First Sign Drug of Abuse Dip Card Test, First Sign Drug of Abuse Cup Test

K150162 · W.H.P.M., Inc. · LAF · Feb 26, 2015 · Clinical Toxicology

Device Facts

• Record ID: K150162
• Device Name: First Sign Drug of Abuse Dip Card Test, First Sign Drug of Abuse Cup Test
• Applicant: W.H.P.M., Inc.
• Product Code: LAF · Clinical Toxicology
• Decision Date: Feb 26, 2015
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3610
• Device Class: Class 2

Intended Use

First Sign™ Drug of Abuse Tests are immunochromatographic assays for the qualitative determination of Oxazepam , Methamphetamine , and Morphine in human urine at cut-off concentrations of 300 ng/mL, 1000 ng/mL, and 2000 ng/mL, respectively. The tests are available in a Cup format and a Dip Card format. The tests may yield preliminary positive results even when prescription drug Oxazepam is ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for oxazepam in urine. The tests provide only preliminary test results. A more specific alternative chemical must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.

Device Story

First Sign™ Drug of Abuse Tests are lateral flow immunochromatographic assays for qualitative detection of Oxazepam, Methamphetamine, and Morphine in human urine. Available in Cup or Dip Card formats. Principle: competitive binding; target drugs in urine compete with drug-conjugate for limited antibody binding sites on particles. If drug concentration is below cut-off, antibody-coated particles bind to immobilized drug-conjugate, forming a visible colored line (negative result). If drug concentration exceeds cut-off, binding sites are saturated, preventing line formation (positive result). Used by lay users (OTC) or clinicians (prescription) for preliminary screening. Results are visually interpreted by the user. Positive results require confirmatory testing via GC/MS. Benefits include rapid, point-of-care screening for drug presence.

Clinical Evidence

Bench testing only. Performance validated via precision studies (25 days, 3 operators), interference testing with physiological substances, and specificity/cross-reactivity analysis. Method comparison studies (n=80 per drug) compared device results to GC/MS. A lay-user study (n=280 per drug) confirmed performance across diverse demographics and verified readability of instructions (Flesch-Kincaid Grade Level 7).

Technological Characteristics

Lateral flow immunochromatographic assay. Materials include monoclonal mouse antibodies. Form factors: Cup and Dip Card. Energy source: none (capillary action). Connectivity: none. Sterilization: not specified. Software: none.

Indications for Use

Indicated for the qualitative detection of Oxazepam, Methamphetamine, and Morphine in human urine for over-the-counter and prescription use. Not intended to distinguish between prescription use and abuse of Oxazepam. Provides preliminary results only; requires confirmation by GC/MS.

Regulatory Classification

Identification

A methamphetamine test system is a device intended to measure methamphetamine, a central nervous system stimulating drug, in serum, plasma, and urine. Measurements obtained by this device are used in the diagnosis and treatment of methamphetamine use or overdose.

Special Controls

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

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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized symbol that resembles a human figure in profile, with three overlapping heads.

February 26, 2015

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

W.H.P.M., INC. JOHN WAN 5358 IRWINDALE AVENUE IRWINDALE CA 91706

Re: K150162 Trade/Device Name: First Sign® Drug Of Abuse Cup Test First Sign® Drug Of Abuse Dip Card Test Regulation Number: 21 CFR 862.3610 Regulation Name: Methamphetamine test system Regulatory Class: II Product Code: LAF, JXM, DJG Dated: January 19, 2015 Received: January 26, 2015

Dear Mr. John Wan:

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 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. 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

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 Industry and Consumer Education at its toll-free number [phone] or [phone] or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Katherine Serrano -A

For : Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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### Indications for Use

510(k) Number (if known) k150162

Device Name First Sign® Drug of Abuse Cup Test First Sign® Drug of Abuse Dip Card Test

#### Indications for Use (Describe)

First Sign™ Drug of Abuse Tests are immunochromatographic assays for the qualitative determination of Oxazepam , Methamphetamine, and Morphine in human urine at cut-off concentrations of 300 ng/mL, and 2000 ng/mL, respectively. The tests are available in a Cup format and a Dip Card format.

