ONLINE DAT Benzodiazepines II

K221765 · Roche Diagnostics · JXM · Dec 23, 2022 · Clinical Toxicology

Device Facts

• Record ID: K221765
• Device Name: ONLINE DAT Benzodiazepines II
• Applicant: Roche Diagnostics
• Product Code: JXM · Clinical Toxicology
• Decision Date: Dec 23, 2022
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3170
• Device Class: Class 2

Intended Use

Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on cobas c systems at cutoff concentrations of 100 ng/mL, 200 ng/mL, and 300 ng/mL. Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program. Semiquantitative assays are intended to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas chromatography/mass spectrometry (GC-MS), or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Benzodiazepines II provides only a preliminary analytical test result. A more specific alternical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC-MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

Device Story

In vitro diagnostic immunoassay for benzodiazepine detection in human urine. Uses kinetic interaction of microparticles in solution (KIMS) on automated cobas c systems. Reagents contain monoclonal/polyclonal antibodies, substrate, and enzyme-labeled drug conjugate. Principle: drug in sample competes with particle-bound drug for antibody; inhibition of particle aggregation measured photometrically via turbidity. Presence of p-glucuronidase enhances cross-reactivity to glucuronidated metabolites. Used in clinical laboratories by technicians/pathologists. Output: qualitative (positive/negative) or semiquantitative (ng/mL) results. Preliminary results require confirmation by GC-MS or LC-MS/MS. Clinical decision-making relies on professional judgment alongside test results. Benefits: rapid screening for drug abuse assessment.

Clinical Evidence

Bench testing only. Precision (CLSI EP5-A3), linearity (CLSI EP6-A), analytical specificity, and interference studies performed. Method comparison study evaluated 137 samples against LC-MS/MS confirmation. Results showed 100% agreement for negative and positive clinical samples at 100, 200, and 300 ng/mL cutoffs. Stability testing confirmed 12-week onboard reagent stability.

Technological Characteristics

KIMS (Kinetic interaction of microparticles in solution) assay. Photometric turbidity measurement. Two liquid reagents: antibody buffer and conjugated drug microparticles. Includes p-glucuronidase for enzymatic cleavage of glucuronidated metabolites. Automated platform (cobas c 501). Storage 2-8°C.

Indications for Use

Indicated for qualitative and semiquantitative detection of benzodiazepines in human urine for clinical laboratory use on cobas c systems. Cutoff concentrations: 100, 200, and 300 ng/mL. Provides preliminary analytical results; requires confirmation by GC-MS or LC-MS/MS.

Regulatory Classification

Identification

A benzodiazepine test system is a device intended to measure any of the benzodiazepine compounds, sedative and hypnotic drugs, in blood, plasma, and urine. The benzodiazepine compounds include chlordiazepoxide, diazepam, oxazepam, chlorzepate, flurazepam, and nitrazepam. Measurements obtained by this device are used in the diagnosis and treatment of benzodiazepine use or overdose and in monitoring levels of benzodiazepines to ensure appropriate therapy.

Special Controls

*Classification.* Class II (special controls). A benzodiazepine 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).

Submission Summary (Full Text)

{0}------------------------------------------------

Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: a symbol on the left and the FDA name on the right. The symbol on the left is a stylized image of a human figure, while the FDA name on the right is written in blue letters. The words "U.S. FOOD & DRUG ADMINISTRATION" are written in a clear, sans-serif font.

December 23, 2022

Roche Diagnostics Khoa Tran Regulatory Affairs Program Manager 9115 Hague Road Indianapolis, IN 46250

### Re: K221765

Trade/Device Name: ONLINE DAT Benzodiazepines II Regulation Number: 21 CFR 862.3170 Regulation Name: Benzodiazepine test system Regulatory Class: Class II Product Code: JXM Dated: June 15, 2022 Received: June 17, 2022

### Dear Khoa Tran:

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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. 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 Part

{1}------------------------------------------------

801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4. Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Digitally signed by Paula_________________________________________________________________________________________________________________________________________________________________________ Paula Caposino -S Caposino -S Date: 2022.12.23

Paula Caposino, Ph.D. Acting Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Ouality Center for Devices and Radiological Health

Enclosure

{2}------------------------------------------------

# Indications for Use

510(k) Number (if known) K221765

Device Name ONLINE DAT Benzodiazepines II

### Indications for Use (Describe)

Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on cobas c systems at cutoff concentrations of 100 ng/mL, and 300 ng/mL. Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program.

Semiquantitative assays are intended to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas chromatography/mass spectrometry (GC-MS), or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS).

Benzodiazepines II provides only a preliminary analytical test result. A more specific alternical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC-MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

| Type of Use (Select one or both, as applicable) | |
|------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------|
| <div> <span> <span style="font-size:16px">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) </span> </div> | <div> <span> <span style="font-size:16px">☐</span> Over-The-Counter Use (21 CFR 801 Subpart C) </span> </div> |

### CONTINUE ON A SEPARATE PAGE IF NEEDED.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

### *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.*

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

> Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

{3}------------------------------------------------

# ONLINE DAT Benzodiazepines II K221765 - 510(k) Summary

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.

In accordance with 21 CFR 807.87. Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).

