AMPHETAMINES II ASSAY
Device Facts
| Record ID | K093664 |
|---|---|
| Device Name | AMPHETAMINES II ASSAY |
| Applicant | Roche Diagnostics Corp. |
| Product Code | DKZ · Clinical Toxicology |
| Decision Date | Jul 28, 2010 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 862.3100 |
| Device Class | Class 2 |
Intended Use
Amphetamines II (AMPII) is an in vitro diagnostic test for the qualitative and semiquantitative detection of amphetamines and methamphetamines on COBAS INTEGRA systems at cutoff concentrations of 300 ng/mL, 500 ng/mL and 1000 ng/mL when calibrated with d-methamphetamine. 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). Amphetamines 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) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
Device Story
Immunoassay for automated clinical chemistry analyzers (Integra 800); detects amphetamines/methamphetamines in urine. Principle: Kinetic interaction of microparticles in solution (KIMS). Input: Urine sample; antibody working solution; drug-microparticle conjugate. Mechanism: In absence of drug, antibody binds microparticles causing aggregation (absorbance increases). If drug present, it competes for antibody, inhibiting aggregation; absorbance change proportional to drug concentration. Output: Preliminary positive/negative result (qualitative) or concentration (semi-quantitative). Used by laboratory professionals to screen for drug abuse; results require GC/MS confirmation. Benefits: Rapid preliminary screening for clinical decision-making.
Clinical Evidence
Bench testing only. Precision was evaluated per CLSI EPS-A2 (N=84 per condition). Accuracy was determined by comparing results against GC/MS-confirmed urine samples. Qualitative and semiquantitative agreement with GC/MS was reported, with 100% agreement for high-positive samples and varying agreement for near-cutoff samples. Analytical specificity and cross-reactivity were assessed against various phenethylamines and unrelated drugs. No clinical trial data was required.
Technological Characteristics
Wet reagent immunoassay (KIMS). Reagents: Antibody working solution (mouse monoclonal) and drug-microparticle conjugate in buffer with BSA and 0.09% sodium azide. Platform: COBAS INTEGRA 800. Calibration: d-methamphetamine. Stability: 1 year closed vial (2-8°C), 56 days on-board. Connectivity: Automated analyzer.
Indications for Use
Indicated for qualitative and semiquantitative detection of amphetamines and methamphetamines in human urine. Used for preliminary screening; requires confirmation by GC/MS. Intended for professional use in clinical laboratories.
Regulatory Classification
Identification
An amphetamine test system is a device intended to measure amphetamine, a central nervous system stimulating drug, in plasma and urine. Measurements obtained by this device are used in the diagnosis and treatment of amphetamine use or overdose and in monitoring levels of amphetamine to ensure appropriate therapy.
Special Controls
*Classification.* Class II (special controls). An amphetamine test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (*e.g.,* programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).
Predicate Devices
- ONLINE Amphetamines II Assay (k083764)
Reference Devices
- Preciset DAT Plus I Calibrators (K060645)
- Preciset DAT Plus II Calibrators (K060645)
- C.f.a.s. DAT Qualitative Plus Calibrators (K060645)
- Control Set DAT I (K080183)
- Control Set DAT II (K080183)
- Control Set DAT III (K080183)
Related Devices
- K083764 — AMPHETAMINES II · Roche Diagnostics Corp. · Feb 3, 2010
- K983699 — MODIFICATION TO ABUSCREEN ONLINE FOR AMPHETAMINES · Roche Diagnostic Systems, Inc. · Dec 11, 1998
- K110446 — ROCHE ORAL FLUID AMPHETAMINE, ROCHE ORAL FLUID DAT QUAL CAL B, ROCHE ORAL FLUID DAT CONTROL SET B, AND ROCHE ORAL FLUID · Roche Diagnostics · May 3, 2011
Submission Summary (Full Text)
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## 510(k) Summary: Amphetamines II Assay for Integra Family of Analyzers
According to the requirements of 21 CFR 807.92, the following Introduction information provides sufficient detail to understand the basis for a determination of substantial equivalence.
Submitter Roche Diagnostics Name, Address, 9115 Hague Rd. Contact Indianapolis, IN 46250 317-521-3742
JUL 2 8 2010
Contact Person: Michelle Neff
Kogzblogy
Proprietary name: Amphetamines II Device Name ·
Common name: Amphetamine metabolite test system
Classification name: Enzyme Immunoassay, Amphetamine
Product Code: DKZ
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#### Device Description
The Amphetamines II test is an immunoassay for use on automated clinical chemistry analyzers. The device consists of two wet reagents; a soluble drug-conjugate, and an antibody-bound microparticle solution. During the assay, in the absence of sample drug in urine, soluble drug-conjugates bind to antibody-bound microparticles, causing the formation of particle aggregates. When a urine sample contains the drug in question, this drug competes with the drug derivative conjugate for microparticle-bound antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The rate of absorbance change is proportional to the concentration of drug in the sample. Calibrators, ranging in concentration from 0-5000 ng/mL depending on cutoff and test mode, are run with the assay. Concentrations of controls and unknowns are calculated from the standard curve in semi-quantitative mode. Results for controls or calibrators are determined as preliminary positive or negative relative to the cutoff in qualitative mode.
C.f.a.s. DAT Qualitative Clinical, C.f.a.s. DAT Qualitative Plus, C.f.a.s. DAT Qualitative Plus Clinical, Preciset DAT Plus I Calibrators, and Preciset DAT Plus II Calibrators are ready to use, multianalyte calibrators prepared by the quantitative addition of drug or drug metabolite to drug-free human urine.
Control Set DAT I, II, and III, and Control Set DAT Clinical are ready to use multianalyte controls prepared by the quantitative addition of drug or drug metabolite to drug-free urine.
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Amphetamines II (AMPII) is an in vitro diagnostic test for the qualitative and semiquantitative detection of amphetamines and methamphetamines on COBAS INTEGRA systems in human urine at cutoff concentrations of 300 ng/mL, 500 ng/mL and 1000 ng/mL when calibrated with d-methamphetamine. 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).
Amphetamines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in obtain a a confirmed a analytical aresult. Gas order to chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method.1 Clinical consideration and professional judgement should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
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#### Comparison to the Predicate Device
The ONLINE Amphetamines II assay is substantially equivalent to other products in commercial distribution intended for similar use. Most notably, we claim substantial equivalence to the currently marketed Amphetamines II assay (K083764).
The ONLINE Amphetamines II assay contains an in vitro diagnostic reagent system intended for use on the Roche Integra analyzers for the semi-quantitative and qualitative detection of amphetamines in human urine at cutoff concentrations of 300, 500, and 1000 ng/mL.
