positive reading |
Urine + 50 % | 84 | 84 | > 95 % positive reading |
*n.a. = not applicable
The data obtained on cobas c 501 analyzer(s) are representative for cobas c 311 analyzer(s).
The benzodiazepines constitute a class of versatile and widely prescribed central nervous system (CNS) depressant drugs with medically useful anxiolytic, sedative, hypnotic, muscle relaxant, and anticonvulsant activities.
Following ingestion, the benzodiazepines of the 1,4‑substituted class (including the triazolobenzodiazepine derivatives) are absorbed, metabolized, and excreted in the urine at different rates as a variety of structurally related metabolites. Metabolite diversity reflects the different physiochemical properties and metabolic pathways of the individual drugs. Overall metabolic similarities include removal of substituents from the β ring of the 1,4‑substituted benzodiazepines, α‑hydroxylation of the triazolobenzodiazepines, demethylation, hydroxylation of the three‑position carbon of the β ring, and conjugation of hydroxylated metabolites followed by urinary excretion predominantly as glucuronides.
R1 | Benzodiazepines antibody (sheep polyclonal); buffer; β‑glucuronidase enzyme; bovine serum albumin (BSA); 0.09 % sodium azide |
R2 | Conjugated benzodiazepine derivative microparticles; buffer; 0.09 % sodium azide |
R1 is in position A and R2 is in position B.
For in vitro diagnostic use for health care professionals. Exercise the normal precautions required for handling all laboratory reagents.
Infectious or microbial waste:
Warning: handle waste as potentially biohazardous material. Dispose of waste according to accepted laboratory instructions and procedures.
Environmental hazards:
Apply all relevant local disposal regulations to determine the safe disposal.
Safety data sheet available for professional user on request.
For USA: Caution: Federal law restricts this device to sale by or on the order of a physician.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
Drug concentrations of Control Set DAT I, II, III, and Clinical have been verified by GC‑MS.
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.
Only the specimens listed below were tested and found acceptable.
Urine: Collect urine samples in clean glass or plastic containers. Fresh urine specimens do not require any special handling or pretreatment, but an effort should be made to keep pipetted samples free of gross debris. Samples should be within the normal physiological pH range of 5‑8. No additives or preservatives are required. It is recommended that urine specimens be stored at 2‑8 °C and tested within 5 days of collection.
Centrifuge highly turbid specimens before testing.
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.
Adulteration or dilution of the sample can cause erroneous results. If adulteration is suspected, another sample should be collected. Specimen validity testing is required for specimens collected under the Mandatory Guidelines for Federal Workplace Drug Testing Programs.
CAUTION: Specimen dilutions should only be used to interpret results of Calc.? and Samp.? alarms, or when estimating concentration in preparation for GC‑MS or LC‑MS/MS. Dilution results are not intended for patient values. Dilution procedures, when used, should be validated.
Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on Roche/Hitachi cobas c systems at cutoff concentrations of 100 ng/mL, 200 ng/mL, and 300 ng/mL.
Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program.
Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC‑MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC‑MS/MS) is the preferred confirmatory method.
The assay is based on the kinetic interaction of microparticles in a solution (KIMS)
When a urine sample contains the drug in question, this drug competes with the particle‑bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.
The presence of β‑glucuronidase enzyme enhances the Benzodiazepines II assay cross‑reactivity to some of the glucuronidated metabolites.
Qualitative assay
Results of this assay distinguish preliminary positive (≥ 100 ng/mL, ≥ 200 ng/mL, or ≥ 300 ng/mL depending on the cutoff) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.
Semiquantitative assay
Results of this assay yield only approximate cumulative concentrations of the drug and its metabolites (see Analytical specificity section).
See the \"Specific performance data\" section of this document for information on substances tested with this assay. There is the possibility that other substances and/or factors may interfere with the test and cause erroneous results (e.g., technical or procedural errors).
A preliminary positive result with this assay indicates the presence of benzodiazepines and/or their metabolites in urine. It does not reflect the degree of intoxication.
Interfering substances were added to urine containing nordiazepam at ‑25 % and +25 % of the cutoff level at the concentration listed below. Samples were tested and the following results were obtained on a Roche/Hitachi 917 analyzer.