The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the ONLINE DAT Benzodiazepines II

{4}------------------------------------------------

| Submitter Name | Roche Diagnostics |
|--------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Address | 9115 Hague Road<br>P.O. Box 50416<br>Indianapolis, IN 46250-0457 |
| Contact | Khoa Tran<br>Phone: [phone]<br>Email: [e-mail]<br><br>Secondary Contact Name: Leslie Patterson<br>Phone: [phone]<br>Email: [e-mail] |
| Date Prepared | June 6, 2022 |
| Proprietary Name | ONLINE DAT Benzodiazepines II |
| Common Name | Benzodiazepines Enzyme Immunoassay |
| Classification Name and<br>Panel | Benzodiazepine Test System, 91 – Toxicology |
| Product Codes,<br>Regulation Numbers | JXM, Class II, 21 CFR 862. 3170 |
| Predicate Devices | ONLINE DAT Benzodiazepines Plus |
| Establishment Registration | Roche Diagnostics GmbH Mannheim, Germany: 9610126<br>Roche Diagnostics GmBH Penzberg, Germany: 9610529<br>Roche Diagnostics Indianapolis, IN United States: 1823260. |

{5}------------------------------------------------

#### 1. DEVICE DESCRIPTION

The Benzodiazepines II assay is an in vitro diagnostic test for the qualitative and semi-quantitative detection of benzodiazepines in human urine on automated clinical chemistry analyzers at cutoff concentrations of 100 ng/mL and 300 ng/mL. The semi quantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program.

#### 1.1. ONLINE DAT Benzodiazepines II

The device consists of two wet reagents which contain the key components of the immunoassay: monoclonal/ polyclonal antibody against the drug, substrate, and enzyme-labeled drug (conjugate).

#### 2. INDICATIONS FOR USE

Benzodiazepines II (BNZ2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of benzodiazepines in human urine on cobas c systems at cutoff concentrations of 100 ng/mL, 200 ng/mL, and 300 ng/mL.

Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program. Semiquantitative assays are intended to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas chromatography/mass spectrometry (GC-MS), or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS).

Benzodiazepines II provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC-MS) or Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug of abuse test result, particularly when preliminary positive results are used.

{6}------------------------------------------------

#### TECHNOLOGICAL CHARACTERISTICS 3.

The assay is based on the kinetic interaction of microparticles in a solution (KIMS) as measured by changes in light transmission. In the absence of sample drug, free antibody binds to drugmicroparticle conjugates causing the formation of particle aggregates that are photometrically detected by turbidity measurements. As the aggregation reaction proceeds in the absence of sample drug, the absorbance increases.

When a urine sample contains the drug in question, this drug competes with the particle-bound drug derivative for free antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.

The presence of p-glucuronidase enzyme enhances the Benzodiazepines II assay cross-reactivity to some of the glucuronidated metabolites. Enzymatic cleavage makes the benzodiazepine part of the glucuronides more accessible for the antibody.

The assay consists of two liquid reagents. Benzodiazepines antibody buffer and conjugated benzodiazepine derivative microparticles.

The following table compare the ONLINE DAT Benzodiazepines II with its predicate device, ONLINE DAT Benzodiazepines Plus (k043327).

{7}------------------------------------------------

| Feature | Candidate Device<br>ONLINE DAT Benzodiazepines<br>II | Predicate Device<br>ONLINE DAT Benzodiazepines<br>Plus (k043327) |
|---------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | Benzodiazepines II (BNZ2) is an<br>in vitro diagnostic test for the<br>qualitative and semiquantitative<br>detection of benzodiazepines in<br>human urine on Roche/Hitachi<br>cobas c systems at cutoff<br>concentrations of 100 ng/mL, 200<br>ng/mL, and 300 ng/mL.<br>Semiquantitative test results may<br>be obtained that permit<br>laboratories to assess assay<br>performance as part of a quality<br>control program.<br>Benzodiazepines II provides only a<br>preliminary analytical test result. A<br>more specific alternate chemical<br>method must be used in order to<br>obtain a confirmed analytical<br>result. Gas chromatography/mass<br>spectrometry (GC-MS) or Liquid<br>Chromatography coupled with<br>Tandem Mass Spectrometry<br>(LC-MS/MS) is the preferred<br>confirmatory method. Clinical<br>consideration and professional<br>judgment should be applied to any<br>drug of abuse test result,<br>particularly when preliminary<br>positive results are used. | The ONLINE DAT Benzodiazepines<br>Plus is an in vitro diagnostic test for<br>the qualitative and semi-quantitative<br>detection of benzodiazepines in<br>human urine on automated clinical<br>chemistry analyzers at cutoff<br>concentrations of 100 ng/mL, 200<br>ng/mL, and 300 ng/mL. Semi-<br>quantitative test results may be<br>obtained that permit laboratories to<br>assess assay performance as part of a<br>quality control program. |
| Detection Method | KIMS, Kinetic interaction of<br>microparticles in a solution | Same |
| Instrument Platform | cobas c 501 | Mod P |
| Test Matrix | Urine | Same |
| Measured Analyte | Benzodiazepine and its<br>metabolites | Benzodiazepine |
| Cutoff Levels | 100 ng/mL, 200 ng/mL and 300<br>ng/mL | Same |
| Reagent Stability | 2-8 °C until expiration date | Same |

### Technical Characteristics Comparison Table between ONLINE DAT Table 1: Benzodiazepines II and ONLINE DAT Benzodiazepines Plus

{8}------------------------------------------------

#### NON-CLINICAL PERFORMANCE EVALUATION 4.

The following performance data are provided in support of the substantial equivalence determination:

- Precision according to CLSI EP5-A3 .
- Linearity according to CLSI EP6-A .
- Analytical Specificity/Cross-Reactivity .
- Endogenous Interferences .
- Interference Drugs .
- Interference Testing of Specific Gravity and pH .
- Method Comparison to Predicate .
- Stability .