The tests may yield preliminary positive results even when prescription drug Oxazepam is ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for oxazepam in urine. The tests provide only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.

Type of Use (Select one or both, as applicable)

> Prescription Use (Part 21 CFR 801 Subpart D)

> Over-The-Counter Use (21 CFR 801 Subpart C)

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## 510(k) SUMMARY k150162

- February 24, 2015 1. Date: 2. Submitter W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706 3. Contact person:
John Wan W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706 Telephone: [phone] Fax: [phone] Email: [e-mail]

- First Sign® Drug of Abuse Cup Test 4. Device Name: First Sign® Drug of Abuse Dip Card Test
#### Classification: Class II

| Product<br>Code | CFR # | Panel |
|-----------------|----------------------------------------------|------------|
| LAF | 21 CFR, 862.3610 Methamphetamine Test System | Toxicology |
| JXM | 21 CFR, 862.3170 Benzodiazepine Test System | Toxicology |
| DJG | 21 CFR, 862.3650 Opiate Test System | Toxicology |

- 5. Predicate Devices:
K052115 First Check Multi Drug Cup 12

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## 6. Intended Use

First Sign™ Drug of Abuse Tests are immunochromatographic assays for the qualitative determination of Oxazepam , Methamphetamine , and Morphine in human urine at cut-off concentrations of 300 ng/mL, 1000 ng/mL, and 2000 ng/mL, respectively. The tests are available in a Cup format and a Dip Card format.

The tests may yield preliminary positive results even when prescription drug Oxazepam is ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for oxazepam in urine. The tests provide only preliminary test results. A more specific alternative chemical must be used in order to obtain a confirmed analytical result. Gas Chromatography/Mass Spectrometry is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

For in vitro diagnostic use only. The tests are intended for over-the-counter and for prescription use.

- 7. Device Description
First Sign™ Drug of Abuse Tests are immunochromatographic assays. Each assay test is a lateral flow system for the qualitative detection of Oxazepam , Methamphetamine , and Morphine in human urine. The products are single-use in vitro diagnostic devices, which come in the formats of DipCards or Cups. Each test kit contains a Test Device (in one of the two formats), a package insert and a urine cup for sample collection. Each test device is sealed with a desiccant in an aluminum pouch.

- 8. Substantial Equivalence Information
A summary comparison of features of the First Sign™ Drug of Abuse Test and the predicate device is provided in Table 1, Table 2 & Table 3.

| Item | Device | Predicate -<br>K052115 |
|--------------------------|-------------------------------------------------------------------------------------------------------------------------------------|------------------------|
| Indication(s)<br>for Use | For the qualitative determination of<br>Oxazepam in human urine. | Same |
| Calibrator | Oxazepam | Same |
| Methodology | Competitive binding, lateral flow<br>immunochromatographic assays based on<br>the principle of antigen antibody<br>immunochemistry. | Same |
| Specimen Type | Human Urine | Same |
| Cut-Off Values | 300 ng/mL | Same |

### Table 1: Features Comparison of First Sign™ Oxazepam Test and the Predicate Device

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| Item | Device | Predicate -<br>K052115 |
|------------------------|------------------------------------------------|---------------------------------|
| Intended<br>Population | For over-the-counter and prescription<br>uses. | For<br>over-the-counter<br>use. |
| Configurations | Cup, Dip Card | Cup |

# Table 2: Features Comparison of First Sign™ Methamphetamine Test and the Predicate Device

| Item | Device | Predicate - K052115 |
|-----------------------|----------------------------------------------------------------------------------------------------------------------------|---------------------------|
| Indication(s) for Use | For the qualitative determination of methamphetamine in human urine. | Same |
| Calibrator | D-methamphetamine | Same |
| Methodology | Competitive binding, lateral flow immunochromatographic assays based on the principle of antigen antibody immunochemistry. | Same |
| Specimen Type | Human Urine | Same |
| Cut-Off Values | 1000 ng/mL | Same |
| Intended Population | For over-the-counter and prescription uses. | For over-the-counter use. |
| Configurations | Cup, Dip Card | Cup |

