Predicate Devices

Reference Devices

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description details a standard immunoassay based on chemical reactions and absorbance measurements, with no mention of AI or ML algorithms for data processing or interpretation.

Therapeutic

No
This device is an in vitro diagnostic test used to detect substances (amphetamines and methamphetamines) in human urine. It provides analytical results and does not directly treat any medical condition.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that "Amphetamines II (AMPII) is an in vitro diagnostic test."

SaMD (Software as a Medical Device)

No

The device description clearly states it consists of "two wet reagents; a soluble drug-conjugate, and an antibody-bound microparticle solution," which are physical components, not software. It is an in vitro diagnostic test that uses immunoassay technology.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use/Indications for Use: The document explicitly states "Amphetamines II (AMPII) is an in vitro diagnostic test for the qualitative and semiquantitative detection of amphetamines and methamphetamines in human urine". This directly aligns with the definition of an in vitro diagnostic device, which is intended for use in the diagnosis of disease or other conditions, including a determination of the state of health, in order to cure, mitigate, treat, or prevent disease or its sequelae. In this case, it's used to detect the presence of substances in a sample taken from the human body (urine).
Device Description: The description details an "immunoassay for use on automated clinical chemistry analyzers" and describes the reagents and how they interact with a human urine sample to produce a result. This is characteristic of an IVD.
Performance Studies: The document includes performance studies (Accuracy and Precision) conducted on human urine samples, which are standard for demonstrating the performance of an IVD.
Predicate Device(s): The mention of predicate devices with K numbers (K083764) further indicates that this device is being compared to other legally marketed IVDs.

Therefore, based on the provided information, the Amphetamines II device is clearly an In Vitro Diagnostic.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for 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. Gas chromatography/mass spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgement should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

Product codes

DKZ, LAF

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.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Precision:
Study type: Precision study
Sample size: 9 samples with varying concentrations, tested in 2 replicates per run, 2 runs per day for 21 days, total N = 84 for each concentration level.
Key results: Presented in tables for qualitative and semiquantitative modes at 300, 500, and 1000 ng/mL cutoffs for both d-methamphetamine (MAMP) and d-amphetamine (AMP), showing number of negative/positive results, standard deviation (SD), and coefficient of variation (CV) for repeatability and intermediate precision.

Accuracy:
Study type: Accuracy study against GC/MS results.
Sample size:

36 human urine samples confirmed negative by GC/MS.
4 additional unaltered samples containing d-methamphetamine between 85-204 ng/mL (GC/MS confirmed), tested with 500 ng/mL cutoff.
36 samples that screened positive with a commercial enzyme immunoassay and were confirmed positive by GC/MS for d-methamphetamine or d-amphetamine.
Several unaltered near cutoff samples for each cutoff (4 negative near cutoff and 4 positive near cutoff samples for each cutoff).
Key results: Percentage agreement with GC/MS is reported for qualitative and semiquantitative assays at 300, 500, and 1000 ng/mL cutoffs for both MAMP and AMP. For MAMP, percent agreement ranges from 90% to 100%. For AMP, percent agreement ranges from 92.5% to 100%. Tables also show discrepant results where the observed result was positive although the GC/MS concentration was below the cutoff.

Analytical specificity:
Study type: Cross-reactivity study.
Key results: Tables show approximate percent cross-reactivity for various phenethylamines and structurally similar compounds at 300, 500, and 1000 ng/mL MAMP equivalent cutoffs.

Interference and cross-reactivity with unrelated drugs:
Study type: Interference study.
Key results: Unrelated compounds added at 100000 ng/mL to human urine showed no interference with low and high controls for all cutoffs in both semiquantitative and qualitative modes, with the exception of LSD at 2500 ng/mL, which showed 0.32% and 0.71% cross-reactivity for 300 ng/mL and 1000 ng/mL cutoffs respectively. No interference was found for tested compounds at specific concentrations for qualitative modes. In semiquantitative mode, all negative and positive controls recovered properly (within 80% - 120%) in the presence of the interfering substance. No control cross-overs were observed for samples with specific gravities ranging from 1.001 to 1.034 or pH ranging from 4.5 to 8.0 (≤ 5% cross-overs).