All performance specifications were met.

#### 4.1. Precision

#### Repeatability and Intermediate Precision 4.1.1.

The precision study was performed using CLSI Guideline EP05-A3 as a guideline. Precision experiments were conducted using one reagent lot on one cobas c 501 instrument. Testing was carried out for 21 days with two runs per day, and two replicates per run in both Qualitative and Semi-quantitative modes, giving a total of 84 determinants (n = 84). Drug-free negative urine was spiked with Oxazepam to final concentrations of -50%, below cutoff and +25%, +50% above cutoff.

| 100 ng/mL SQ<br>Repeatability | Mean (ng/mL) | SD (ng/mL) | CV (%) |
|-------------------------------|--------------|------------|--------|
| Urine -50% | 45.0 | 2.56 | 5.7 |
| Urine -25% | 68.8 | 2.44 | 3.6 |

{9}------------------------------------------------

| DAT2N<br>(control 75 ng/mL) | 78.8 | 1.99 | 2.5 |
|------------------------------|--------------|------------|--------|
| Cutoff Urine | 99.7 | 2.38 | 2.4 |
| Urine +25% | 123 | 2.43 | 2.0 |
| DAT2P<br>(control 125 ng/mL) | 127 | 2.24 | 1.8 |
| Urine +50% | 146 | 2.59 | 1.8 |
| Intermediate<br>Precision | Mean (ng/mL) | SD (ng/mL) | CV (%) |
| Urine -50% | 45.0 | 2.79 | 6.2 |
| Urine -25% | 68.8 | 2.65 | 3.9 |
| DAT2N<br>(control 75 ng/mL) | 78.8 | 2.79 | 3.5 |
| Cutoff Urine | 99.7 | 2.63 | 2.6 |
| Urine +25% | 123 | 2.92 | 2.4 |
| DAT2P<br>(control 125 ng/mL) | 127 | 3.21 | 2.5 |
| Urine +50% | 146 | 2.94 | 2.0 |

| 100 ng/mL<br>Qualitative | Number Tested | Negative/Positive | Confidence Level |
|-----------------------------|---------------|-------------------|-------------------------|
| Urine -50% | 84 | 84/0 | > 95 % negative reading |
| Urine -25% | 84 | 84/0 | > 95 % negative reading |
| DAT2N<br>(control 75 ng/mL) | 84 | 84/0 | > 95 % negative reading |

{10}------------------------------------------------

| Cutoff Urine | 84 | 73/11 | N/A |
|------------------------------|----|-------|-------------------------|
| Urine +25% | 84 | 0/84 | > 95 % positive reading |
| DAT2P<br>(control 125 ng/mL) | 84 | 0/84 | > 95 % positive reading |
| Urine +50% | 84 | 0/84 | > 95 % positive reading |

| Repeatability<br>200 ng/mL SQ | Mean (ng/mL) | SD (ng/mL) | CV (%) |
|-------------------------------|--------------|------------|--------|
| Urine -50% | 98.2 | 2.50 | 2.5 |
| Urine -25% | 146 | 2.51 | 1.7 |
| DAT3N<br>(control 150 ng/mL) | 150 | 1.91 | 1.3 |
| Cutoff Urine | 199 | 2.48 | 1.2 |
| Urine +25% | 242 | 3.15 | 1.3 |
| DAT3P<br>(control 250 ng/mL) | 248 | 2.67 | 1.1 |
| Urine +50% | 279 | 2.34 | 0.8 |
| Intermediate<br>Precision | Mean (ng/mL) | SD (ng/mL) | CV (%) |
| Urine -50% | 98.2 | 3.09 | 3.1 |
| Urine -25% | 146 | 2.87 | 2.0 |
| DAT3N<br>(control 150 ng/mL) | 150 | 3.49 | 2.3 |
| Cutoff Urine | 199 | 3.33 | 1.7 |
| Urine +25% | 242 | 3.72 | 1.5 |

{11}------------------------------------------------

| DAT3P<br>(control 250 ng/mL) | 248 | 5.68 | 2.3 |
|------------------------------|-----|------|-----|
| Urine +50% | 279 | 4.88 | 1.7 |

| 200 ng/mL<br>Qualitative | Number Tested | Negative/Positive | Confidence Level |
|------------------------------|---------------|-------------------|-------------------------|
| Urine -50% | 84 | 84/0 | > 95 % negative reading |
| Urine -25% | 84 | 84/0 | > 95 % negative reading |
| DAT3N<br>(control 150 ng/mL) | 84 | 84/0 | > 95 % negative reading |
| Cutoff Urine | 84 | 60/24 | N/A |
| Urine +25% | 84 | 0/84 | > 95 % positive reading |
| DAT3P<br>(control 250 ng/mL) | 84 | 0/84 | > 95 % positive reading |
| Urine +50% | 84 | 0/84 | > 95 % positive reading |

| 300 ng/mL SQ<br>Repeatability | Mean (ng/mL) | SD (ng/mL) | CV (%) |
|-------------------------------|--------------|------------|--------|
| Urine -50% | 151 | 4.41 | 2.9 |
| Urine -25% | 211 | 3.99 | 1.9 |
| DAT1N<br>(control 225 ng/mL) | 223 | 5.50 | 2.5 |
| Cutoff Urine | 276 | 4.17 | 1.5 |
| Urine +25% | 354 | 5.21 | 1.5 |