## Table 3: Features Comparison of First Sign™ Morphine Test and the Predicate Device

| Item | Device | Predicate - K052115 |
|--------------------------|-----------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Indication(s)<br>for Use | For the qualitative determination of<br>morphine in human urine. | For the qualitative<br>determination of<br>opiates in human<br>urine, including<br>morphine. |
| Calibrator | Morphine | Same |
| Methodology | Competitive binding, lateral flow<br>immunochromatographic assays based<br>on the principle of antigen antibody | Same |

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| Item | Device | Predicate - K052115 |
|----------------|---------------------------------------|----------------------|
| | immunochemistry. | |
| Specimen Type | Human Urine | Same |
| Cut-Off Values | 2000 ng/mL | Same |
| Intended | For over-the-counter and prescription | For over-the-counter |
| Population | uses. | use. |
| Configurations | Cup, Dip Card | Cup |

#### 9. Test Principle

First Sign™ Drug of Abuse Tests are rapid tests for the qualitative detection of Oxazepam . Methamphetamine , and Morphine in urine samples. Each assay test is a lateral flow chromatographic immunoassay. During testing, a urine specimen migrates upward by capillary action. If target drugs are present in the urine specimen below its cut-off concentration, it will not saturate the binding sites of its specific antibody (monoclonal mouse antibody) coated on the particles. The antibody-coated particles will then be captured by immobilized drug-conjugate and a visible colored line will show up in the test line region. The colored line will not form in the test line region if the target drug level exceeds its cut-off concentration because it will saturate all the binding sites of the antibody coated on the particles. A band should form in the control region of the devices regardless of the presence of drug or metabolite in the sample.

#### 10. Performance Characteristics

#### 1. Analytical Performance

- a. Precision
Precision studies were carried out for samples with concentrations of -100% cut-off, -50% cut-off, -25% cut-off, at the cut-off, +25% cut-off, +75% cut-off, +75% cut-off and +100% cut-off. These samples were prepared by spiking drug in negative samples. Each drug concentration was confirmed by GC/MS. All sample aliquots were blind-labeled and randomized by the person who prepared samples and did not take part in the sample testing. For each concentration, tests were performed two runs per day for 25 days by three different operators for each format of devices. Different set of operators tested each format. The results obtained are summarized in the following tables:

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## Oxazepam Dip Card Format

| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|------|---------------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | Lot: D4010558 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Drug | Lot: D4010560 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 4-/46+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Drug | Lot: D4010562 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

## Oxazepam Cup Format

| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|---------------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | | | | | | | | | | |
| Lot: D4010559 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010561 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010563 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 4-/46+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

## Methamphetamine (mAMP) Dip Card Format

| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|------|---------------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | Lot: D4010558 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 4-/46+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| | Lot: D4010560 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| | Lot: D4010562 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

## (mAMP) Cup Format

| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|---------------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | | | | | | | | | | |
| Lot: D4010559 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010561 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010563 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

## Morphine Dip Card Format

| | Result | -100%<br>Cut-off | -75%<br>Cut-off | -50%<br>Cut-off | -25%<br>Cut-off | Cut-off | +25%<br>Cut-off | +50%<br>Cut-off | +75%<br>Cut-off | +100%<br>Cut-off |
|---------------|--------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| Drug | | | | | | | | | | |
| Lot: D4010558 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010560 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010562 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 4-/46+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

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| | Result | -100% Cut-off | -75% Cut-off | -50% Cut-off | -25% Cut-off | Cut-off | +25% Cut-off | +50% Cut-off | +75% Cut-off | +100% Cut-off |
|---------------|--------|---------------|--------------|--------------|--------------|---------|--------------|--------------|--------------|---------------|
| Drug | | | | | | | | | | |
| Lot: D4010559 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 2-/48+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010561 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| Lot: D4010563 | | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 3-/47+ | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

#### Morphine Cup Format

#### b. Linearity

Not applicable.