Semiquantitative (ng/mL) | 100 ng/mL | 200 ng/mL | 300 ng/mL | ||||
Compound | Cmpd. conc. | Neg level | Pos level | Neg level | Pos level | Neg level | Pos level |
Acetone | 1 | 77(NEG) | 134(POS) | 157(NEG) | 260(POS) | 231(NEG) | 402(POS) |
Ascorbic acid | 1.5 | 78(NEG) | 132(POS) | 156(NEG) | 262(POS) | 233(NEG) | 399(POS) |
Conjugated bilirubin | 0.25 | 82(NEG) | 129(POS) | 156(NEG) | 247(POS) | 229(NEG) | 392(POS) |
Creatinine | 5 | 81(NEG) | 138(POS) | 158(NEG) | 259(POS) | 230(NEG) | 396(POS) |
Ethanol | 1 | 78(NEG) | 136(POS) | 151(NEG) | 261(POS) | 228(NEG) | 395(POS) |
Glucose | 20 | 81(NEG) | 138(POS) | 158(NEG) | 262(POS) | 236(NEG) | 403(POS) |
Hemoglobin | 1 | 76(NEG) | 139(POS) | 159(NEG) | 261(POS) | 228(NEG) | 398(POS) |
Human serum albumin | 5 | 83(NEG) | 140(POS) | 165(NEG) | 273(POS) | 243(NEG) | 422(POS) |
Oxalic acid | 2 | 74(NEG) | 128(POS) | 151(NEG) | 254(POS) | 226(NEG) | 388(POS) |
Sodium chloride | 0.5 | 79(NEG) | 139(POS) | 159(NEG) | 262(POS) | 234(NEG) | 389(POS) |
Urea | 6 | 80(NEG) | 138(POS) | 157(NEG) | 261(POS) | 233(NEG) | 405(POS) |
The same experiment was performed in the qualitative mode for each cutoff. All negative and positive samples recovered properly in the presence of the interfering substance.
An additional protocol was executed in which samples containing nordiazepam at control levels (±25 % of cutoff) with specific gravities ranging from 1.006 to 1.034 were tested. As with the other interferences, there were no control cross‑overs on any of the 3 assay cutoffs at either extreme specific gravity level.
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
|
| Analyzer(s) on which cobas c pack(s) can be used | |
08056927190 | ONLINE DAT Benzodiazepines II (850 tests) | System‑ID 2028 001 |
Materials required (but not provided):
03304671190 | Preciset DAT Plus I calibrators CAL 1‑6 (6 x 5 mL) | Codes 20431‑20436 | |
03304680190 | Preciset DAT Plus II calibrators CAL 1‑6 (6 x 5 mL) | Codes 20437‑20442 | |
03304698190 | C.f.a.s. DAT Qualitative Plus (6 x 5 mL) | Code 20698 | |
04590856190 | C.f.a.s. DAT Qualitative Plus Clinical (3 x 5 mL) | Code 20699 | |
03312950190 | Control Set DAT I (for 300 ng/mL assay) | ||
03312968190 | Control Set DAT II (for 100 ng/mL assay) | ||
04500873190 | Control Set DAT Clinical (for 100 ng/mL assay) | ||
03312976190 | Control Set DAT III (for 200 ng/mL assay) |
BZ1Q2: ACN 20280 (Urine): for qualitative assay, 100 ng/mL
BZ2Q2: ACN 20281 (Urine): for qualitative assay, 200 ng/mL
BZ3Q2: ACN 20282 (Urine): for qualitative assay, 300 ng/mL
BZ1S2: ACN 20284 (Urine): for semiquantitative assay, 100 ng/mL
BZ2S2: ACN 20285 (Urine): for semiquantitative assay, 200 ng/mL
BZ3S2: ACN 20286 (Urine): for semiquantitative assay, 300 ng/mL
BZQ1C: ACN 20283 (Urine): for qualitative assay, 100 ng/mL;
using C.f.a.s. DAT Qualitative Plus Clinical
BZ3‑QP: ACN 20288 (Urine): for qualitative assay, 300 ng/mL; using C.f.a.s. DAT Qualitative Plus
Ready for use
Carefully invert reagent container several times prior to use to ensure that the reagent components are mixed.
Semiquantitative | Qualitative | ||
Reporting time | 10 min | 10 min | |
Wavelength (sub/main) | – /546 nm | – /546 nm | |
Reagent pipetting | Diluent (H2O) | ||
R1 | 63 µL | – | |
R2 | 28 µL | – | |
Sample volumes | Sample | Sample dilution | |
100 and 200 ng/mL cutoffs | Sample | Diluent (NaCl) | |
Normal | 3.2 µL | – | – |
Decreased | 3.2 µL | – | – |
Increased | 3.2 µL | – | – |
300 ng/mL cutoff | |||
Normal | 1.4 µL | – | – |
Decreased | 1.4 µL | – | – |
Increased | 1.4 µL | – | – |
For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 26 weeks |
Do not freeze. |
Calibrators | Semiquantitative applications |
100 and 200 ng/mL cutoff assays | |
S1‑6: Preciset DAT Plus II calibrators, CAL 1‑6 0, 50, 100, 200, 400, 1000 ng/mL | |
300 ng/mL cutoff assay | |
S1-6: Preciset DAT Plus I calibrators, CAL 1‑6 0, 150, 300, 600, 1000, 3000 ng/mL | |
Qualitative applications | |
100 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 3 (Test BZ1Q2), 100 ng/mL, S1: C.f.a.s. DAT Qualitative Plus Clinical (Test BZQ1C), 100 ng/mL | |
200 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 4, 200 ng/mL | |
300 ng/mL cutoff assay | |
S1: Preciset DAT Plus I calibrator - CAL 3 (Test BZ3Q2), 300 ng/mL, S1: C.f.a.s. DAT Qualitative Plus (Test BZ3‑QP), 300 ng/mL | |
The drug concentrations of the calibrators have been verified by GC‑MS. | |
Calibration K factor | For the qualitative application a K factor of -1000 is predefined in the application settings. |
Calibration mode | Semiquantitative applications |
Non‑linear | |
Qualitative applications | |
Linear | |
Calibration frequency | Full calibration |
For the cutoff calibrator a value of \"0\" is encoded in the e‑barcode in order to ensure flagging of positive samples with >Test and negative absorbance values for negative samples.