{12}------------------------------------------------

| DAT1P<br>(control 375 ng/mL) | 363 | 4.35 | 1.2 |
|------------------------------|--------------|------------|--------|
| Urine +50% | 432 | 5.14 | 1.2 |
| Intermediate<br>Precision | Mean (ng/mL) | SD (ng/mL) | CV (%) |
| Urine -50% | 151 | 5.31 | 3.5 |
| Urine -25% | 211 | 5.40 | 2.6 |
| DAT1N<br>(control 225 ng/mL) | 223 | 6.23 | 2.8 |
| Cutoff Urine | 276 | 6.07 | 2.2 |
| Urine +25% | 354 | 6.17 | 1.7 |
| DAT1P<br>(control 375 ng/mL) | 363 | 7.35 | 2.0 |
| Urine +50% | 432 | 6.70 | 1.6 |

| 300 ng/mL<br>Qualitative | Number Tested | Negative/Positive | Confidence Level |
|------------------------------|---------------|-------------------|-------------------------|
| Urine -50% | 84 | 84/0 | > 95 % negative reading |
| Urine -25% | 84 | 84/0 | > 95 % negative reading |
| DAT1N<br>(control 225 ng/mL) | 84 | 84/0 | > 95 % negative reading |
| Cutoff Urine | 84 | 83/1 | N/A |
| Urine +25% | 84 | 0/84 | > 95 % positive reading |
| DAT1P<br>(control 375 ng/mL) | 84 | 0/84 | > 95 % positive reading |

{13}------------------------------------------------

| Urine +50% | 84 | 0/84 | > 95 % positive reading |
|------------|----|------|-------------------------|
|------------|----|------|-------------------------|

#### Analytical Recovery and Linearity 4.2.

The recovery study was evaluated on a single cobas c 501 in according to CLSI guideline EP06-A. The study protocol consisted of three reagent lots, the total number of samples was 17 levels per cutoff and ran in triplicate. The recovery study experiment was conducted using three reagent lots on one cobas c 501 instrument. Two series of samples were prepared for each cutoff to support that the recovery performance of the device is acceptable over the whole range between the lowest and the highest calibrator.

For each sample, the percentage recovery was calculated as the percent of the three results with regard to the target value. The average percent recovery is summarized in the table below.

| 100 Cutoff | Lot 1 | | Lot 2 | | Lot 3 | |
|-------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|
| Target<br>(ng/mL) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) |
| 0 | 1 | N/A | 0 | N/A | 4 | N/A |
| 25 | 27 | 108 | 29 | 116 | 29 | 115 |
| 50 | 54 | 107 | 53 | 105 | 52 | 105 |
| 75 | 76 | 102 | 76 | 101 | 75 | 100 |
| 100 | 99 | 99 | 99 | 99 | 100 | 100 |
| 125 | 122 | 98 | 122 | 98 | 121 | 97 |
| 150 | 146 | 97 | 145 | 97 | 148 | 99 |
| 175 | 169 | 97 | 168 | 96 | 172 | 98 |
| 200 | 188 | 94 | 189 | 95 | 193 | 97 |

Results for 0-200% of 100 ng/mL cutoff

{14}------------------------------------------------

| 100 Cutoff | Lot 1 | | Lot 2 | | Lot 3 | |
|-------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|
| Target<br>(ng/mL) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) |
| 0 | 1 | N/A | 0 | N/A | 4 | N/A |
| 100 | 98 | 98 | 100 | 100 | 101 | 101 |
| 200 | 191 | 96 | 191 | 96 | 195 | 97 |
| 300 | 288 | 96 | 291 | 97 | 295 | 98 |
| 400 | 399 | 100 | 399 | 100 | 404 | 101 |
| 500 | 517 | 103 | 524 | 105 | 522 | 104 |
| 600 | 638 | 106 | 644 | 107 | 638 | 106 |
| 700 | 753 | 108 | 763 | 109 | 752 | 107 |
| 800 | 842 | 105 | 856 | 107 | 856 | 107 |
| 900 | 921 | 102 | 928 | 103 | 921 | 102 |
| 1000 | 982 | 98 | 978 | 98 | 988 | 99 |

Results for 0-1000% of 100 ng/mL cutoff

Results for 0-200% of 200 ng/mL cutoff

| 200 Cutoff | Lot 1 | | Lot 2 | | Lot 3 | |
|-------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|
| Target<br>(ng/mL) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) |
| 0 | 1 | N/A | 0 | N/A | 1 | N/A |
| 50 | 52 | 104 | 54 | 107 | 52 | 103 |
| 100 | 97 | 97 | 100 | 100 | 99 | 99 |
| 150 | 142 | 95 | 144 | 96 | 143 | 96 |
| 200 | 191 | 95 | 190 | 95 | 192 | 96 |
| 250 | 243 | 97 | 238 | 95 | 242 | 97 |

{15}------------------------------------------------

| 300 | 292 | 97 | 289 | 96 | 295 | 98 |
|-----|-----|-----|-----|-----|-----|-----|
| 350 | 351 | 100 | 347 | 99 | 349 | 100 |
| 400 | 408 | 102 | 403 | 101 | 406 | 101 |