The Roche ONLINE Amphetamines II assay utilizes a modified KIMS technology relative to the currently marketed Abuscreen OnLine Amphetamines assay. Differences between this application and the cleared assay include:
- 1. use of amphetamine and methamphetamine, and monoclonal antibodies attached to microparticles in solution
- 2. a soluble drug-polymer conjugate
- 3. increased sensitivity to "designer drugs"and their metabolites, and
- 4. addition of 300 and 500 ng/mL cutoff concentrations.
The recommended calibrators to be used with the proposed ONLINE Amphetamines II assay are the Preciset DAT Plus I, Preciset DAT Plus II and Cfas DAT Qualitative Plus Calibrators (K060645).
The recommended controls to be used with the proposed ONLINE Amphetamines II assay are the Control Set DAT I, Control Set DAT II, Control Set DAT III (K080183).
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| Comparison of Technological Characteristics | | | | |
|---------------------------------------------|---------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|--|
| Feature | Amphetamines II<br>Assay, Integra 800 | Predicate Device: Amphetamines II<br>Assay, Hitachi 917 (K083764) | | |
| Methodology | Same | KIMS, Kinetic interaction of<br>microparticles in solution | | |
| Sample Type | Same | Urine | | |
| Intended Use | Same | Qualitative and semi-quantitative<br>detection of amphetamine and<br>methamphetamine | | |
| Reagent | Same | 1. Conjugate Working Solution: Conjugated<br>amphetamine and methamphetamine<br>derivatives in buffer with bovine serum<br>albumin(BSA) and 0.09% sodium azide.<br>2. Antibody/Microparticle Working Solution:<br>Microparticles attached to amphetamine and<br>methamphetamine antibodies (mouse<br>monoclonal) in buffer with bovine serum<br>albumin (BSA) and 0.09% sodium azide. | | |
| Cutoff | Same | 300, 500, 1000 ng/mL | | |
| Calibrator/Control<br>Matrix | Same | Human urine | | |
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# AMPII
Amphetamines II
| Order information | | |
|--------------------------|-----------|-----------------------|
| COBAS INTEGRA | 200 Tests | Cat. No. 04512936 190 |
| ONLINE DAT | | System-ID 07 6835 9 |
| Amphetamines II | | |
| Preciset DAT Plus I | 6 × 5 mL | Cat. No. 03304671 190 |
| calibrators | | |
| CAL 1-6 | | |
| Preciset DAT Plus II | 6 × 5 mL | Cat. No. 03304680 190 |
| calibrators | | |
| CAL 1-6 | | |
| C.f.a.s. DAT Qualitative | 6 x 5 mL | Cat. No. 03304698 190 |
| Plus | | |
| C.f.a.s. DAT Qualitative | 3 x 5 mL | Cat. No. 04590856 190 |
| Plus Clinical | | |
| Control Set DAT II | | Cat. No. 03312968 190 |
| (for 300 ng/mL assay) | | |
| PreciPos DAT Set II | 2 x 10 mL | |
| PreciNeg DAT Set II | 2 x 10 mL | |
| Control Set DAT I | | Cat. No. 03312950 190 |
| (for 500 ng/mL assay) | | |
| PreciPos DAT Set I | 2 x 10 mL | |
| PreciNeg DAT Set I | 2 x 10 mL | |
| Control Set DAT Clinical | | Cat. No. 04500873 190 |
| (for 500 ng/mL assay) | | |
| PreciPos DAT Clinical | 2 x 10 mL | |
| PreciNeg DAT Clinical | 2 x 10 mL | |
| Control Set DAT III | | Cat. No. 03312976 190 |
| (for 1000 ng/mL assay) | | |
| PreciPos DAT Set III | 2 x 10 mL | |
| PreciNeg DAT Set III | 2 x 10 mL | |
| | | |
#### System information
Test AM3S2: 0-358: for semiquantitative assay, 300 ng/mL Test AM5S2: 0-359: for semiquantitative assay, 500 ng/mL Test AM1S2: 0-360: for semiquantitative assay, 1000 ng/mL Test AM3Q2: 0-330: for qualitative assay, 300 ng/mL Test AM5Q2: 0-340: for qualitative assay, 500 ng/mL Test AM1Q2: 0-350: for qualitative assay, 1000 ng/mL Test AM5QC: 0-341: for qualitative assay, 500 ng/mL; using C.f.a.s. DAT Qualitative Plus Clinical
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#### Intended use
Amphetamines II (AMPII) is an in vitro diagnostic test for the qualitative and semiquantitative detection of amphetamines and methamphetamines in human urine on COBAS INTEGRA systems at cutoff concentrations of 300 ng/mL, 500 ng/mL and 1000 ng/mL when calibrated with d-methamphetamine. 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).
Amphetamines 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. GC/MS is the preferred confirmatory method.1 Clinical consideration and professional judgement should be applied to any drug of abuse test result, particularly when preliminary positive results are used.
#### Summarv
The amphetamines are known as the sympathomimetic amines as they mimic the effects of stimulation of the sympathetic nervous system. These small molecules, based on Bphenvlethylamine, structurally resemble the bodies own catecholamines. A wide variety have been created via substitutions anywhere on the structure. The amphetamines are potent central nervous stimulants. As such they can increase wakefulness, physical activity, and decrease appetite. The amphetamines have some limited indications and approval for use in ADHD, narcolepsy, and obesity. However, because these CNS stimulants convey a sense of self-confidence, well being, and euphoria, they are highly addictive, widely abused, and consequently controlled substances.2 Abuse can lead to medical, psychological, and social consequences. Adverse health effects include memory loss, aggression, psychotic behavior, heart damage, malnutrition, and severe dental problems.2 Amphetamine may be self-administered either orally or by intravenous injection in amounts of up to 2000 mg daily by tolerant addicts. It is a metabolite of a number of other drugs including methamphetamine. Normally about 30 % is excreted unchanged in the 24 hour urine, but this may change to as much as 74 % in acid urine and may decrease to 1 % in alkaline urine.4 Amphetamines II is calibrated with d-methamphetamine and therefore the sensitivity towards amphetamines is different than d-methamphetamine, as indicated in the "Analytical specificity" section.
#### Test principle
The assay is based on the kinetic interaction of microparticles in a solution (KIMS) 5 as measured by changes in light transmission. In the absence of amphetamine or methamphetamine, soluble drug conjugates bind to antibody-bound microparticles, causing the formation of particle aggregates. As the aggregation reaction proceeds in the absence of amphetamine or methamphetamine, the absorbance increases.