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Qualitative and Semiquantitative assay results show:

At 300 ng/mL, 500 ng/mL, and 1000 ng/mL cutoffs for MAMP: 100% of Positive results agreed with GC/MS, 90% or 91% of Negative results agreed with GC/MS.
At 300 ng/mL and 500 ng/mL cutoffs for AMP: 100% of Positive results agreed with GC/MS, 92.5% of Negative results agreed with GC/MS.
At 1000 ng/mL cutoff for AMP: 100% of Positive results agreed with GC/MS, 97.5% (qualitative) and 95% (semiquantitative) of Negative results agreed with GC/MS.

Predicate Device(s)

K083764

Reference Device(s)

K060645, K080183

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 862.3100 Amphetamine test system.

(a)
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.(b)
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).

Summary

0

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

1

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.

2

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.

3

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
1. a soluble drug-polymer conjugate
1. increased sensitivity to "designer drugs"and their metabolites, and
1. 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).

4

Comparison of Technological Characteristics
Feature	Amphetamines II
Assay, Integra 800	Predicate Device: Amphetamines II
Assay, Hitachi 917 (K083764)
Methodology	Same	KIMS, Kinetic interaction of
microparticles in solution
Sample Type	Same	Urine
Intended Use	Same	Qualitative and semi-quantitative
detection of amphetamine and
methamphetamine
Reagent	Same	1. Conjugate Working Solution: Conjugated
amphetamine and methamphetamine
derivatives in buffer with bovine serum
albumin(BSA) and 0.09% sodium azide.

Antibody/Microparticle Working Solution:
Microparticles attached to amphetamine and
methamphetamine antibodies (mouse
monoclonal) in buffer with bovine serum
albumin (BSA) and 0.09% sodium azide. | | |
| Cutoff | Same | 300, 500, 1000 ng/mL | | |
| Calibrator/Control
Matrix | Same | Human urine | | |

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5

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

6

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

7

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

8

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
AM3S2	0-2000 ng/mL	0-2000
AM5S2, AM1S2	0-5000 ng/mL
with postdilution
AM3S2	0-20000 ng/mL
AM5S2, AM1S2	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
AM3S2, AM3Q2		Diluent (H2O)
R1	60 µL	20 µL
Sample	10 µL	5 µL
SR	47 µL	3 µL
Total volume	145 µL
		Diluent (H2O)
AM5S2, AM5Q2, AM5QC
R1	60 µL	20 µL
Sample	7.5 µL	5 µL
SR	47 µL	3 µL
Total volume	142.5 µL
AM1S2, AM1Q2		Diluent (H2O)
R1	60 µL	20 µL
Sample	5 µL	5 µL
SR	47 µL	3 µL
Total volume	140 µL

9

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;
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
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
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
deionized water and 500 ng/mL (C.f.a.s. DAT Qualitative Plus
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
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
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

10

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)

11

Flag	COBAS INTEGRA	Value range
No flag	Negative	TEST RNG
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	TEST RNG
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	TEST RNG
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	TEST RNG
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	дарыптананының таптының атаптынанының таптанының қалыптынын аталанын атаптанының тұрғанаттан таптынан атаптын атаптын атаптын атаптын аттыр
POS 1000
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	TEST RNG
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.