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against a primary reference method (GC‑MS).
Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.
Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained:
Semiquantitative precision - 100 ng/mL cutoff | |||
Repeatability | Mean | SD | CV |
Urine -50 % | 49.5 | 3.30 | 6.7 |
DATCN | 79.5 | 3.10 | 3.9 |
DAT2N | 80.0 | 3.51 | 4.4 |
Cutoff urine | 105 | 3.68 | 3.5 |
DAT2P | 129 | 3.38 | 2.6 |
DATCP | 128 | 3.51 | 2.7 |
Urine +50 % | 153 | 2.98 | 1.9 |
Intermediate | Mean | SD | CV |
Urine -50 % | 49.5 | 4.00 | 8.1 |
DATCN | 79.5 | 4.09 | 5.1 |
DAT2N | 80.0 | 4.28 | 5.4 |
Cutoff urine | 105 | 4.82 | 4.6 |
DAT2P | 129 | 4.37 | 3.4 |
DATCP | 128 | 4.31 | 3.4 |
Urine +50 % | 153 | 4.58 | 3.0 |
Qualitative precision - 100 ng/mL cutoff | |||
Cutoff (100) | Number | Correct | Confidence level |
Urine -50 % | 84 | 84 | > 95 % negative reading |
DATCN | 84 | 84 | > 95 % negative reading |
DAT2N | 84 | 84 | > 95 % negative reading |
Cutoff urine | 84 | n.a.* | n.a.* |
DAT2P | 84 | 84 | > 95 % positive reading |
DATCP | 84 | 84 | > 95 % positive reading |
Urine +50 % | 84 | 84 | > 95 % positive reading |
*n.a. = not applicable
Semiquantitative precision - 200 ng/mL cutoff | |||
Repeatability | Mean | SD | CV |
Urine -50 % | 104 | 2.49 | 2.4 |
DAT3N | 152 | 3.78 | 2.5 |
Cutoff urine | 200 | 2.94 | 1.5 |
DAT3P | 249 | 3.61 | 1.4 |
Urine +50 % | 308 | 2.52 | 0.8 |
Intermediate | Mean | SD | CV |
Urine -50 % | 104 | 4.20 | 4.0 |
DAT3N | 152 | 4.31 | 2.8 |
Cutoff urine | 200 | 5.85 | 2.9 |
DAT3P | 249 | 3.94 | 1.6 |
Urine +50 % | 308 | 3.71 | 1.2 |
Qualitative precision - 200 ng/mL cutoff | |||
Cutoff (200) | Number | Correct | Confidence level |
Urine -50 % | 84 | 84 | > 95 % negative reading |
DAT3N | 84 | 84 | > 95 % negative reading |
Cutoff urine | 84 | n.a.* | n.a.* |
DAT3P | 84 | 84 | > 95 % positive reading |
Urine +50 % | 84 | 84 | > 95 % positive reading |
*n.a. = not applicable
Semiquantitative precision - 300 ng/mL cutoff | |||
Repeatability | Mean | SD | CV |
Urine -50 % | 150 | 7.03 | 4.7 |
DAT1N | 231 | 8.04 | 3.5 |
Cutoff urine | 345 | 7.59 | 2.2 |
DAT1P | 375 | 7.66 | 2.0 |
Urine +50 % | 435 | 8.89 | 2.0 |
Intermediate | Mean | SD | CV |
Urine -50 % | 150 | 10.5 | 7.0 |
DAT1N | 231 | 10.0 | 4.3 |
Cutoff urine | 345 | 10.6 | 3.1 |
DAT1P | 375 | 9.26 | 2.5 |
Urine +50 % | 435 | 11.7 | 2.7 |
Qualitative precision - 300 ng/mL cutoff | |||
Cutoff (300) | Number | Correct | Confidence level |
Urine -50 % | 84 | 84 | > 95 % negative reading |
DAT1N | 84 | 84 | > 95 % negative reading |
Cutoff urine | 84 | n.a.* | n.a.* |
DAT1P | 84 | 84 | > 95 % positive reading |
Urine +50 % | 84 | 84 | > 95 % positive reading |
*n.a. = not applicable
The benzodiazepines constitute a class of versatile and widely prescribed central nervous system (CNS) depressant drugs with medically useful anxiolytic, sedative, hypnotic, muscle relaxant, and anticonvulsant activities.