When a urine sample contains amphetamine or methamphetamine, this drug competes with the drug derivative conjugate for microparticle-bound antibody. Antibody bound to amphetamine or methamphetamine is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of amphetamine diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Amphetamine or methamphetamine content is determined relative to the value obtained for a known cutoff concentration of d-methamphetamine.7
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#### Reagents - working solutions
- Conjugated amphetamine and methamphetamine derivatives; buffer; bovine serum R1 albumin: 0.09 % sodium azide
- Microparticles attached to amphetamine and methamphetamine antibodies (mouse SR monoclonal); buffer; bovine serum albumin; 0.09 % sodium azide
#### Precautions and warnings
Pay attention to all precautions and warnings listed in this Method Manual, Chapter 1, Introduction.
#### Reagent handling
COBAS INTEGRA 800 analyzers
The reagent is automatically mixed for 1 minute after cobas c pack puncture and for half a minute during Begin of Day.
#### Storage and stability
Shelf life at 2 to 8 °C
See expiration date on cobas c pack label
### COBAS INTEGRA 800 analyzers
On-board in use at 8 °C
84 days
The on-board in use stability period begins at the time of cobas c pack puncture. Do not freeze reagents. Reagents that have been frozen should be discarded.
#### Specimen collection and preparation
Only the specimens listed below were tested and found acceptable.
Urine: Collect urine samples in clean glass or plastic containers. Fresh urine samples do not require 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 to 8. No additives or preservatives are required. It is recommended that urine specimens be stored at 2 to 8 °C and tested within 5 days of collection . 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.2 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 as an estimation for GC/MS and are not intended for patient values. Dilution procedures, when used, should be validated.
#### Materials provided
See "Reagents - working solutions" section for reagents.
#### Assay
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For optimum performance of the assay follow the directions given in this document for the analyzer concerned. Refer to the appropriate operator's manual for analyzer-specific assay instructions.
### Application for urine
### COBAS INTEGRA 800 test definition
| | Semiquantitative | Qualitative |
|-----------------------------------------------------------------------------|------------------|---------------|
| Measuring mode | Absorbance | Absorbance |
| Abs. calculation mode | Endpoint | Endpoint |
| Reaction mode | R1-S-SR | R1-S-SR |
| Reaction direction | Increase | Increase |
| Reaction start | SR | SR |
| Wavelength A | 659 nm | 659 nm |
| Test range | | |
| <i>AM3S2</i> | 0-2000 ng/mL | 0-2000 |
| <i>AM5S2, AM1S2</i> | 0-5000 ng/mL | |
| with postdilution | | |
| <i>AM3S2</i> | 0-20000 ng/mL | |
| <i>AM5S2, AM1S2</i> | 0-50000 ng/mL | |
| Postdilution factor | 10 recommended b | No |
| Calc. first/last | 46/86 | 46/86 |
| Unit | ng/mL | |
| b) For use when estimating concentration in preparation for GC/MS analysis. | | |
| | | |
| Pipetting parameters | | |
| <i>AM3S2, AM3Q2</i> | | Diluent (H2O) |
| R1 | 60 µL | 20 µL |
| Sample | 10 µL | 5 µL |
| SR | 47 µL | 3 µL |
| Total volume | 145 µL | |
| | | Diluent (H2O) |
| <i>AM5S2, AM5Q2, AM5QC</i> | | |
| R1 | 60 µL | 20 µL |
| Sample | 7.5 µL | 5 µL |
| SR | 47 µL | 3 µL |
| Total volume | 142.5 µL | |
| <i>AM1S2, AM1Q2</i> | | Diluent (H2O) |
| R1 | 60 µL | 20 µL |
| Sample | 5 µL | 5 µL |
| SR | 47 µL | 3 µL |
| Total volume | 140 µL | |
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| Calibration | |
|-------------------------------|----------------------------------------------------------------------------------------------------------------------------------------|
| Calibrators | Semiquantitative applications |
| AM3S2, 0-358 | Preciset DAT Plus II calibrators |
| | 6 Calibrators (300 cutoff, DATS8, 07 6796 °) |
| | 0, 150, 300, 600, 1000, 2000 ng/mL |
| | d-methamphetamine |
| AM5S2, 0-359;<br>AM1S2, 0-360 | Preciset DAT Plus I calibrators |
| | 6 Calibrators (500 and 1000 cutoffs, DATS2, 07 6764 6 °) |
| | 0, 250, 500, 1000, 3000, 5000 ng/mL |
| | d-methamphetamine |
| | Qualitative applications |
| AM3Q2, 0-330 | 0 ng/mL (Preciset DAT Plus II calibrators, CAL 1) or deionized<br>water and 300 ng/mL (Preciset DAT Plus II calibrators, CAL 3) |
| | 2 Calibrators (300 cutoff, DATQ3, 07 6770 0°) |
| | For qualitative applications, the cutoff value is assigned as 1000 |
| AM5Q2, 0-340 | 0 ng/mL (Preciset DAT Plus I calibrators, CAL 1) or deionized<br>water and 500 ng/mL (C.f.a.s. DAT Qualitative Plus) |
| | 2 Calibrators (500 cutoff, DATQ1, 07 6744 1°) |
| | For qualitative applications, the cutoff value is assigned as 1000 |
| AM5QC, 0-341 | 0 ng/mL (Preciset DAT Plus I or II calibrators, CAL 1) or<br>deionized water and 500 ng/mL (C.f.a.s. DAT Qualitative Plus<br>Clinical) |
| | 2 Calibrators (500 cutoff, DATQ5, 07 6880 4°) |
| | For qualitative applications, the cutoff value is assigned as 1000 |
| AM1Q2, 0-350 | 0 ng/mL (Preciset DAT Plus I calibrators, CAL 1) or deionized<br>water and 1000 ng/mL (Preciset DAT Plus I calibrators, CAL 4) |
| | 2 Calibrators (1000 cutoff, DATQ2, 07 6768 9°) |
c) Short name and ID used on the COBAS INTEGRA systems.
| Calibration mode | Semiquantitative applications |
|-----------------------------|----------------------------------------------------------------------------------|
| COBAS INTEGRA 800 analyzers | Logit/log 4 |
| | Qualitative applications |
| | Linear regression |
| Calibration replicate | Duplicate recommended |
| Calibration interval | |
| COBAS INTEGRA 800 analyzers | Each lot, every 4 weeks, and as required following quality<br>control procedures |
A calibration curve is generated using the calibrators. Calibrators must be placed from the highest concentration first to the lowest last on the CAL/QC rack. This curve is retained in memory by the COBAS INTEGRA systems and recalled for later use.
· Traceability: This method has been standardized against GC/MS.