### c. Stability

The devices are stable at 4-30°C (39-86°F) for 24 months based on the accelerated stability study at 50°C. Control materials are not provided with the device. The labeling provides information on how to obtain control materials.

### d. Cut-off

A total of 150 samples equally distributed at concentrations of -50% cut-off; -25% cut-off; cut-off; +25% cut-off; +50% cut-off were tested using three different lots of each device by three different operators. Results were all positive at and above +25% cut-off and all negative at and below -25% cut-off for Oxazepam, Methamphetamine and Morphine. The following cut-off values for the test devices have been verified.

| Test | Calibrator | Cut-off (ng/mL) |
|-------------------------------|-------------------|-----------------|
| One Step Oxazepam Test | Oxazepam | 300 |
| One Step Methamphetamine Test | D-methamphetamine | 1000 |
| One Step Morphine Test | Morphine | 2000 |

### e. Interference

Potential interfering substances found in human urine of physiological conditions were added to drug-free urine and to urine containing target drugs at 25% below and 25% above cut-off levels. These urine samples were tested using three batches of each device for all formats.

Compounds that showed no interference at a concentration of 100µg/mL are summarized in the following tables. There were no differences observed for different formats.

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#### Oxazepam 4-Acetamidophenol Diphenhydramine D.L-Octopamine Acetophenetidin Doxylamine Oxalic acid N-Acetvprocainamide Ecaonine dydrochloride Oxolinic acid Acetvsalicvlic acid Ecqonine methylester Pentobarbital Aminopvrine (-)-Y-Ephedrine Perphenazine Amityptvline Fenoprofen Phencyclidine Amorbarbital Phenelzine Furosemide Amoxicillin Gentisic acid Phenobarbital Ampicillin Hemoglobin Phentermine l-Ascorbic Acid Hydrocortisone L-Phenylephrine O-Hydroxyhippuric acid B-Phenylethylamine D.L-Amphetamine p-Hydroxy-Apormorphine methamphetamine Phenylpropanotamine Aspartame 3-Hydroxytyramine Prednisone Atropine Ibuprofen D.L-Propanolol Benzillic acid Imipramine D-Pseudoephedrine Benzoic acid Iproniazid Quinine Benzoylecaonine (±)Isoproterenol Ranitidine Benzphetamine Isoxsuprine Salicylic acid Bilirubin Ketamine Secobarbital Serotonin (±) Chlorpheniramine Ketoprofen (5-Hydroxytyramine) Caffeine Labetalol Sertraline Cannabidiol Loperamide Sulfamethazine Chloralhvdrate Maprotiline Sulindac Tetrahydrocortisone,(b-D Chloramphenicol Meperidine glucuronide) Chlorothiazide Meprobamate Tetrahydrozoline (±)Chlorpheniramine Thiamine Methadone Methoxyphenamine Thioridazine Chlorpromazine (+) 3,4-Methylenedioxy-Chlorquine amphetamine D.L-Tyrosine (+)3,4-Methylenedioxy-Cholesterol Tolbutamide methamphetamine Clomipramine Nalidixic acid Triamterene Clonidine Nalorphine Trifluoperazine Cocaine hydrochloride Naloxone Trimethoprim Cortisone Naltrexone Triyptamine D.L-Tryptophan (-)cotinine Naproxen

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| Creatinine | Niacinamide | Tyramine |
|-------------------|-------------------|-----------|
| Dextromethlorphan | Nifedipine | Uric acid |
| Diclolrfenac | Norethindrone | Verapamil |
| Diflunisal | D-Norpropoxyphene | Zomepirac |
| Diaoxin | Noscapine | |