13

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
Sample	Number of
Determinations	Results

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 |

14

| Drug | Concentration of
Sample | Number of
Determinations | Results

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
Sample	Number of
Determinations	Results

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
Conc. | Results

Neg /

Pos | Repeatability* | | Intermediate

Precision** | |
|------|-----------------|-----------------------------|----------------|----------|-----------------------------|----------|
| | | | SD, ng/mL | CV,
% | SD, ng/mL | CV,
% |
| 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
Conc. | Results

Neg /

Pos | Repeatability* | | Intermediate

Precision** | |
|------|-----------------|-----------------------------|----------------|----------|-----------------------------|----------|
| | | | SD, ng/mL | CV,
% | SD, ng/mL | CV,
% |
| 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
Conc. | Results

Neg /

Pos | Repeatability* | | Intermediate

Precision** | |
|------|-----------------|-----------------------------|----------------|----------|-----------------------------|----------|
| | | | SD, ng/mL | CV,
% | SD, ng/mL | CV,
% |
| 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
Sample | Number of
Determinations | Results

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
Sample	Number of
Determinations	Results

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
Sample | Number of
Determinations | Results

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
Conc. | Results

Neg /

Pos | Repeatability* | | Intermediate

Precision** | |
|------|-----------------|-----------------------------|----------------|----------|------------------------------|----------|
| | | | SD, ng/mL | CV,
% | SD, ng/mL | CV,
% |
| 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
Conc. | Results

Neg

Pos | Repeatability* | | Intermediate

Precision ** | |
| | | | SD, ng/mL | CV,
% | SD, ng/mL | CV,
% |
| 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
Conc. | Results

Neg /

Pos | Repeatability* | | Intermediate

Precision ** | |
|------|-----------------|-----------------------------|----------------|-----------|------------------------------|------------|
| | | | SD, ng/mL | CV,
0% | SD, ng/mL | CV.
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)

50 %) | Percent
Agreement with
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)

50 %) | Percent
Agreement with
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
(ng/mL) | Roche ONLINE
DAT AMP II
OBSERVED
Result | Roche ONLINE
DAT AMP II
EXPECTED
Result | GC/MS
(ng/mL) | Drug / Metabolite |
|-------------------------|--------------------------------------------------|--------------------------------------------------|------------------|-------------------|
| 300 (SQ, Q) | Positive | Positive | 173
181 | MAMP
AMP |
| 300 (SQ, Q) | Positive | Positive | 278
101 | MAMP
AMP |
| 300 (SQ, Q) | Positive | Positive | 171
220 | MAMP
AMP |
| 300 (SQ, Q) | Positive | Positive | 291
145 | MAMP
AMP |
| 500 (SQ, Q) | Positive | Positive | 488
466 | MAMP
AMP |
| 500 (SQ, Q) | Positive | Negative | 325
171 | MAMP
AMP |
| 500 (SQ, Q) | Positive | Negative | 291
145 | MAMP
AMP |
| 500 (SQ, Q) | Positive | Positive | 472
650 | MAMP
AMP |
| 1000 (SQ, Q) | Positive | Positive | 706
443 | MAMP
AMP |
| 1000 (SQ, Q) | Positive | Positive | 540
693 | MAMP
AMP |
| 1000 (SQ, Q) | Positive | Positive | 769
395 | MAMP
AMP |
| 1000 (SQ, Q) | Positive | Positive | 572
432 | MAMP
AMP |

GC/MS Summary of Discrepant Results (MAMP)

22

Amphetamines II Qualitative Assay Results (AMP)

·

:

. 1

50 %) | Percent
Agreement with
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)

:

. .