#### Quality control
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Quality control
300 ng/mL cutoff AM3S2 and AM3Q2: Control Set DAT II PreciPos DAT Set II (DAT2P, 07 6771 9 ") PreciNeg DAT Set II (DAT2N, 07 6772 7 9)
500 ng/mL cutoff AM5S2 and AM5Q2: Control Set DAT I PreciPos DAT Set I (DATIP, 07 6753 0 ª) PreciNeg DAT Set I (DATIN, 07 6754 9 d)
#### AM5QC:
Control Set DAT Clinical PreciPos DAT Clinical (DATCP, 07 6879 0 °) PreciNeg DAT Clinical (DATCN. 07 6878 2 ")
1000 ng/mL cutoff AMIS2 and AMIQ2: Control Set DAT III PreciPos DAT Set III (DAT3P, 07 6773 5 ") PreciNeg DAT Set III (DAT3N, 07 6774 3 ª) User defined Recommended
Control sequence Control after calibration
d) Short name and ID used on the COBAS INTEGRA systems.
For quality control, use control materials as listed in the Order information section. Other suitable control material can be used in addition.
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 limits. Follow the applicable government regulations and local guidelines for quality control.
#### Results
COBAS INTEGRA systems report results with the following test flags: Semiquantitative result reporting AM3S2 (300 ng/mL cutoff)
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| Flag | COBAS INTEGRA | Value range |
|-----------|---------------|--------------|
| No flag | Negative | < 300 ng/mL |
| <TEST RNG | Negative | < 0 ng/mL |
| >TEST RNG | Positive | > 2000 ng/mL |
| POS 300 | Positive | ≥ 300 ng/mL |
Value ranges listed above are based on a cutoff value of 300 ng/mL.
#### Semiquantitative result reporting
AM5S2 (500 ng/mL cutoff)
| Flag | COBAS INTEGRA | Value range |
|-----------|---------------|--------------|
| No flag | Negative | < 500 ng/mL |
| <TEST RNG | Negative | < 0 ng/mL |
| >TEST RNG | Positive | > 5000 ng/mL |
| POS 500 | Positive | ≥ 500 ng/mL |
Value ranges listed above are based on a cutoff value of 500 ng/mL.
### Semiquantitative result reporting
#### AM1S2 (1000 ng/mL cutoff)
| Flag | COBAS INTEGRA | Value range |
|-----------|---------------|--------------|
| No flag | Negative | < 1000 ng/mL |
| <TEST RNG | Negative | < 0 ng/mL |
| >TEST RNG | Positive | > 5000 ng/mL |
| POS 1000 | Positive | ≥ 1000 ng/mL |
Value ranges listed above are based on a cutoff value of 1000 ng/mL.
#### Qualitative result reporting
AM3Q2 (300 ng/mL cutoff)
| Flag | COBAS INTEGRA | Value range |
|-----------|---------------|-------------|
| No flag | Negative | < 1000 |
| <TEST RNG | Negative | < 0 |
| >TEST RNG | Positive | > 2000 |
| POS 1000 | Positive | ≥ 1000 |
Value ranges above are based on assigning the cutoff of 300 ng/mL a value of 1000.
#### Qualitative result reporting
| AM5Q2, AM5QC (500 ng/mL cutoff) | | |
|---------------------------------|---------------|-------------|
| Flag | COBAS INTEGRA | Value range |
| No flag | Negative | < 1000 |
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| <test rng<br="">дарыптананының таптының атаптынанының таптанының қалыптынын аталанын атаптанының тұрғанаттан таптынан атаптын атаптын атаптын атаптын аттыр</test> | Negative | < ( |
|--------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------|--------|
| >TEST RNG | Positive | > 2000 |
| POS 1000<br>ан жаңған жұмысындарының тұрғынан айтынша айынша айынша айынша атынын аталымының айтынша б | Positive | ≥ 1000 |
Value ranges above are based on assigning the cutoff of 500 ng/mL a value of 1000.
#### Qualitative result reporti
| AM1Q2 (1000 ng/mL cutoff) | | |
|---------------------------|---------------|-------------|
| Flag | COBAS INTEGRA | Value range |
| No flag | Negative | < 1000 |
| <TEST RNG | Negative | < 0 |
| >TEST RNG | Positive | > 2000 |
| POS 1000 | Positive | $\ge$ 1000 |
Value ranges above are based on assigning the cutoff of 1000 ng/mL a value of 1000.
#### Semiquantitative result reporting
The semiquantitation of preliminary positive results should only be used by laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GC/MS. It also permits the laboratory to establish quality control procedures and assess control performance.
Note: When using the post-dilution function (1:10 dilution), to ensure the sample was not overdiluted, the diluted result must be at least half the analyte cutoff value times 10. If the diluted result falls below half the analyte cutoff value times 10, repeat the sample with a smaller dilution. A dilution that produces a result closest to the analyte cutoff is the most accurate estimation. To estimate the positive sample's concentration, multiply the result by the appropriate dilution factor. Dilutions should only be used as an estimation for GC/MS.
#### Limitations10
See the Analytical specificity section of this method sheet for information on substances tested for cross-reactivity in 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 amphetamine or methamphetamine in urine. It does not measure the level of intoxication.
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#### ACTION REQUIRED
Special wash programming: The use of special wash steps is mandatory when certain test combinations are run together on COBAS INTEGRA 800 analyzer. Refer to the Method Manual, Introduction, Extra Wash Cycles for further instructions.
Where required, special wash/carry-over evasion programming must be implemented prior to reporting results with this test.
#### Expected values
No drug should be present in individuals that have not ingested amphetamines.
#### Specific performance data
Representative performance data on the COBAS INTEGRA 800 analyzer are given below. Results obtained in individual laboratories may differ.
#### Precision
A d-methamphetamine (MAMP) solution (1 mg/mL) was added to 9 samples obtained from a human urine 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 in qualitative and semiquantitative modes. Following a CLSI (EPS-A2) precision protocol, samples were tested in 2 replicates per run, 2 runs per day for 21 days, total N = 84.