Following ingestion, the benzodiazepines of the 1,4‑substituted class (including the triazolobenzodiazepine derivatives) are absorbed, metabolized, and excreted in the urine at different rates as a variety of structurally related metabolites. Metabolite diversity reflects the different physiochemical properties and metabolic pathways of the individual drugs. Overall metabolic similarities include removal of substituents from the β ring of the 1,4‑substituted benzodiazepines, α‑hydroxylation of the triazolobenzodiazepines, demethylation, hydroxylation of the three‑position carbon of the β ring, and conjugation of hydroxylated metabolites followed by urinary excretion predominantly as glucuronides.
R1 | Benzodiazepines antibody (sheep polyclonal); buffer; β‑glucuronidase enzyme; bovine serum albumin (BSA); 0.09 % sodium azide |
R2 | Conjugated benzodiazepine derivative microparticles; buffer; 0.09 % sodium azide |
R1 is in position B and R2 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.
For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.
Drug concentrations of Control Set DAT I, II, III and Clinical have been verified by GC‑MS.
The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 26 weeks.
Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
Only the specimens listed below were tested and found acceptable.
Urine: Collect urine samples in clean glass or plastic containers. Fresh urine specimens do not require any special handling or pretreatment, but an effort should be made to keep pipetted samples free of gross debris. Samples should be within the normal physiological pH range of 5‑8. No additives or preservatives are required. It is recommended that urine specimens be stored at 2‑8 °C and tested within 5 days of collection.
For prolonged storage, freezing of samples is recommended.
Centrifuge highly turbid specimens before testing.
See the limitations and interferences section for details about possible sample interferences.
Adulteration or dilution of the sample can cause erroneous results. If adulteration is suspected, another sample should be collected. Specimen validity testing is required for specimens collected under the Mandatory Guidelines for Federal Workplace Drug Testing Programs.
CAUTION: Specimen dilutions should only be used to interpret results of Calc.? and Samp.? alarms, or when estimating concentration in preparation for GC‑MS or LC‑MS/MS. Dilution results are not intended for patient values. Dilution procedures, when used, should be validated.
ONLINE DAT Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative detection of benzodiazepines in human serum and plasma on Roche/Hitachi cobas c systems at a cutoff concentration of 200 ng/mL.
Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC‑MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC‑MS/MS) is the preferred confirmatory method.
The assay is based on the kinetic interaction of microparticles in a solution (KIMS)
When a serum sample contains the drug in question, this drug competes with the particle‑bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.
The presence of β‑glucuronidase enzyme enhances the Benzodiazepines II assay cross‑reactivity to some of the glucuronidated metabolites.
Qualitative assay
Results of this assay distinguish preliminary positive (≥ 200 ng/mL) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.
Criterion: No cross‑over at initial values of samples of 100 ng/mL and 300 ng/mL (control levels).
See the \"Specific performance data\" section of this document for information on substances tested with this assay. There is the possibility that other substances and/or factors may interfere with the test and cause erroneous results (e.g., technical or procedural errors).
A preliminary positive result with this assay indicates the presence of benzodiazepines and/or their metabolites in serum. It does not reflect the degree of intoxication.
Icterus:
Hemolysis:
Lipemia (Intralipid):
Rheumatoid factors: No significant interference from rheumatoid factors up to a concentration of 1200 IU/mL.
Immunoglobulins: No significant interference from immunoglobulins up to a concentration of 16 g/L (simulated by human immunoglobulin A), up to a concentration of 70 g/L (simulated by human immunoglobulin G) and up to a concentration of 10 g/L (simulated by human immunoglobulin M).
Albumin: No significant interference from human serum albumin up to a concentration of 70 g/L.
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
|
| Analyzer(s) on which cobas c pack(s) can be used | |
08056927190 | ONLINE DAT Benzodiazepines II (850 tests) | System‑ID 2028 001 | cobas c 503 |
Materials required (but not provided):
03304671190 | Preciset DAT Plus I calibrator CAL 5 | Code 20435 | |
07978766190 | Serum DAT Control Low (ACQ Partner Channel*) | ||
07978740190 | Serum DAT Control High (ACQ Partner Channel*) | ||
08063494190 | NaCl Diluent 9 % (123 mL) | System‑ID 2906 001 |
*Roche does not hold the product registration for Partner Channels. The legal manufacturer indicated on the kit is solely responsible for all of the design, legal, and regulatory aspects of the product.