50 %) | Percent
Agreement with
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
(ng/mL) | Roche ONLINE
DAT AMP II
OBSERVED
Result | Roche ONLINE
DAT AMP II
EXPECTED
Result | GC/MS
(ng/mL) | Drug / Metabolite |
|-------------------------|--------------------------------------------------|--------------------------------------------------|------------------|-------------------|
| 300 (SQ, Q) | Positive | Positive | 157
363 | AMP
MAMP |
| 300 (SQ, Q) | Positive | Positive | 181
173 | AMP
MAMP |
| 300 (SQ, Q) | Positive | Positive | 220
171 | AMP
MAMP |
| 500 (SQ, Q) | Positive | Positive | 438
121 | AMP
MAMP |
| 500 (SQ, Q) | Positive | Positive | 457
1152 | AMP
MAMP |
| 500 (SQ, Q) | Positive | Positive | 443
706 | AMP
MAMP |
| 1000 (SQ) | Positive | Negative | 920 | AMP |
| 1000 (SQ, Q) | Positive | Positive | 837
1163 | AMP
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
Equivalent to
300 ng/mL
MAMP | Approx. Percent
Cross-reactivity |
|-------------------------------------|---------------------------------------------|-------------------------------------|
| ± MDMAc | 114 | 264 |
| ± MDAf | 249 | 121 |
| ± MDEAh | 285 | 105 |
| d -Amphetamine | 311 | 96 |
| d -Methamphetamine | 327 | 92 |
| ± MBDB HClg | 339 | 88 |
| ± BDB HCli | 648 | 46 |
| l -Methamphetamine | 2754 | 11 |
| l -Amphetamine | 6443 | 5 |
| Phendimetrazine | 47473 | 0.63 |
| Phentermine | 66385 | 0.45 |
| Tyramine | 86271 | 0.35 |
| d -Pseudoephedrine | 87578 | 0.34 |
| l -Ephedrine | 94792 | 0.32 |
| d,l -Phenylpropanolamine HCl | 280374 | 0.11 |
| d -Ephedrine | 329670 | 0.09 |

:

and the country of the county of the county of

25

| Compound | ng/mL
Equivalent to
500 ng/mL
MAMP | Approx. Percent
Cross-reactivity |
|-------------------------------------|---------------------------------------------|-------------------------------------|
| ± MDMAc | 173 | 289 |
| ± MDAf | 433 | 115 |
| d -Methamphetamine | 444 | 113 |
| d -Amphetamine | 460 | 109 |
| ± MDEAh | 494 | 101 |
| ± MBDB HClg | 713 | 70 |
| ± BDB HCli | 1209 | 41 |
| l -Methamphetamine | 4098 | 12 |
| l -Amphetamine | 11174 | 4 |
| Phendimetrazine | 72500 | 0.69 |
| Phentermine | 118483 | 0.42 |
| d -Pseudoephedrine | 132275 | 0.38 |
| Tyramine | 141243 | 0.35 |
| l -Ephedrine | 154321 | 0.32 |
| d,l -Phenylpropanolamine HCl | 390625 | 0.13 |
| d -Ephedrine | 413223 | 0.12 |

.

and the comments of the comments of the comments of the comments of

.

26

| Compound | ng/mL
Equivalent to
1000 ng/mL
MAMP | Approx. Percent
Cross-reactivity |
|--------------------------------------|----------------------------------------------|-------------------------------------|
| ± MDMAc | 446 | 224 |
| ± MDAf | 785 | 127 |
| d -Methamphetamine | 970 | 103 |
| d -Amphetamine | 1024 | 98 |
| ± MBDB HClg | 1194 | 84 |
| ± MDEAh | 1203 | 83 |
| ± BDB HCli | 2262 | 44 |
| l -Methamphetamine | 9008 | 11 |
| l -Amphetamine | 23445 | 4 |
| Phendimetrazine | 156740 | 0.64 |
| d -Pseudoephedrine | 269542 | 0.37 |
| Phentermine | 294118 | 0.34 |
| l -Ephedrine | 317460 | 0.32 |
| Tyramine | 323625 | 0.31 |
| d -Ephedrine | 793651 | 0.13 |
| d, l -Phenylpropanolamine HCl | 1111111 | 0.09 |

e) d,l-3,4-Methylenedioxymethamphetamine

f) d,I-3,4-Methylenedioxyamphetamine

g ) d J-N-Methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine hydrochloride

h) d, l-3,4-Methylenedioxyethylamphetamine

i) d,/-3,4-Methylenedioxyphenyl-2-butanamine hydrochloride

27

Interference and cross-reactivity with unrelated drugs

The following compounds were added at the listed concentrations to a human urine pool spiked with either d-amphetamine or d-methamphetamine at approximately the negative and positive control concentrations for each cutoff (+/- 25 % of assay cutoff). For each compound, the control level samples recovered properly for the 300 ng/mL, 500 ng/mL, and 1000 ng/mL cutoff in both semiquantitative and qualitative modes.