| Qualitative - 300 ng/mL Cutoff | | | |
|--------------------------------|----------------------------|-----------------------------|--------------------------|
| Drug | Concentration of<br>Sample | Number of<br>Determinations | Results<br># Neg / # Pos |
| MAMP | zero drug | 84 | 84 Neg / 0 Pos |
| MAMP | - 75 % | 84 | 84 Neg / 0 Pos |
| MAMP | - 50 % | 84 | 84 Neg / 0 Pos |
| MAMP | - 25 % | 84 | 83 Neg / 1 Pos |
| MAMP | cutoff | 84 | 27 Neg / 57 Pos |
| MAMP | + 25 % | 84 | 0 Neg / 84 Pos |
| MAMP | + 50 % | 84 | 0 Neg / 84 Pos |
| MAMP | + 75 % | 84 | 0 Neg / 84 Pos |
| MAMP | + 100 % | 84 | 0 Neg / 84 Pos |
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| Drug | Concentration of<br>Sample | Number of<br>Determinations | Results<br># Neg / # Pos |
|------|----------------------------|-----------------------------|--------------------------|
| MAMP | zero drug | 84 | 84 Neg / 0 Pos |
| MAMP | - 75 % | 84 | 84 Neg / 0 Pos |
| MAMP | - 50 % | 84 | 84 Neg / 0 Pos |
| MAMP | - 25 % | 84 | 84 Neg / 0 Pos |
| MAMP | cutoff | 84 | 28 Neg / 56 Pos |
| MAMP | + 25 % | 84 | 0 Neg / 84 Pos |
| MAMP | + 50 % | 84 | 0 Neg / 84 Pos |
| MAMP | + 75 % | 84 | 0 Neg / 84 Pos |
| MAMP | + 100 % | 84 | 0 Neg / 84 Pos |
#### Qualitative - 500 ng/mL Cutoff
·
| Qualitative - 1000 ng/mL Cutoff | | | | | | |
|---------------------------------|----------------------------|-----------------------------|--------------------------|--|--|--|
| Drug | Concentration of<br>Sample | Number of<br>Determinations | Results<br># Neg / # Pos | | | |
| MAMP | zero drug | 84 | 84 Neg / 0 Pos | | | |
| MAMP | - 75 % | 84 | 84 Neg / 0 Pos | | | |
| MAMP | - 50 % | 84 | 84 Neg / 0 Pos | | | |
| MAMP | - 25 % | 84 | 84 Neg / 0 Pos | | | |
| MAMP | cutoff | 84 | 13 Neg / 71 Pos | | | |
| MAMP | + 25 % | 84 | 0 Neg / 84 Pos | | | |
| MAMP | + 50 % | 84 | 0 Neg / 84 Pos | | | |
| MAMP | + 75 % | 84 | 0 Neg / 84 Pos | | | |
| MAMP | + 100 % | 84 | 0 Neg / 84 Pos | | | |
{15}------------------------------------------------
| Drug | Sample<br>Conc. | Results<br># Neg /<br># Pos | Repeatability* | | Intermediate<br>Precision** | |
|------|-----------------|-----------------------------|----------------|----------|-----------------------------|----------|
| | | | SD, ng/mL | CV,<br>% | SD, ng/mL | CV,<br>% |
| MAMP | zero drug | 84 / 0 | 9.4 | 95.4 | 12.2 | 123.8 |
| MAMP | - 75 % | 84 / 0 | 12.9 | 15.5 | 15.7 | 18.8 |
| MAMP | - 50 % | 84 / 0 | 14.3 | 9.0 | 17.7 | 11.2 |
| MAMP | - 25 % | 84 / 0 | 13.2 | 5.4 | 19.8 | 8.2 |
| MAMP | cutoff | 34 / 50 | 23.2 | 7.6 | 23.5 | 7.7 |
| MAMP | + 25 % | 0 / 84 | 20.9 | 5.2 | 24.4 | 6.1 |
| MAMP | + 50 % | 0 / 84 | 22.5 | 5.0 | 29.5 | 6.6 |
| MAMP | + 75 % | 0 / 84 | 27.4 | 5.0 | 36.5 | 6.6 |
| MAMP | + 100 % | 0 / 84 | 35.7 | 5.7 | 40.8 | 6.5 |
### Semiquantitative - 300 ng/mL Cutoff
# Semiquantitative - 500 ng/mL Cutoff
| Drug | Sample<br>Conc. | Results<br># Neg /<br># Pos | Repeatability* | | Intermediate<br>Precision** | |
|------|-----------------|-----------------------------|----------------|----------|-----------------------------|----------|
| | | | SD, ng/mL | CV,<br>% | SD, ng/mL | CV,<br>% |
| MAMP | zero drug | 84 / 0 | 9.6 | 117.3 | 11.0 | 134.1 |
| MAMP | - 75 % | 84 / 0 | 14.6 | 10.5 | 20.6 | 14.9 |
| MAMP | - 50 % | 84 / 0 | 20.0 | 7.9 | 21.9 | 8.6 |
| MAMP | - 25 % | 84 / 0 | 24.1 | 6.2 | 27.2 | 7.0 |
| MAMP | cutoff | 39 / 45 | 28.2 | 5.6 | 39.4 | 7.8 |
| MAMP | + 25 % | 0 / 84 | 31.5 | 4.9 | 48.5 | 7.5 |
| MAMP | + 50 % | 0 / 84 | 34.5 | 4.8 | 49.5 | 6.8 |
| MAMP | + 75 % | 0 / 84 | 41.6 | 5.0 | 62.3 | 7.5 |
| MAMP | + 100 % | 0 / 84 | 46.7 | 4.6 | 72.4 | 7.1 |
{16}------------------------------------------------
| Drug | Sample<br>Conc. | Results<br># Neg /<br># Pos | Repeatability* | | Intermediate<br>Precision** | |
|------|-----------------|-----------------------------|----------------|----------|-----------------------------|----------|
| | | | SD, ng/mL | CV,<br>% | SD, ng/mL | CV,<br>% |
| MAMP | zero drug | 84 / 0 | 20.2 | 97.2 | 24.9 | 119.5 |
| MAMP | - 75 % | 84 / 0 | 25.3 | 9.3 | 30.7 | 11.2 |
| MAMP | - 50 % | 84 / 0 | 33.3 | 6.4 | 39.4 | 7.5 |
| MAMP | - 25 % | 84 / 0 | 34.3 | 4.6 | 52.1 | 7.0 |
| MAMP | cutoff | 25 / 59 | 56.9 | 5.5 | 71.3 | 6.9 |
| MAMP | + 25 % | 0 / 84 | 56.6 | 4.2 | 79.3 | 5.9 |
| MAMP | + 50 % | 0 / 84 | 63.6 | 4.0 | 81.3 | 5.2 |
| MAMP | + 75 % | 0 / 84 | 60.3 | 3.5 | 101.6 | 5.9 |
| MAMP | + 100 % | 0 / 84 | 108.1 | 5.1 | 126.6 | 6.0 |
#### Semiquantitative - 1000 ng/mL Cutoff
*repeatability = within-run precision
**intermediate precision = total precision / between run precision / between day precision
The same precision experiment was repeated utilizing d-amphetamine (AMP) as the target analyte instead of d-methamphetamine. The following tables show the results obtained on a COBAS INTEGRA 800 analyzer.