Results for 0-1000% of 200 ng/mL cutoff

| 200 Cutoff<br>Target<br>(ng/mL) | Mean<br>(ng/mL) | Lot 1<br>Recovery<br>(%) | Mean<br>(ng/mL) | Lot 2<br>Recovery<br>(%) | Mean<br>(ng/mL) | Lot 3<br>Recovery<br>(%) |
|---------------------------------|-----------------|--------------------------|-----------------|--------------------------|-----------------|--------------------------|
| 0 | 1 | N/A | 0 | N/A | 1 | N/A |
| 100 | 99 | 99 | 100 | 100 | 99 | 99 |
| 200 | 193 | 96 | 191 | 95 | 193 | 97 |
| 300 | 293 | 98 | 289 | 96 | 296 | 99 |
| 400 | 404 | 101 | 399 | 100 | 399 | 100 |
| 500 | 526 | 105 | 518 | 104 | 517 | 103 |
| 600 | 646 | 108 | 643 | 107 | 634 | 106 |
| 700 | 760 | 109 | 761 | 109 | 751 | 107 |
| 800 | 857 | 107 | 852 | 107 | 850 | 106 |
| 900 | 927 | 103 | 927 | 103 | 922 | 102 |
| 1000 | 986 | 99 | 982 | 98 | 986 | 99 |

Results for 0-200% of 300 ng/mL cutoff

| 300 Cutoff<br>Target<br>(ng/mL) | Lot 1 | | Lot 2 | | Lot 3 | |
|---------------------------------|-----------------|-----------------|-----------------|-----------------|-----------------|-----------------|
| | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) | Mean<br>(ng/mL) | Recovery<br>(%) |
| 0 | 0 | N/A | 0 | N/A | 9 | N/A |
| 75 | 89 | 119 | 84 | 112 | 87 | 116 |
| 150 | 155 | 104 | 154 | 103 | 160 | 106 |

{16}------------------------------------------------

| 225 | 216 | 96 | 220 | 98 | 224 | 99 |
|-----|-----|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------|-----------|
| 300 | 281 | 94 | 283 | 94 | 300 | 100 |
| 375 | 349 | તે તે જેવી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચ | 348 | ਰੇਤੇ | 351 | 94 |
| 450 | 416 | 92 | 418 | તે તે જેવી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચ | 432 | તે તે રેણ |
| 525 | 482 | 92 | 487 | ਰੇਤੋ | 491 | 94 |
| 600 | 550 | 92 | 556 | તે તે જેવી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામમાં પ્રાથમિક શાળા, પંચ | ર્સા | તે તે ઉ |

Results for 0-1000% of 300 ng/mL cutoff

| 300 Cutoff<br>Target<br>(ng/mL) | Mean<br>(ng/mL) | Lot 1<br>Recovery<br>(%) | Mean<br>(ng/mL) | Lot 2<br>Recovery<br>(%) | Mean<br>(ng/mL) | Lot 3<br>Recovery<br>(%) |
|---------------------------------|-----------------|--------------------------|-----------------|--------------------------|-----------------|--------------------------|
| 0 | 0 | N/A | 0 | N/A | 9 | N/A |
| 300 | 284 | 95 | 281 | 94 | 288 | 96 |
| 600 | 554 | 92 | 557 | 93 | 561 | 93 |
| 900 | 869 | 97 | 888 | 99 | 869 | 97 |
| 1200 | 1253 | 104 | 1275 | 106 | 1228 | 102 |
| 1500 | 1687 | 112 | 1756 | 117 | 1639 | 109 |
| 1800 | 2096 | 116 | 2143 | 119 | 2069 | 115 |
| 2100 | 2422 | 115 | 2489 | 119 | 2424 | 115 |
| 2400 | 2679 | 112 | 2711 | 113 | 2674 | 111 |
| 2700 | 2837 | 105 | 2833 | 105 | 2831 | 105 |
| 3000 | 2903 | 97 | 2862 | 95 | 2907 | 97 |

{17}------------------------------------------------

#### Analytical Specificity/Cross-Reactivity 4.3.

The determination of cross reactivity by common benzodiazepines was conducted on one cobas c 501, using one reagent lot. Concentration series were prepared for each cross reactant by spiking drug free urine. The percent cross-reactivity was calculated from the ratio of the cutoff concentration and the calculated equivalent concentration of the cross reactant. The study was conducted for the semi-quantitative application. Results are summarized below:

| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
|------------------------------------|----------------------|----------------------|
| Nordiazepam | 101 | 99% |
| Alprazolam | 92 | 109% |
| Diazepam | 90 | 111% |
| Lorazepam | 105 | 95% |
| Lorazepam glucuronide | 178 | 56% |
| Oxazepam | 89 | 113% |
| Oxazepam glucuronide | 135 | 74% |
| Temazepam | 94 | 106% |
| Temazepam glucuronide | 160 | 63% |
| 3-Hydroxybromazepam | 153 | 66% |
| 3-Hydroxyflubromazepam | 130 | 77% |
| 3-Hydroxyflunitrazepam | 118 | 85% |
| 4-Hydroxyalprazolam | 63 | 160% |
| 4-Hydroxytriazolam | 98 | 102% |
| 7-Acetamidonitrazepam | 21421 | 0.5% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| 7-Aminoclonazepam | 124 | 80% |
| 7-Aminoflunitrazepam | 109 | 91% |
| 7-Aminonimetazepam | 87 | 115% |
| 7-Aminonitrazepam | 73 | 137% |
| Bentazepam | 128 | 78% |
| Bromazepam | 76 | 132% |
| Brotiazolam | 138 | 73% |
| Chlordiazepoxide | 109 | 92% |
| Clobazam | 95 | 106% |
| Clonazepam | 103 | 97% |
| Clonazolam | 110 | 91% |
| Clorazepate | 189 | 53% |
| Delorazepam | 109 | 92% |
| Demoxepam | 76 | 131% |
| Desalkylflurazepam | 97 | 103% |
| Deschloretizolam | 81 | 124% |
| Desmethylchlordiazepoxide | 108 | 93% |
| Desmethylflunitrazepam | 100 | 100% |
| Desmethylmedazepam | 168 | 59% |
| Diclazepam | 99 | 101% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Didesethylflurazepam | 116 | 86% |
| Estazolam | 88 | 114% |
| Etizolam | 118 | 85% |
| Flubromazepam | 132 | 76% |
| Flubromazolam | 105 | 95% |
| Flunitrazepam | 113 | 88% |
| Flurazepam | 161 | 62% |
| Halazepam | 132 | 76% |
| Hydroxyethylflurazepam | 103 | 97% |
| Lormetazepam | 107 | 94% |
| Meclonazepam | 123 | 82% |
| Medazepam | 138 | 72% |
| Midazolam | 106 | 95% |
| Nifoxipam | 129 | 78% |
| Nimetazepam | 99 | 101% |
| Nitrazepam | 96 | 105% |
| Phenazepam | 124 | 81% |
| Pinazepam | 110 | 91% |
| Prazepam | 124 | 80% |
| Pyrazolam | 103 | 97% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Tetrazepam | 116 | 86% |
| Triazolam | 103 | 97% |
| α-Hydroxyalprazolam | 85 | 118% |
| α-Hydroxyalprazolam<br>glucuronide | 190 | 53% |
| α-Hydroxymidazolam | 103 | 97% |
| α-Hydroxymidazolam<br>glucuronide | 179 | 56% |
| α-Hydroxytriazolam | 96 | 105% |

Cross Reactivity of Benzodiazepines and Metabolites for 100 ng/mL Cutoff

{18}------------------------------------------------

{19}------------------------------------------------

{20}------------------------------------------------

Cross Reactivity of Benzodiazepines and Benzodiazepines Metabolites for 200 ng/mL

| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
|------------------------------------|----------------------|----------------------|
| Nordiazepam | 198 | 101% |
| Alprazolam | 174 | 115% |
| Diazepam | 175 | 115% |
| Lorazepam | 208 | 96% |
| Lorazepam glucuronide | 360 | 56% |
| Oxazepam | 193 | 104% |
| Oxazepam glucuronide | 282 | 71% |
| Temazepam | 193 | 103% |
| Temazepam glucuronide | 318 | 63% |
| 3-Hydroxybromazepam | 371 | 54% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| 3-Hydroxyflubromazepam | 256 | 78% |
| 3-Hydroxyflunitrazepam | 259 | 77% |
| 4-Hydroxyalprazolam | 119 | 169% |
| 4-Hydroxytriazolam | 203 | 99% |
| 7-Acetamidonitrazepam | 42005 | 0.5% |
| 7-Aminoclonazepam | 283 | 71% |
| 7-Aminoflunitrazepam | 219 | 91% |
| 7-Aminonimetazepam | 213 | 94% |
| 7-Aminonitrazepam | 177 | 113% |
| Bentazepam | 288 | 69% |
| Bromazepam | 154 | 130% |
| Brotiazolam | 265 | 76% |
| Chlordiazepoxide | 282 | 71% |
| Clobazam | 185 | 108% |
| Clonazepam | 208 | 96% |
| Clonazolam | 226 | 89% |
| Clorazepate | 396 | 50% |
| Delorazepam | 217 | 92% |
| Demoxepam | 182 | 110% |
| Desalkylflurazepam | 194 | 103% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Deschloretizolam | 159 | 126% |
| Desmethylchlordiazepoxide | 304 | 66% |
| Desmethylflunitrazepam | 196 | 102% |
| Desmethylmedazepam | 399 | 50% |
| Diclazepam | 202 | 99% |
| Didesethylflurazepam | 231 | 87% |
| Estazolam | 168 | 119% |
| Etizolam | 236 | 85% |
| Flubromazepam | 266 | 75% |
| Flubromazolam | 203 | 98% |
| Flunitrazepam | 212 | 94% |
| Flurazepam | 312 | 64% |
| Halazepam | 269 | 74% |
| Hydroxyethylflurazepam | 206 | 97% |
| Lormetazepam | 214 | 94% |
| Meclonazepam | 306 | 65% |
| Medazepam | 279 | 72% |
| Midazolam | 206 | 97% |
| Nifoxipam | 310 | 64% |
| Nimetazepam | 201 | 99% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Nitrazepam | 188 | 106% |
| Phenazepam | 252 | 79% |
| Pinazepam | 213 | 94% |
| Prazepam | 240 | 83% |
| Pyrazolam | 203 | 99% |
| Tetrazepam | 233 | 86% |
| Triazolam | 207 | 96% |
| α-Hydroxyalprazolam | 168 | 119% |
| α-Hydroxyalprazolam<br>glucuronide | 399 | 50% |
| α-Hydroxymidazolam | 201 | 100% |
| α-Hydroxymidazolam<br>glucuronide | 372 | 54% |
| α-Hydroxytriazolam | 198 | 101% |