BEQ2S: ACN 20287 (Serum/plasma): for qualitative assay, 200 ng/mL
Ready for use
Carefully invert reagent container several times prior to use to ensure that the reagent components are mixed.
Test definition | |
Qualitative | |
Reporting time | 10 min |
Wavelength (sub/main) | – /546 nm |
Reagent pipetting | |
R1 | 63 µL |
R2 | 28 µL |
Sample volumes | Sample |
200 ng/mL cutoff | |
Normal | 3.2 µL |
Decreased | 3.2 µL |
Increased | 3.2 µL |
For further information about the assay test definitions refer to the application parameters setting screen of the corresponding analyzer and assay.
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 26 weeks |
Do not freeze. |
Calibrators | Qualitative application |
200 ng/mL cutoff assay | |
S1: Preciset DAT Plus I calibrator ‑ CAL 5, 1000 ng/mL with automatic pre‑dilution | |
The drug concentration of the calibrator has been verified by GC‑MS. | |
Calibration K factor | For the qualitative application a K factor of -1000 is predefined in the application settings. |
Calibration mode | Qualitative application |
Linear | |
Calibration frequency | Full calibration |
For the cutoff calibrator a value of \"0\" is encoded in the e‑barcode in order to ensure flagging of positive samples with >Test and negative absorbance values for negative samples.
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against a primary reference method (GC‑MS).
Representative performance data on the analyzers are given below. These data represent the performance of the analytical procedure itself.
Results obtained in individual laboratories may differ due to heterogenous sample materials, aging of analyzer components and mixture of reagents running on the analyzer.
Precision was determined using human samples and controls in accordance with the CLSI (Clinical and Laboratory Standards Institute) EP05‑A3 requirements with repeatability (n = 84) and intermediate precision (2 aliquots per run, 2 runs per day, 21 days). The following results were obtained:
Cutoff (200) | Number | Correct | Confidence level |
Serum -75 % | 84 | 84 | > 95 % negative reading |
ACQ‑L | 84 | 84 | > 95 % negative reading |
Cutoff serum | 84 | n.a.** | n.a.** |
ACQ‑H | 84 | 84 | > 95 % positive reading |
Serum +75 % | 84 | 84 | > 95 % positive reading |
**n.a. = not applicable
The benzodiazepines constitute a class of versatile and widely prescribed central nervous system (CNS) depressant drugs with medically useful anxiolytic, sedative, hypnotic, muscle relaxant, and anticonvulsant activities.
Following ingestion, the benzodiazepines of the 1,4‑substituted class (including the triazolobenzodiazepine derivatives) are absorbed, metabolized, and excreted in the urine at different rates as a variety of structurally related metabolites. Metabolite diversity reflects the different physiochemical properties and metabolic pathways of the individual drugs. Overall metabolic similarities include removal of substituents from the β ring of the 1,4‑substituted benzodiazepines, α‑hydroxylation of the triazolobenzodiazepines, demethylation, hydroxylation of the three‑position carbon of the β ring, and conjugation of hydroxylated metabolites followed by urinary excretion predominantly as glucuronides.
R1 | Benzodiazepines antibody (sheep polyclonal); buffer; β‑glucuronidase enzyme; bovine serum albumin (BSA); 0.09 % sodium azide |
R2 | Conjugated benzodiazepine derivative microparticles; buffer; 0.09 % sodium azide |
R1 is in position B and R2 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.
For quality control, use control materials as listed in the “Order information” section. In addition, other suitable control material can be used.
Drug concentrations of the high and low controls have been verified by GC‑MS.
The control intervals and limits should be adapted to each laboratory’s individual requirements. It is recommended to perform quality control always after lot calibration and subsequently at least every 26 weeks.
Values obtained should fall within the defined limits. Each laboratory should establish corrective measures to be taken if values fall outside the defined limits.
Follow the applicable government regulations and local guidelines for quality control.
For specimen collection and preparation only use suitable tubes or collection containers.
Only the specimens listed below were tested and found acceptable.
Serum: Serum tubes with and without separating gel.
Plasma: K2‑ or K3‑EDTA, lithium heparin plasma.
Stability: | 5 days capped at 15‑25 °C |
14 days capped at 2‑8 °C | |
6 months capped at -20 °C |
The sample types listed were tested with a selection of sample collection tubes that were commercially available at the time of testing, i.e. not all available tubes of all manufacturers were tested. Sample collection systems from various manufacturers may contain differing materials which could affect the test results in some cases. When processing samples in primary tubes (sample collection systems), follow the instructions of the tube manufacturer.