| Drug | Concentration of<br>Sample | Number of<br>Determinations | Results<br># Neg / # Pos |
|------|----------------------------|-----------------------------|--------------------------|
| AMP | zero drug | 84 | 84 Neg / 0 Pos |
| AMP | - 75 % | 84 | 84 Neg / 0 Pos |
| AMP | - 50 % | 84 | 84 Neg / 0 Pos |
| AMP | - 25 % | 84 | 83 Neg / 1 Pos |
| AMP | cutoff | 84 | 2 Neg / 82 Pos |
| AMP | + 25 % | 84 | 0 Neg / 84 Pos |
| AMP | + 50 % | 84 | 0 Neg / 84 Pos |
| AMP | + 75 % | 84 | 0 Neg / 84 Pos |
| AMP | + 100 % | 84 | 0 Neg / 84 Pos |
# Qualitative - 300 ng/mL Cutoff
{17}------------------------------------------------
### Qualitative - 500 ng/mL Cutoff
| Qualitative - 500 ng/mL Cutoff | | | |
|--------------------------------|----------------------------|-----------------------------|--------------------------|
| Drug | Concentration of<br>Sample | Number of<br>Determinations | Results<br># Neg / # Pos |
| AMP | zero drug | 84 | 84 Neg / 0 Pos |
| AMP | - 75 % | 84 | 84 Neg / 0 Pos |
| AMP | - 50 % | 84 | 84 Neg / 0 Pos |
| AMP | - 25 % | 84 | 82 Neg / 2 Pos |
| AMP | cutoff | 84 | 6 Neg / 78 Pos |
| AMP | + 25 % | 84 | 0 Neg / 84 Pos |
| AMP | + 50 % | 84 | 0 Neg / 84 Pos |
| AMP | + 75 % | 84 | 0 Neg / 84 Pos |
| AMP | + 100 % | 84 | 0 Neg / 84 Pos |
:
.
| Drug | Concentration of<br>Sample | Number of<br>Determinations | Results<br># Neg / # Pos |
|------|----------------------------|-----------------------------|--------------------------|
| AMP | zero drug | 84 | 84 Neg / 0 Pos |
| AMP | - 75 % | 84 | 84 Neg / 0 Pos |
| AMP | - 50 % | 84 | 84 Neg / 0 Pos |
| AMP | - 25 % | 84 | 82 Neg / 2 Pos |
| AMP | cutoff | 84 | 7 Neg / 77 Pos |
| AMP | + 25 % | 84 | 1 Neg / 83 Pos |
| AMP | + 50 % | 84 | 0 Neg / 84 Pos |
| AMP | + 75 % | 84 | 0 Neg / 84 Pos |
| AMP | + 100 % | 84 | 0 Neg / 84 Pos |
.
・
.
{18}------------------------------------------------
| Drug | Sample<br>Conc. | Results<br># Neg /<br># Pos | Repeatability* | | Intermediate<br>Precision** | |
|------|-----------------|-----------------------------|----------------|----------|------------------------------|----------|
| | | | SD, ng/mL | CV,<br>% | SD, ng/mL | CV,<br>% |
| AMP | zero drug | 84 / 0 | 8.6 | 110.0 | 10.6 | 135.7 |
| AMP | - 75 % | 84 / 0 | 10.4 | 16.8 | 13.6 | 21.9 |
| AMP | - 50 % | 84 / 0 | 14.6 | 10.0 | 19.9 | 13.6 |
| AMP | - 25 % | 84 / 0 | 15.7 | 6.6 | 20.6 | 8.7 |
| AMP | cutoff | 4 / 80 | 25.8 | 7.4 | 28.5 | 8.2 |
| AMP | + 25 % | 0 / 84 | 30.7 | 7.1 | 32.2 | 7.4 |
| AMP | + 50 % | 0 / 84 | 31.1 | 6.1 | 37.6 | 7.3 |
| AMP | + 75 % | 0 / 84 | 29.1 | 4.9 | 41.2 | 7.0 |
| AMP | + 100 % | 0 / 84 | 36.2 | 5.3 | 44.6 | 6.5 |
| Drug | Sample<br>Conc. | Results<br># Neg<br># Pos | Repeatability* | | Intermediate<br>Precision ** | |
| | | | SD, ng/mL | CV,<br>% | SD, ng/mL | CV,<br>% |
| AMP | zero drug | 84 / 0 | 12.6 | 117.0 | 13.2 | 122.5 |
| AMP | - 75 % | 84 / 0 | 21.4 | 16.9 | 25.2 | 20.0 |
| AMP | - 50 % | 84 / 0 | 18.2 | 6.9 | 25.6 | 9.8 |
| AMP | - 25 % | 83 / 1 | 25.6 | 5.7 | 29.7 | 6.6 |
| AMP | cutoff | 0 / 84 | 29.1 | 4.7 | 38.3 | 6.2 |
| AMP | + 25 % | 0-/ 84 | 42.0 | ર. ર | 51.7 | 6.8 |
| AMP | + 50 % | 0 / 84 | 40.3 | 4.6 | 49.9 | 5.7 |
| AMP | + 75 % | 0 / 84 | 58.7 | 5.9 | 59.7 | 6.0 |
| AMP | + 100 % | 0 / 84 | 79.0 | 7.0 | 86.3 | 7 6 |
### Semiquantitative - 300 ng/mL Cutoff
100 - 100 - 100 -
:
:
·
:
.
{19}------------------------------------------------
| Drug | Sample<br>Conc. | Results<br># Neg /<br># Pos | Repeatability* | | Intermediate<br>Precision ** | |
|------|-----------------|-----------------------------|----------------|-----------|------------------------------|------------|
| | | | SD, ng/mL | CV,<br>0% | SD, ng/mL | CV.<br>0/0 |
| AMP | zero drug | 84 / 0 | ાં 5.9 | 】】【 l | 19.0 | 132.6 |
| AMP | - 75 % | 84 / 0 | 21.9 | 9.2 | 29.8 | 12.5 |
| AMP | - 50 % | 84 / 0 | 30.3 | 5.4 | 40.9 | 7.3 |
| AMP | - 25 % | 84 / 0 | 44.3 | 5.2 | રરે :0 | 6.4 |
| AMP | cutoff | 6 / 78 | રૂડે તે | 4.8 | 84.3 | 7.6 |
| AMP | + 25 % | 0 / 84 | 72.5 | ર્ટ ર્ | 87 ર | 6.7 |
| AMP | + ૨૦ % | 0 / 84 | 65.7 | 4.4 | તેરું ર | ર્ણ રે |
| AMP | + 75 % | 0 / 84 | 74.3 | 4.5 | 105.2 | 64 |
| AMP | + 100 % | 0 / 84 | 106.7 | ર તેવે | 131.4 | 7.3 |
#### Semiquantitative - 1000 no/mL Cutoff
*repeatability = within-run precision
** intermediate precision = total precision / between run precision / between day precision
#### Accuracy
The accuracy of this assay was determined against d-methamphetamine or d-amphetamine GC/MS results. The evaluated cutoff concentrations for the GC/MS testing were the same as the assay screening cutoffs. For both d-methamphetamine, 36 urine samples, obtained from a clinical laboratory were confirmed to be negative by GC/MS and were evaluated with Amphetamines II. 100 % of these normal urines were negative with both the semiguantitative and qualitative assay relative to the 300 ng/mL, and 1000 ng/mL cutoffs. 4 additional unaltered samples, containing d-methamphetamine between 85-204 ng/mL (GC/MS confirmed), were tested with the 500 ng/mL cutoff and gave negative results.