# Methamphetamine

| Acetamidophen | Gentisic acid | Oxycodone |
|-----------------------|-----------------------------------|-----------------------------------|
| Acetophenetidin | Glucuronide | Oxymetazoline |
| N-Acetylprocainamide | Glutethimide | Papaverine |
| Acetylsalicylate | Guaifenesin | Penicillin-G |
| Aminopyrine | Hippuric acid | Pentazocine |
| Amitryptyline | Hydralazine | Pentobarbital |
| Amobarbital | Hydrochlorothiazide | Perphenazine |
| Amoxicillin | Hydrocodone | Phencyclidine |
| Ampicillin | Hydrocortisone | Phenelzine |
| Apomorphine | O-Hydroxyhippuric acid | Phenobarbital |
| Aspartame | 3-Hydroxytyramine | Prednisolone |
| Atropine | Ibuprofen | Phenylpropanolamine |
| Benzilic acid | Imipramine | Prednisone |
| Benzoic acid | (-) Isoproterenol | Procaine |
| Benzoylecgonine | Isoxsuprine | Promazine |
| Butabartital | Ketamine | Promethazine |
| Cannabidiol | Ketoprofen | D,L-Propanolol |
| Chloralhydrate | Labetalol | D-Propoxyphene |
| Chloramphenicol | Levorphanol | D-Pseudoephedrine |
| Chlordiazepoxide | Loperamide | Quinidine |
| Chlorothiazide | Loxapine succinate | Quinine |
| Chlorpromazine | Maprotiline | Ranitidine |
| Cholesterol | Meperidine | Salicylic acid |
| Clomipramine | Meprobamate | Secobarbital |
| Clonidine | Methadone | Serotonin (5-<br>Hydroxytyramine) |
| Cocaine hydrochloride | Methaqualone | Sulfamethazine |
| Codeine | Methylphenidal | Sulindac |
| Cortisone | Methyprylon | Temazepam |
| (-) Cotinine | Morphine-3- $\beta$ -Dglucuronide | Tetracycline |
| Creatinine | Nalidixic acid | Tetrahydrocortisone |
| Deoxycorticosterone | Nalorphine | 3 ( $\beta$ -D glucuronide) |
| | | Tetrahydrozoline |

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| Dextromethorphan | Naloxone | Thebaine |
|------------------------|-------------------|-----------------|
| Diazepam | Naltrexone | Thiamine |
| Diclofenac | Naproxen | Thioridazine |
| Diflunisal | Niacinamide | Tolbutamine |
| Digoxin | Nifedipine | Triamterene |
| Diphenhydramine | Norcodein | Trifluoperazine |
| Doxylamine | Norethindrone | Trimethoprim |
| Ecgonine hydrochloride | Noroxymorphone | Trimipramine |
| Ecgonine methyl ester | D-Norpropoxyphene | D, L-Tryptophan |
| Erythromycin | Noscapine | Tyramine |
| β-Estradiol | Nylidrin | D, L-Tyrosine |
| Estrone-3-sulfate | D,L-Octopamine | Uric acid |
| Ethyl-p-aminobenzoate | Oxalic acid | Verapamil |
| Fenoprofen | Oxazepam | Zomepirac |
| Furosemide | Oxolinic acid | |