Centrifuge samples containing precipitates before performing the assay.
See the limitations and interferences section for details about possible sample interferences.
Specimens can be repeatedly frozen and thawed up to 3 times.
Invert thawed specimens several times prior to testing.
CAUTION: Specimen dilutions should only be used to interpret results of Calc.? and Samp.? alarms, or when estimating concentration in preparation for GC‑MS or LC‑MS/MS. Dilution results are not intended for patient values. Dilution procedures, when used, should be validated.
Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on Roche/Hitachi cobas c systems at cutoff concentrations of 100 ng/mL, 200 ng/mL, and 300 ng/mL.
Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program.
Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC‑MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC‑MS/MS) is the preferred confirmatory method.
The assay is based on the kinetic interaction of microparticles in a solution (KIMS)
When a urine sample contains the drug in question, this drug competes with the particle‑bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.
The presence of β‑glucuronidase enzyme enhances the Benzodiazepines II assay cross‑reactivity to some of the glucuronidated metabolites.
Qualitative assay
Results of this assay distinguish preliminary positive (≥ 100 ng/mL, ≥ 200 ng/mL, or ≥ 300 ng/mL depending on the cutoff) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.
Semiquantitative assay
Results of this assay yield only approximate cumulative concentrations of the drug and its metabolites (see Analytical specificity section).
See the \"Specific performance data\" section of this document for information on substances tested with this assay. There is the possibility that other substances and/or factors may interfere with the test and cause erroneous results (e.g., technical or procedural errors).
A preliminary positive result with this assay indicates the presence of benzodiazepines
and/or their metabolites in urine. It does not reflect the degree of intoxication.
Interfering substances were added to urine containing nordiazepam at ‑25 % and +25 % of the cutoff level at the concentration listed below. Samples were tested and the following results were obtained on a Roche/Hitachi 917 analyzer.
Semiquantitative (ng/mL) | 100 ng/mL Cutoff | 200 ng/mL Cutoff | 300 ng/mL Cutoff | ||||
Compound | Cmpd. Conc. | Neg Level | Pos Level | Neg Level | Pos Level | Neg Level | Pos Level |
Acetone | 1 | 77(NEG) | 134(POS) | 157(NEG) | 260(POS) | 231(NEG) | 402(POS) |
Ascorbic acid | 1.5 | 78(NEG) | 132(POS) | 156(NEG) | 262(POS) | 233(NEG) | 399(POS) |
Conjugated bilirubin | 0.25 | 82(NEG) | 129(POS) | 156(NEG) | 247(POS) | 229(NEG) | 392(POS) |
Creatinine | 5 | 81(NEG) | 138(POS) | 158(NEG) | 259(POS) | 230(NEG) | 396(POS) |
Ethanol | 1 | 78(NEG) | 136(POS) | 151(NEG) | 261(POS) | 228(NEG) | 395(POS) |
Glucose | 20 | 81(NEG) | 138(POS) | 158(NEG) | 262(POS) | 236(NEG) | 403(POS) |
Hemoglobin | 1 | 76(NEG) | 139(POS) | 159(NEG) | 261(POS) | 228(NEG) | 398(POS) |
Human serum albumin | 5 | 83(NEG) | 140(POS) | 165(NEG) | 273(POS) | 243(NEG) | 422(POS) |
Oxalic acid | 2 | 74(NEG) | 128(POS) | 151(NEG) | 254(POS) | 226(NEG) | 388(POS) |
Sodium chloride | 0.5 | 79(NEG) | 139(POS) | 159(NEG) | 262(POS) | 234(NEG) | 389(POS) |
Urea | 6 | 80(NEG) | 138(POS) | 157(NEG) | 261(POS) | 233(NEG) | 405(POS) |
The same experiment was performed in the qualitative mode for each cutoff. All negative and positive samples recovered properly in the presence of the interfering substance.
An additional protocol was executed in which samples containing nordiazepam at control levels (±25 % of cutoff) with specific gravities ranging from 1.006 to 1.034 were tested. As with the other interferences, there were no control cross‑overs on any of the 3 assay cutoffs at either extreme specific gravity level.
For diagnostic purposes, the results should always be assessed in conjunction with the patient’s medical history, clinical examination and other findings.
ACTION REQUIRED
Special Wash Programming: The use of special wash steps is mandatory when certain test combinations are run together on cobas c systems. All special wash programming necessary for avoiding carry‑over is available via the cobas link, manual input is required in certain cases. The latest version of the carry‑over evasion list can be found with the NaOHD/SMS/SmpCln1+2/SCCS Method Sheet and for further instructions refer to the operator’s manual.