36 samples obtained from a clinical laboratory, where they screened positive with a commercially available enzyme immunoassay and were subsequently confirmed positive by GC/MS to contain d-methamphetamine or d-amphetamine, were evaluated with Amphetamines II. 100% of these samples were positive relative to the 300 ng/mL, and 1000 ng/mL, cutoffs.
In addition, several unaltered near cutoff samples were run for each cutoff. These samples fell in the near cutoff negative range (between - 50 % and cutoff) and the near cutoff positive range (between cutoff and + 50 %). For each cutoff, 4 negative near cutoff samples and 4 positive near cutoff samples were assayed.
Data from the accuracy studies described above were combined with data generated from the unaltered near cutoff urine samples. The following results were obtained with the Amphetamines II assay on the COBAS INTEGRA 800 analyzer relative to the GC/MS values for both d-methamphetamine and d-amphetamine.
{20}------------------------------------------------
## Amphetamines II Qualitative Assay Results (MAMP)
| Roche<br>ONLINE<br>DAT AMPII<br>assay | Low Neg | Near Cutoff<br>Negative by<br>GC/MS<br>(between -<br>50 % and<br>cutoff) | Near Cutoff<br>Positive by<br>GC/MS<br>(between cutoff<br>and + 50 %) | High Positive<br>by GC/MS<br>(greater than<br>+ 50 %) | Percent<br>Agreement with<br>GC/MS (MAMP) |
|---------------------------------------|---------|--------------------------------------------------------------------------|-----------------------------------------------------------------------|-------------------------------------------------------|-------------------------------------------|
| 300 ng/mL Cutoff | | | | | |
| Positive | 0 | 4 | 4 | 36 | 100 % |
| Negative | 36 | 0 | 0 | 0 | 90 % |
| 500 ng/mL Cutoff | | | | | |
| Positive | 0 | 4 | 4 | 36 | 100 % |
| Negative | 40 | 0 | 0 | 0 | 91 % |
| 1000 ng/mL Cutoff | | | | | |
| Positive | 0 | 4 | 4 | 36 | 100 % |
| Negative | 36 | 0 | 0 | 0 | 90 % |
# Amphetamines II Semiquantitative Assay Results (MAMP)
| Roche<br>ONLINE<br>DAT AMPΙΙ<br>assay | Low Neg | Near Cutoff<br>Negative by<br>GC/MS<br>(between -<br>50 % and<br>cutoff) | Near Cutoff<br>Positive by<br>GC/MS<br>(between cutoff<br>and + 50 %) | High Positive<br>by GC/MS<br>(greater than<br>+ 50 %) | Percent<br>Agreement with<br>GC/MS (MAMP) |
|---------------------------------------|---------|--------------------------------------------------------------------------|-----------------------------------------------------------------------|-------------------------------------------------------|-------------------------------------------|
| 300 ng/mL Cutoff | | | | | |
| Positive | 0 | 4 | 4 | 36 | 100 % |
| Negative | 36 | 0 | 0 | 0 | 90 % |
| 500 ng/mL Cutoff | | | | | |
| Positive | 0 | 4 | 4 | 36 | 100 % |
| Negative | 40 | 0 | 0 | 0 | 91 % |
| 1000 ng/mL Cutoff | | | | | |
| Positive | 0 | 4 | 4 | 36 | 100 % |
| Negative | 36 | 0 | 0 | 0 | 90 % |
.
{21}------------------------------------------------
Accuracy samples were categorized based upon the d-methamphetamine GC/MS concentration only. The table below identifies those samples with a d-methamphetamine concentration below the cutoff, in which the observed result on a COBAS INTEGRA 800 analyzer was positive. The expected results column identifies the result expected with the Amphetamines II assay based upon the d-methamphetamine (MAMP) value relative to the cutoff.
| Cutoff Value<br>(ng/mL) | Roche ONLINE<br>DAT AMP II<br>OBSERVED<br>Result | Roche ONLINE<br>DAT AMP II<br>EXPECTED<br>Result | GC/MS<br>(ng/mL) | Drug / Metabolite |
|-------------------------|--------------------------------------------------|--------------------------------------------------|------------------|-------------------|
| 300 (SQ, Q) | Positive | Positive | 173<br>181 | MAMP<br>AMP |
| 300 (SQ, Q) | Positive | Positive | 278<br>101 | MAMP<br>AMP |
| 300 (SQ, Q) | Positive | Positive | 171<br>220 | MAMP<br>AMP |
| 300 (SQ, Q) | Positive | Positive | 291<br>145 | MAMP<br>AMP |
| 500 (SQ, Q) | Positive | Positive | 488<br>466 | MAMP<br>AMP |
| 500 (SQ, Q) | Positive | Negative | 325<br>171 | MAMP<br>AMP |
| 500 (SQ, Q) | Positive | Negative | 291<br>145 | MAMP<br>AMP |
| 500 (SQ, Q) | Positive | Positive | 472<br>650 | MAMP<br>AMP |
| 1000 (SQ, Q) | Positive | Positive | 706<br>443 | MAMP<br>AMP |
| 1000 (SQ, Q) | Positive | Positive | 540<br>693 | MAMP<br>AMP |
| 1000 (SQ, Q) | Positive | Positive | 769<br>395 | MAMP<br>AMP |
| 1000 (SQ, Q) | Positive | Positive | 572<br>432 | MAMP<br>AMP |
# GC/MS Summary of Discrepant Results (MAMP)
{22}------------------------------------------------
## Amphetamines II Qualitative Assay Results (AMP)
·
:
. 1
| Roche<br>ONLINE<br>DAT AMPII<br>assay | Low Neg | Near Cutoff<br>Negative by<br>GC/MS<br>(between -<br>50 % and<br>cutoff) | Near Cutoff<br>Positive by<br>GC/MS<br>(between cutoff<br>and + 50 %) | High Positive<br>by GC/MS<br>(greater than<br>+ 50 %) | Percent<br>Agreement with<br>GC/MS (AMP) |
|---------------------------------------|---------|--------------------------------------------------------------------------|-----------------------------------------------------------------------|-------------------------------------------------------|------------------------------------------|
| 300 ng/mL Cutoff | | | | | |
| Positive | 0 | 3 | 4 | 36 | 100 % |
| Negative | 36 | 1 | 0 | 0 | 92.5 % |
| 500 ng/mL Cutoff | | | | | |
| Positive | 0 | 3 | 4 | 36 | 100 % |
| Negative | 36 | 1 | 0 | 0 | 92.5 % |
| 1000 ng/mL Cutoff | | | | | |
| Positive | 0 | 1 | 4 | 36 | 100 % |
| Negative | 36 | 3 | 0 | 0 | 97.5 % |
. .