# Morphine

| 4-Acetamidophenol | Ecgonine methylester | Oxolinic acid |
|----------------------|---------------------------|---------------------|
| Acetophenetidin | (-) -Y -Ephedrine | Oxymetazoline |
| N-Acetylprocainamide | Erythromycin | Papaverine |
| Acetylsalicylic acid | ẞ-Estradiol | Penicillin-G |
| Aminopyrine | Estrone-3-sulfate | Pentazocine |
| Amitryptyline | Ethyl-p-aminobenzoate | Pentobarbital |
| Amobarbital | Fenoprofen | Perphenazine |
| Amoxicillin | Furosemide | Phencyclidine |
| Ampicillin | Gentisic acid | Phenelzine |
| Ascorbic acid | Hemoglobin | Phenobarbital |
| D,L-Amphetamine | Hydralazine | Phentermine |
| Apomorphine | Hydrochlorothiazide | L-Phenylephrine |
| Aspartame | Hydrocortisone | ẞ-Phenylethylamine |
| Atropine | O-Hydroxyhippuric acid | Phenylpropanolamine |
| Benzilic acid | p-Hydroxy methamphetamine | Prednisone |
| Benzoic acid | 3-Hydroxytyramine | D,L-Propanolol |
| Benzoylecgonine | Ibuprofen | D-Propoxyphene |
| Benzphetamine | Imipramine | D-Pseudoephedrine |
| Bilirubin (±) | Iproniazid | Quinidine |
| Brompheniramine | Isoproterenol | Quinine |
| Caffeine | Isoxsuprine | Ranitidine |
| Cannabidiol | Ketamine | Salicylic acid |
| Chloralhydrate | Ketoprofen | Secobarbital |

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

| Chloramphenicol | Labetalol | Serotonin (5-<br>Hydroxytyramine) |
|------------------------|-------------------------------------------|-------------------------------------------|
| Chlordiazepoxide | Loperamide | Sulfamethazine |
| Chlorothiazide | Maprotiline | Sulindac |
| (±) Chlorpheniramine | Meperidine | Temazepam |
| Chlorpromazine | Meprobamate | Tetracycline |
| Chlorquine | Methadone | Tetrahydrocortisone3 (β-D<br>glucuronide) |
| Cholesterol | Methoxyphenamine | Tetrahydrozoline |
| Clomipramine | (+) 3,4-Methylenedioxy-<br>amphetamine | Thiamine |
| Clonidine | (+)3,4-Methylenedioxy-<br>methamphetamine | Thioridazine |
| Cocaine hydrochloride | Nalidixic acid | D, L-Tyrosine |
| Cortisone | Nalorphine | Tolbutamide |
| (-) Cotinine | Naloxone | Triamterene |
| Creatinine | Naltrexone | Trifluoperazine |
| Deoxycorticosterone | Naproxen | Trimethoprim |
| Dextromethorphan | Niacinamide | Trimipramine |
| Diazepam | Nifedipine | Tryptamine |
| Diclofenac | Norethindrone | D, L-Tryptophan |
| Diflunisal | D-Norpropoxyphene | Tyramine |
| Digoxin | Noscapine | Uric acid |
| Diphenhydramine | D,L-Octopamine | Verapamil |
| Doxylamine | Oxalic acid | Zomepirac |
| Ecgonine hydrochloride | Oxazepam | |

- f. Specificity
To test the specificity, drug metabolites and other components that are likely to interfere in urine samples were tested using three batches of each device for all formats. The obtained lowest detectable concentration was used to calculate the cross-reactivity. There were no differences observed for different formats.

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

| Oxazepam | Result | %<br>Cross-Reactivity |
|-------------------------|-------------------------|-----------------------|
| Cut-off=300 ng/mL | | |
| Oxazepam | Positive at 300 ng/mL | 100% |
| Alprazolam | Positive at 125 ng/mL | 240% |
| a-Hydroxyalprazolam | Positive at 2500 ng/mL | 12% |
| Bromazepam | Positive at 1565 ng/mL | 19% |
| Chlordiazepoxide | Positive at 1560 ng/mL | 19% |
| Clobazam | Positive at 65 ng/mL | 462% |
| Clonazepam | Positive at 10000 ng/mL | 3% |
| Clorazepate dipotassium | Positive at 195ng/mL | 154% |
| Delorazepam | Positive at 1560 ng/mL | 19% |
| Desalkylflurazepam | Positive at 1560 ng/mL | 19% |
| Diazepam | Positive at 115 ng/mL | 261% |
| Estazolam | Positive at 165 ng/mL | 182% |
| Flunitrazepam | Positive at 166 ng/mL | 181% |
| Midazolam | Positive at 6500 ng/mL | 5% |
| Nitrazepam | Positive at 300 ng/mL | 100% |
| Norchlordiazepoxide | Positive at 250 ng/mL | 120% |
| Nordiazepam | Positive at 400 ng/mL | 75% |
| Temazepam | Positive at 100 ng/mL | 300% |
| Triazolam | Positive at 2500 ng/mL | 12% |
| D,L-Lorazepam | Negative at ≤ 105 ng/mL | Not Detected |
| Methamphetamine | Negative at ≤ 105 ng/mL | Not Detected |
| Morphine | Negative at ≤ 105 ng/mL | Not Detected |