Where required, special wash/carry‑over evasion programming must be implemented prior to reporting results with this test.
|
| Analyzer(s) on which cobas c pack(s) can be used | |
06334415190 | ONLINE DAT Benzodiazepines II 100 tests | System‑ID 01 6997 5 |
Materials required (but not provided):
03304671190 | Preciset DAT Plus I calibrators CAL 1‑6 (6 x 5 mL) | Codes 431‑436 | |
03304680190 | Preciset DAT Plus II calibrators CAL 1‑6 (6 x 5 mL) | Codes 437‑442 | |
03304698190 | C.f.a.s. DAT Qualitative Plus (6 x 5 mL) | Code 433 | |
04590856190 | C.f.a.s. DAT Qualitative Plus Clinical ( 3 x 5 mL) | Code 699 | |
03312950190 | Control Set DAT I (for 300 ng/mL assay) | ||
03312968190 | Control Set DAT II (for 100 ng/mL assay) | ||
04500873190 | Control Set DAT Clinical (for 100 ng/mL assay) | ||
03312976190 | Control Set DAT III (for 200 ng/mL assay) |
BZ1Q2: ACN 8718: for qualitative assay, 100 ng/mL
BZ2Q2: ACN 8719: for qualitative assay, 200 ng/mL
BZ3Q2: ACN 8720: for qualitative assay, 300 ng/mL
BZ1S2: ACN 8728: for semiquantitative assay, 100 ng/mL
BZ2S2: ACN 8729: for semiquantitative assay, 200 ng/mL
BZ3S2: ACN 8730: for semiquantitative assay, 300 ng/mL
BZQ1C: ACN 8727: for qualitative assay, 100 ng/mL;using C.f.a.s. DAT Qualitative Plus Clinical
Ready for use
Carefully invert reagent container several times prior to use to ensure that the reagent components are mixed.
Deselect Automatic Rerun for these applications in the Utility menu, Application screen, Range tab.
cobas c 701/702 test definition | |||
Semiquantitative | Qualitative | ||
Assay type | 2‑Point End | 2‑Point End | |
Reaction time / Assay points | 10 / 10‑26 | 10 / 10‑26 | |
Wavelength (sub/main) | – /546 nm | – /546 nm | |
Reaction direction | Increase | Increase | |
Unit | ng/mL | mAbs | |
Reagent pipetting | Diluent (H2O) | ||
R1 | 90 µL | – | |
R2 | 40 µL | – | |
Sample volumes | Sample | Sample dilution | |
100 and 200 ng/mL cutoffs | Sample | Diluent (NaCl) | |
Normal | 4.5 µL | – | – |
Decreased | 4.5 µL | – | – |
Increased | 4.5 µL | – | – |
300 ng/mL cutoff | |||
Normal | 2.0 µL | – | – |
Decreased | 2.0 µL | – | – |
Increased | 2.0 µL | – | – |
Shelf life at 2‑8 °C: | See expiration date on cobas c pack label |
On‑board in use and refrigerated on the analyzer: | 8 weeks |
On‑board on the Reagent Manager: | 24 hours |
Do not freeze. |
Calibrators | Semiquantitative applications |
100 and 200 ng/mL cutoff assays | |
S1‑6: Preciset DAT Plus II calibrators, CAL 1‑6 0, 50, 100, 200, 400, 1000 ng/mL | |
300 ng/mL cutoff assay | |
S1‑6: Preciset DAT Plus I calibrators, CAL 1‑6 0, 150, 300, 600, 1000, 3000 ng/mL | |
Qualitative applications | |
100 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 3 (Test BZ1Q2) , 100 ng/mL, S1: C.f.a.s. DAT Qualitative Plus Clinical (Test BZQ1C) , 100 ng/mL | |
200 ng/mL cutoff assay | |
S1: Preciset DAT Plus II calibrator - CAL 4, 200 ng/mL | |
300 ng/mL cutoff assay | |
S1: C.f.a.s. DAT Qualitative Plus or Preciset DAT Plus I calibrator - CAL 3, 300 ng/mL | |
The drug concentrations of the calibrators have been verified by GC‑MS. | |
Calibration K Factor | For the qualitative applications, enter the K Factor as -1000 into the Calibration menu, Status screen, Calibration Result window. |
Calibration mode | Semiquantitative applications |
Result Calculation Mode (RCM) FREFSee Results section | |
Qualitative applications | |
Linear | |
Calibration frequency | Full (semiquantitative) or blank (qualitative) calibration |
Calibration interval may be extended based on acceptable verification of calibration by the laboratory.
Traceability: This method has been standardized against a primary reference method (GC‑MS).
Representative performance data on the analyzers are given below. Results obtained in individual laboratories may differ.