| | All Cutoffs | | All Cutoffs | |
|-------------------------|------- | Compound | Acetaminophen | 100000 | Acetylsalicylic acid | 100000 | Amitriptyline | 100000 | Aspartame | 40000 | Benzocaine | 100000 | Benzoylecgonine | 100000 | Caffeine | 100000 | Cannabidiol | 100000 | Cocaine | 100000 | Codeine | 100000 | Desipramine HCl | 100000 | Dextromethorphan | 100000 | Dextropropoxyphene | 100000 | Diazepam | 100000 | Digoxin | 100000 | Diphenhydramine | 100000 | Diphenylhydantoin | 100000 | Doxepin | 100000 | Ecgonine | 100000 | Ecgonine methyl ester | 100000 | Erythromycin | 100000 | Furosemide | 100000 | Guaiacol glycerol ether | 100000 | Hydrochlorothiazide | 100000 | Ibuprofen | 100000 | Ketamine | 100000 | Levothyroxine | 100000 | LSD | 2500 | Meperidine | 100000 | Methadone | 100000 | Methaqualone | 75000 | Morphine | 100000 | Naloxone | 100000 | Naltrexone | 100000 | Naproxen | 100000 | Niacinamide | 100000 | Nicotine | 100000 | Nifedipine | 100000 | Nordiazepam | 100000 | Omeprazole | 100000 | Oxazepam | 100000 | Penicillin G | 100000 | Phencyclidine | 40000 | Phenobarbital | 100000 | Quinine | 100000 | Secobarbital | 100000 | Tetracycline | 100000 | Δ9-THC | 10000 | Semiquantitative
Qualitative
--------|---------------------------------|-----------|----------------------------|-----------|
| Conc. (ng/mL) | Low Ctrl | High Ctrl | Low Ctrl | High Ctrl |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |
| NEG | POS | NEG | POS |

28

The compounds were additionally added to aliquots of pooled drug-free human urine at a concentration of 100000 ng/mL. None of these compounds gave values in the assay that were equal to or greater than 0.19 % cross-reactivity and no results were greater than the assay cutoffs (300 ng/mL, 500 ng/mL, and 1000 ng/mL), with the following exception.

The cross-reactivity for LSD was tested at a concentration of 2500 ng/mL. The results obtained were 0.32 %, and 0.71 %, for the 300 ng/mL and 1000 ng/mL assay cutoffs respectively.

Interference with other substances

Interfering substances were added to urine containing d-methamphetamine (MAMP) at - 25 % and + 25 % of the cutoff level at the concentration listed below. The same substances were additionally added to urine containing d-amphetamine (AMP) at - 25 % and + 25 % of the cutoff level at the concentration listed below. All samples were tested and the following results were obtained on a COBAS INTEGRA 800 analyzer. The value in the table indicates the level at which no interference was found for samples containing either d-methamphetamine or d-amphetamine.

Qualitative		300 ng/mL Cutoff		500 ng/mL Cutoff		1000 ng/mL Cutoff
Compound	Cmpd.
Conc.	Neg
Level	Pos
Level	Neg
Level	Pos
Level	Neg
Level	Pos
Level
Acetone	7.9 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Ascorbic Acid	10 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Conjugated
Bilirubin	0.1 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Creatinine	5 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Ethanol	7.9 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Glucose	12 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Hemoglobin	1 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Human serum
albumin	3 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Oxalic Acid	2 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
Sodium Chloride	23 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos

29

Urea	60 mg/mL	Neg	Pos	Neg	Pos	Neg	Pos
------	----------	-----	-----	-----	-----	-----	-----

The same experiment was performed in the semiquantitative mode for each cutoff. All negative and positive controls recovered properly, within 80 % - 120 %, in the presence of the interfering substance.