{21}------------------------------------------------

{22}------------------------------------------------

{23}------------------------------------------------

Cross Reactivity of Benzodiazepines and Benzodiazepines Metabolites for 300 ng/mL Cutoffs

| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
|------------------------------------|----------------------|----------------------|
| Nordiazepam | 305 | 98% |
| Alprazolam | 275 | 109% |
| Diazepam | 273 | 110% |
| Lorazepam | 323 | 93% |
| Lorazepam glucuronide | 519 | 58% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Oxazepam | 275 | 109% |
| Oxazepam glucuronide | 442 | 68% |
| Temazepam | 272 | 110% |
| Temazepam glucuronide | 496 | 61% |
| 3-Hydroxybromazepam | 504 | 59% |
| 3-Hydroxyflubromazepam | 401 | 75% |
| 3-Hydroxyflunitrazepam | 399 | 75% |
| 4-Hydroxyalprazolam | 192 | 156% |
| 4-Hydroxytriazolam | 319 | 94% |
| 7-Acetamidonitrazepam | 93148 | 0.3% |
| 7-Aminoclonazepam | 390 | 77% |
| 7-Aminoflunitrazepam | 343 | 87% |
| 7-Aminonimetazepam | 283 | 106% |
| 7-Aminonitrazepam | 230 | 131% |
| Bentazepam | 393 | 76% |
| Bromazepam | 229 | 131% |
| Brotiazolam | 407 | 74% |
| Chlordiazepoxide | 374 | 80% |
| Clobazam | 277 | 108% |
| Clonazepam | 317 | 95% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Clonazolam | 338 | 89% |
| Clorazepate | 601 | 50% |
| Delorazepam | 334 | 90% |
| Demoxepam | 256 | 117% |
| Desalkylflurazepam | 307 | 98% |
| Deschloretizolam | 257 | 117% |
| Desmethylchlordiazepoxide | 370 | 81% |
| Desmethylflunitrazepam | 301 | 100% |
| Desmethylmedazepam | 549 | 55% |
| Diclazepam | 317 | 95% |
| Didesethylflurazepam | 352 | 85% |
| Estazolam | 270 | 111% |
| Etizolam | 362 | 83% |
| Flubromazepam | 435 | 69% |
| Flubromazolam | 327 | 92% |
| Flunitrazepam | 329 | 91% |
| Flurazepam | 487 | 62% |
| Halazepam | 406 | 74% |
| Hydroxyethylflurazepam | 324 | 92% |
| Lormetazepam | 328 | 92% |
| Benzodiazepine and<br>metabolites | Concentration Tested | Cross-reactivity (%) |
| Meclonazepam | 409 | 73% |
| Medazepam | 394 | 76% |
| Midazolam | 332 | 90% |
| Nifoxipam | 412 | 73% |
| Nimetazepam | 309 | 97% |
| Nitrazepam | 294 | 102% |
| Phenazepam | 382 | 78% |
| Pinazepam | 327 | 92% |
| Prazepam | 363 | 83% |
| Pyrazolam | 311 | 96% |
| Tetrazepam | 360 | 83% |
| Triazolam | 315 | 95% |
| α-Hydroxyalprazolam | 275 | 109% |
| α-Hydroxyalprazolam<br>glucuronide | 567 | 53% |
| α-Hydroxymidazolam | 326 | 92% |
| α-Hydroxymidazolam<br>glucuronide | 569 | 53% |
| α-Hydroxytriazolam | 312 | 96% |

{24}------------------------------------------------

{25}------------------------------------------------

{26}------------------------------------------------

#### Endogenous Interference Testing of Structurally Unrelated Compounds 4.4.

Interference from structurally unrelated compounds was evaluated with two sets of samples were prepared and measured for the analysis of each individual interferent. One set of the samples

{27}------------------------------------------------

contained the interferent in the presence of a benzodiazepine at both target concentrations (75% and 125% of the cutoff of the assay), and one set of the samples contained the interferent in the presence of a glucuronidated benzodiazepine (oxazepam-glucuronide) at both target concentrations (75% or 125% of the cutoff of the assay). Testing was performed in both qualitative and semiquantitative modes. The compounds listed in the table below did not cause any positive or negative interference at the concentrations shown:

| Compound Name | Concentration in presence of benzodiazepine (mg/dL) |
|-------------------------------|--------------------------------------------------------------|
| Acetone | 1000 |
| Ascorbic Acid | 1500 |
| Calcium<br>(as CaCl2) | 133 |
| Citrate<br>(K3-Citrate x H2O) | 357 |
| Creatinine | 1000 |
| Ethanol | 1000 |
| Glucose | 7000 |
| Hemoglobin | 750 |
| Human Albumin | 250 |
| Human IgG | 110 |
| Magnesium<br>(MgCl2) | 238 |
| Oxalate<br>(Na2-Oxalate) | 20 |
| Phosphate<br>(NaH2PO4 x H2O) | 2028 |
| Sodium chloride | 5844 |
| Urea | 18000 |
| Compound Name | Concentration in<br>presence of<br>benzodiazepine<br>(mg/dL) |
| Uric Acid | 100 |
| Urobilinogen | 15 |

{28}------------------------------------------------

#### Interference Testing of Specific Gravity and pH 4.5.

Urine samples within a pH range from 4.0 to 9.0 and samples with specific gravities ranging from 1.001 to 1.034 containing benzodiazepine at the level of the negative control (75 ng/mL, 150 ng/mL or 225 ng/mL) and at the level of the positive control (125 ng/mL, 250 ng/mL or 375 ng/mL) corresponding respectively to the given cutoff (100 ng/mL, 200 ng/mL or 300 ng/mL) recovered properly in both semi-quantitative and qualitative modes.