Precision was determined in an internal protocol by running a series of calibrator and controls with repeatability (n = 21) and intermediate precision (n = 40). The following results were obtained:
100 ng/mL cutoff | |||
Semiquantitative precision | |||
Repeatability | Mean ng/mL | SD ng/mL | CV % |
Level 1 | 80 | 3 | 3.6 |
Level 2 | 98 | 3 | 2.9 |
Level 3 | 129 | 3 | 2.0 |
Intermediate precision | Mean ng/mL | SD ng/mL | CV % |
Level 1 | 78 | 3 | 3.7 |
Level 2 | 101 | 2 | 2.1 |
Level 3 | 124 | 4 | 3.1 |
Qualitative precision | |||
Cutoff (100) | Number tested | Correct results | Confidence level |
0.75x | 105 | 105 | > 95 % negative reading |
1.25x | 105 | 105 | > 95 % positive reading |
200 ng/mL cutoff | |||
Semiquantitative precision | |||
Repeatability | Mean ng/mL | SD ng/mL | CV % |
Level 1 | 149 | 3 | 1.8 |
Level 2 | 192 | 3 | 1.4 |
Level 3 | 247 | 3 | 1.3 |
Intermediate precision | Mean ng/mL | SD ng/mL | CV % |
Level 1 | 145 | 5 | 3.7 |
Level 2 | 191 | 5 | 2.4 |
Level 3 | 241 | 8 | 3.1 |
Qualitative precision | |||
Cutoff (200) | Number tested | Correct results | Confidence level |
0.75x | 105 | 105 | > 95 % negative reading |
1.25x | 105 | 105 | > 95 % positive reading |
300 ng/mL cutoff | |||
Semiquantitative precision | |||
Repeatability | Mean ng/mL | SD ng/mL | CV % |
Level 1 | 227 | 7 | 3.1 |
Level 2 | 297 | 8 | 2.6 |
Level 3 | 386 | 7 | 1.7 |
Intermediate precision | Mean ng/mL | SD ng/mL | CV % |
Level 1 | 231 | 6 | 2.7 |
Level 2 | 301 | 7 | 2.3 |
Level 3 | 364 | 8 | 2.3 |
Qualitative precision | |||
Cutoff (300) | Number tested | Correct results | Confidence level |
0.75x | 105 | 105 | > 95 % negative reading |
1.25x | 105 | 105 | > 95 % positive reading |
Results for intermediate precision were obtained on the master system cobas c 501 analyzer.
The benzodiazepines constitute a class of versatile and widely prescribed central nervous system (CNS) depressant drugs with medically useful anxiolytic, sedative, hypnotic, muscle relaxant, and anticonvulsant activities.
Following ingestion, the benzodiazepines of the 1,4‑substituted class (including the triazolobenzodiazepine derivatives) are absorbed, metabolized, and excreted in the urine at different rates as a variety of structurally related metabolites. Metabolite diversity reflects the different physiochemical properties and metabolic pathways of the individual drugs. Overall metabolic similarities include removal of substituents from the β ring of the 1,4‑substituted benzodiazepines, α-hydroxylation of the triazolobenzodiazepines, demethylation, hydroxylation of the three-position carbon of the β ring, and conjugation of hydroxylated metabolites followed by urinary excretion predominantly as glucuronides.
R1 | Benzodiazepines antibody (sheep polyclonal); buffer; β‑glucuronidase enzyme; bovine serum albumin (BSA); 0.09 % sodium azide |
R2 | Conjugated benzodiazepine derivative microparticles; buffer; 0.09 % sodium azide |
R1 is in position B and R2 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.
For quality control, use control materials as listed in the \"Order information\" section.
In addition, other suitable control material can be used.
Drug concentrations of the controls have been verified by GC‑MS.
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.
Only the specimens listed below were tested and found acceptable.
Urine: Collect urine samples in clean glass or plastic containers. Fresh urine specimens do not require any special handling or pretreatment, but an effort should be made to keep pipetted samples free of gross debris. Samples should be within the normal physiological pH range of 5‑8. No additives or preservatives are required. It is recommended that urine specimens be stored at 2‑8 °C and tested within 5 days of collection.
For prolonged storage, freezing of the sample is recommended.
Centrifuge highly turbid specimens before testing.
Adulteration or dilution of the sample can cause erroneous results. If adulteration is suspected, another sample should be collected. Specimen validity testing is required for specimens collected under the Mandatory Guidelines for Federal Workplace Drug Testing Programs.
Specimens containing human‑sourced materials should be handled as if potentially infectious using safe laboratory procedures such as those outlined in Biosafety in Microbiological and Biomedical Laboratories (HHS Publication Number [CDC] 93‑8395).
CAUTION: Specimen dilutions should only be used to interpret results of Calc.? and Samp.? alarms, or when estimating concentration in preparation for GC‑MS or LC‑MS/MS. Dilution results are not intended for patient values. Dilution procedures, when used, should be validated.
ONLINE DAT Benzodiazepines II
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