An additional protocol was executed in which samples containing either AMP or MAMP at control levels (± 25 % of cutoff) with specific gravities ranging from 1.001 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.

Samples having a pH ranging from 4.5 to 8.0 and containing either AMP or MAMP at control levels (+ 25 % of cutoff) were also tested. There were ≤ 5 % cross-overs on any of the 3 assay cutoffs.

Any modification of the instrument as set forth in this labeling requires validation by the laboratory.

References

1. Karch SB, ed. Drug Abuse Handbook. Boca Raton, FL: CRC Press LLC 1998.
1. Borenstein M. Central Nervous System Stimulants. In: Troy D. ed. Remington: The Science and Practice of Pharmacy. 21st ed. Baltimore, MD: Lippincott Williams & Wilkins Co 2005:1551.
1. NIDA Research Report Methamphetamine Abuse and Addiction: NIH Publication No. 06-4210. National Institute on Drug Abuse 6001 Executive Blvd., Room 5213, Bethesda, MD 20892-9561.
1. Baselt RC. Disposition of Toxic Drugs and Chemicals in Man. 7th ed. Foster City, CA: Biomedical Publications 2004:67.
1. Armbruster DA, Schwarzhoff RH, Pierce BL, Hubster EC. Method comparison of EMIT II and ONLINE with RIA for drug screening. J Forensic Sci 1993;38:1326-1341.
1. Armbruster DA, Schwarzhoff RH, Hubster EC, Liserio MK. Enzyme immunoassay, kinetic microparticle immunoassay, radioimmunoassay, and fluorescence polarization immunoassay compared for drugs-of-abuse screening. Clin Chem 1993;39:2137-2146.
1. Rouse S, Motter K, McNally A, et al. An Abuscreen OnLine Immunoassay for the Detection of Amphetamine in Urine on the COBAS MIRA Automated Analyzer. Clin Chem 1991;37(6):995. Abstract.
1. Toxicology and Drug Testing in the Clinical Laboratory; Approved Guideline. 2nd ed. (C52-A2). Wayne, PA: Clinical and Laboratory Standards Institute 2007;27:33.
1. Mandatory Guidelines for Federal Workplace Drug Testing Programs (Revised Specimen Validity Testing). Fed Regist 2004;69:19643-19673.
1. Data on file at Roche Diagnostics.

COBAS INTEGRA, COBAS C, and PRECISET are trademarks of Roche. Other brands are trademarks of their respective holders.

C 2010, Roche Diagnostics

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Image /page/30/Picture/1 description: The image shows the logo for the Department of Health & Human Services USA. The logo consists of a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" arranged around the top half of the circle. Inside the circle is a stylized emblem featuring a symbol that resembles an abstract eagle or bird-like figure with three distinct, curved lines forming the body and wings.

Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center - WO66-0609 Silver Spring, MD 20993-0002

Jan 2 8 2010

Roche Diagnostics c/o Mrs. Michelle Neff 9115 Hague Road Indianapolis, IN 46250

Re: K093664

Trade/Device Name: Amphetamines II Assay Regulation Number: 21 CFR §862.3100 Regulation Name: Amphetamine Test System Regulatory Class: Class II Product Code: DKZ, LAF Dated: July 2, 2010 Received: July 6, 2010

Dear Mrs. Neff:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include reguirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration.

If your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

31

Page 2

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding postmarket surveillance, please contact CDRH's Office of Surveillance and Biometric's (OSB's) Division of Postmarket Surveillance at (301) 796-5760. For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or ( 301 ) 796-5680 or at its Internet address http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Signature

Courtney C. Harper, Ph.D. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

32

Indication for Use

510(k) Number (if known):

K093664

JUL 2 8 2010

Device Name: Amphetamines II Indication For 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 a-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.1 Clinical consideration and professional judgement should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

Prescription Use XXX (21 CFR Part 801 Subpart D) And/Or

Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K093664