#### Drug Interferences 4.6.

The drug interference study was conducted using one reagent lot on the cobas c 501. Samples spiked with benzodiazepine were measured in the presence of potentially interfering drugs. The target concentration for the benzodiazepine was 75% or the cutoff of the assay. For all samples three replicates were recorded with the semi-quantitative applications and the qualitative applications. The test results were checked for exceeding the cutoff value. The maximum drug concentration which doesn't interfere is reported and established as interferent claim. For the tested drugs and drug concentrations compare table below.

For Oxaprozin, the interference was additionally evaluated following an alternative protocol. In a drug-free matrix, the approximate quantity of Oxaprozin that is equivalent in assay reactivity to the 100 ng/mL, 200 ng/mL, and 300 ng/mL cutoff was determined to be 790 ng/mL, 3091 ng/mL and 3049 ng/mL respectively. This equals to a cross reactivity of 13% (at cutoff 100 ng/mL), 6% (at cutoff 200 ng/mL), and 10% (at cutoff 300 ng/mL).

When oxaprozin was added pooled human urine containing benzodiazepine at the level of the negative control (75 ng/mL, 150 ng/mL or 225 ng/mL), positive results were observed at >100 ng/mL, >200 ng/mL, and >300 ng/mL for the 100 ng/mL cutoff, 200 ng/mL cutoff, and 300 ng/mL

{29}------------------------------------------------

cutoff, respectively. Patient samples containing benzodiazepines in the presence of oxaprozin may yield falsely elevated results. Results should always be assessed in conjunction with the patient's medical history, clinical examinations, and other clinicopathological findings.

| Compounds | Concentration (ng/mL) | | |
|--------------------------------------------------------------|-----------------------|---------------------|---------------------|
| | Cutoff 100<br>ng/mL | Cutoff 200<br>ng/mL | Cutoff 300<br>ng/mL |
| Acetaminophen | 3000000 | 3000000 | 3000000 |
| Acetylsalicylic acid | 100000 | 100000 | 100000 |
| Amitryptyline | 100000 | 100000 | 100000 |
| Amobarbital | 100000 | 100000 | 100000 |
| d-Amphetamine | 100000 | 100000 | 100000 |
| 1-Amphetamine | 100000 | 100000 | 100000 |
| Ampicillin | 100000 | 100000 | 100000 |
| Aspartame | 100000 | 100000 | 100000 |
| Atropine | 100000 | 100000 | 100000 |
| Benzocaine | 100000 | 100000 | 100000 |
| Benzoylecgonine | 100000 | 100000 | 100000 |
| Benzphetamine | 100000 | 100000 | 100000 |
| Buspirone | 100000 | 100000 | 100000 |
| Butabarbital | 100000 | 100000 | 100000 |
| Ca-dobesilate | 1000000 | 1000000 | 1000000 |
| Caffeine | 100000 | 100000 | 100000 |
| Calcium hypochlorite | 100000 | 100000 | 100000 |
| Compounds | Concentration (ng/mL) | | |
| | Cutoff 100<br>ng/mL | Cutoff 200<br>ng/mL | Cutoff 300<br>ng/mL |
| Cannabidiol | 100000 | 100000 | 100000 |
| Captopril | 100000 | 100000 | 100000 |
| Cefoxitin | 2000000 | 4000000 | 6000000 |
| Chloroquine | 100000 | 100000 | 100000 |
| Chlorpheniramine | 40000 | 100000 | 100000 |
| Chlorpromazine | 100000 | 100000 | 100000 |
| Cocaine | 100000 | 100000 | 100000 |
| Codeine | 100000 | 100000 | 100000 |
| Desipramine | 100000 | 100000 | 100000 |
| Dextromethorphan | 100000 | 100000 | 100000 |
| Dextropropoxyphene (d-<br>Propoxyphene) | 100000 | 100000 | 100000 |
| Digoxin | 100000 | 100000 | 100000 |
| Diphenhydramine | 40000 | 100000 | 100000 |
| Doxepine | 100000 | 100000 | 100000 |
| Ecgonine | 100000 | 100000 | 100000 |
| Ecgonine methyl ester | 100000 | 100000 | 100000 |
| EDDP (2-Ethylidene-1,5-dimethyl-<br>3,3-diphenylpyrrolidine) | 25000 | 50000 | 75000 |
| EMDP (2-Ethyl-5-methyl-3,3-<br>diphenylpyrroline) | 25000 | 40000 | 80000 |
| Enalapril | 100000 | 100000 | 100000 |
| Compounds | Concentration (ng/mL) | | |
| | Cutoff 100<br>ng/mL | Cutoff 200<br>ng/mL | Cutoff 300<br>ng/mL |
| d-Ephedrine | 100000 | 100000 | 100000 |
| 1-Ephedrine | 100000 | 100000 | 100000 |
| Epinephrine | 100000 | 100000 | 100000 |
| Erythromycin | 100000 | 100000 | 100000 |
| Estriol | 100000 | 100000 | 100000 |
| Fenoprofen | 40000 | 100000 | 100000 |
| Flumazenil | 100000 | 100000 | 100000 |
| Furosemide | 100000 | 100000 | 100000 |
| Gentamicine sulfate | 400000 | 400000 | 400000 |
| Gentisic acid | 100000 | 100000 | 100000 |
| Glutethimide | 100000 | 100000 | 100000 |
| Guaiacol glycerol ether | 100000 | 100000 | 100000 |
| Hydrochlorothiazide | 100000 | 100000…