# Amphetamines II Semiquantitative Assay Results (AMP)
:
. .
| Roche<br>ONLINE<br>DAT AMPII<br>assay | Low Neg | Near Cutoff<br>Negative by<br>GC/MS<br>(between -<br>50 % and<br>cutoff) | Near Cutoff<br>Positive by<br>GC/MS<br>(between cutoff<br>and + 50 %) | High Positive<br>by GC/MS<br>(greater than<br>+ 50 %) | Percent<br>Agreement with<br>GC/MS (AMP) |
|---------------------------------------|---------|--------------------------------------------------------------------------|-----------------------------------------------------------------------|-------------------------------------------------------|------------------------------------------|
| 300 ng/mL Cutoff | | | | | |
| Positive | 0 | 3 | 4 | 36 | 100 % |
| Negative | 36 | 1 | 0 | 0 | 92.5 % |
| 500 ng/mL Cutoff | | | | | |
| Positive | 0 | 3 | 4 | 36 | 100 % |
| Negative | 36 | 1 | 0 | 0 | 92.5 % |
| 1000 ng/mL Cutoff | | | | | |
| Positive | 0 | 2 | 4 | 36 | 100 % |
| Negative | 36 | 2 | 0 | 0 | 95 % |
{23}------------------------------------------------
Accuracy samples were categorized based upon the d-amphetamine GC/MS concentration only. The table below identifies those samples with a d-amphetamine concentration below the cutoff, in which the observed result on a COBAS INTEGRA 800 analyzer was positive. The expected results column identifies the result expected with the Amphetamines II assay based upon the damphetamine (AMP) value relative to the cutoff.
| Cutoff Value<br>(ng/mL) | Roche ONLINE<br>DAT AMP II<br>OBSERVED<br>Result | Roche ONLINE<br>DAT AMP II<br>EXPECTED<br>Result | GC/MS<br>(ng/mL) | Drug / Metabolite |
|-------------------------|--------------------------------------------------|--------------------------------------------------|------------------|-------------------|
| 300 (SQ, Q) | Positive | Positive | 157<br>363 | AMP<br>MAMP |
| 300 (SQ, Q) | Positive | Positive | 181<br>173 | AMP<br>MAMP |
| 300 (SQ, Q) | Positive | Positive | 220<br>171 | AMP<br>MAMP |
| 500 (SQ, Q) | Positive | Positive | 438<br>121 | AMP<br>MAMP |
| 500 (SQ, Q) | Positive | Positive | 457<br>1152 | AMP<br>MAMP |
| 500 (SQ, Q) | Positive | Positive | 443<br>706 | AMP<br>MAMP |
| 1000 (SQ) | Positive | Negative | 920 | AMP |
| 1000 (SQ, Q) | Positive | Positive | 837<br>1163 | AMP<br>MAMP |
### GC/MS Summary of Discrepant Results (AMP)
#### Analytical specificity
The specificity of Amphetamines II, on the COBAS INTEGRA 800 analyzer, for various phenethylamines and structurally similar compounds was determined by generating inhibition curves for each of the compounds listed for both semiquantitative modes and determining the approximate quantity of each compound that is equivalent in assay reactivity to the 300 ng/mL, 500 ng/mL, and 1000 ng/mL d-methamphetamine assay cutoff. The tables below show the semiquantitative results of the study for each assay cutoff. The same samples were run in the qualitative mode and all recovered appropriately negative or positive, based on the calculated cross-reactivity.
{24}------------------------------------------------
| Compound | ng/mL<br>Equivalent to<br>300 ng/mL<br>MAMP | Approx. Percent<br>Cross-reactivity |
|-------------------------------------|---------------------------------------------|-------------------------------------|
| ± MDMAc | 114 | 264 |
| ± MDAf | 249 | 121 |
| ± MDEAh | 285 | 105 |
| <i>d</i> -Amphetamine | 311 | 96 |
| <i>d</i> -Methamphetamine | 327 | 92 |
| ± MBDB HClg | 339 | 88 |
| ± BDB HCli | 648 | 46 |
| <i>l</i> -Methamphetamine | 2754 | 11 |
| <i>l</i> -Amphetamine | 6443 | 5 |
| Phendimetrazine | 47473 | 0.63 |
| Phentermine | 66385 | 0.45 |
| Tyramine | 86271 | 0.35 |
| <i>d</i> -Pseudoephedrine | 87578 | 0.34 |
| <i>l</i> -Ephedrine | 94792 | 0.32 |
| <i>d,l</i> -Phenylpropanolamine HCl | 280374 | 0.11 |
| <i>d</i> -Ephedrine | 329670 | 0.09 |
:
and the country of the county of the county of
{25}------------------------------------------------
| Compound | ng/mL<br>Equivalent to<br>500 ng/mL<br>MAMP | Approx. Percent<br>Cross-reactivity |
|-------------------------------------|---------------------------------------------|-------------------------------------|
| ± MDMAc | 173 | 289 |
| ± MDAf | 433 | 115 |
| <i>d</i> -Methamphetamine | 444 | 113 |
| <i>d</i> -Amphetamine | 460 | 109 |
| ± MDEAh | 494 | 101 |
| ± MBDB HClg | 713 | 70 |
| ± BDB HCli | 1209 | 41 |
| <i>l</i> -Methamphetamine | 4098 | 12 |
| <i>l</i> -Amphetamine | 11174 | 4 |
| Phendimetrazine | 72500 | 0.69 |
| Phentermine | 118483 | 0.42 |
| <i>d</i> -Pseudoephedrine | 132275 | 0.38 |
| Tyramine | 141243 | 0.35 |
| <i>l</i> -Ephedrine | 154321 | 0.32 |
| <i>d,l</i> -Phenylpropanolamine HCl | 390625 | 0.13 |
| <i>d</i> -Ephedrine | 413223 | 0.12 |
.
and the comments of the comments of the comments of the comments of
.
{26}------------------------------------------------
| Compound | ng/mL<br>Equivalent to<br>1000 ng/mL<br>MAMP | Approx. Percent<br>Cross-reactivity |
|--------------------------------------|----------------------------------------------|-------------------------------------|
| ± MDMAc | 446 | 224 |
| ± MDAf | 785 | 127 |
| <i>d</i> -Methamphetamine | 970 | 103 |
| <i>d</i> -Amphetamine | 1024 | 98 |
| ± MBDB HClg | 1194 | 84 |
| ± MDEAh | 1203 | 83 |
| ± BDB HCli | 2262 | 44 |
| <i>l</i> -Methamphetamine | 9008 | 11 |
| <i>l</i> -Amphetamine | 23445 | 4 |
| Phendimetrazine | 156740 | 0.64 |
| <i>d</…