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

| mAMP<br>(Methamphetamine,<br>Cut-off=1000 ng/mL) | Result | %<br>Cross-Reactivity |
|------------------------------------------------------|-------------------------|-----------------------|
| D-Methamphetamine | Positive at 1000 ng/mL | 100% |
| (+/-)3,4-Methylenedioxy-n-<br>ethylamphetamine(MDEA) | Positive at 41600 ng/mL | 2% |
| D/L-Methamphetamine | Positive at 1000 ng/mL | 100% |
| p-Hydroxymethamphetamine | Positive at 27000 ng/mL | 4% |
| (+/-)3,4-Methylenedioxy<br>methamphetamine (MDMA) | Positive at 8000 ng/mL | 13% |
| L-Methamphetamine | Positive at 10000 ng/mL | 10% |
| Trimethobenzamide | Negative at ≤ 105 ng/mL | Not Detected |
| Chloroquine | Negative at ≤ 105 ng/mL | Not Detected |
| Ephedrine | Negative at ≤ 105 ng/mL | Not Detected |
| Fenfluramine | Negative at ≤ 105 ng/mL | Not Detected |
| Procaine (Novocaine) | Negative at ≤ 105 ng/mL | Not Detected |
| Ranitidine (Zantac) | Negative at ≤ 105 ng/mL | Not Detected |
| D-amphetamine | Negative at ≤ 105 ng/mL | Not Detected |
| L-amphetamine | Negative at ≤ 105 ng/mL | Not Detected |
| Morphine | Negative at ≤ 105 ng/mL | Not Detected |
| Oxazepam | Negative at ≤ 105 ng/mL | Not Detected |

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

| Morphine<br>Cut-off=2000 ng/mL | Result | %<br>Cross-Reactivity |
|--------------------------------|-------------------------|-----------------------|
| Morphine | Positive at 2000 ng/mL | 100% |
| Codeine | Positive at 1000 ng/mL | 200% |
| Ethylmorphine | Positive at 560 ng/mL | 357% |
| Hydrocodone | Positive at 5000 ng/mL | 40% |
| Hydromorphone | Positive at 7315 ng/mL | 27% |
| Levorphanol | Positive at 16000 ng/mL | 13% |
| σ-Monoacetylmorphine | Positive at 1000 ng/mL | 200% |
| Morphine 3-b-D-glucuronide | Positive at 1300 ng/mL | 154% |
| Thebaine | Negative at ≤ 105 ng/mL | Not Detected |
| Norcodeine | Negative at ≤ 105 ng/mL | Not Detected |
| Normorphone | Negative at ≤ 105 ng/mL | Not Detected |
| Oxycodone | Negative at ≤ 105 ng/mL | Not Detected |
| Oxymorphine | Negative at ≤ 105 ng/mL | Not Detected |
| Procaine | Negative at ≤ 105 ng/mL | Not Detected |
| Oxazepam | Negative at ≤ 105 ng/mL | Not Detected |
| Methamphetamine | Negative at ≤ 105 ng/mL | Not Detected |

### g. Effect of Urine Specific Gravity and Urine pH

To investigate the effect of urine specific gravity and urine pH, urine samples with a range of 1.000 to 1.035 specific gravity or urine samples with a range of pH 4 to 9 were spiked with target drugs at 25% below and 25% above cut-off levels. These samples were tested using three batches of each device for all formats. Results were all positive for samples at and above +25% cut-off and all negative for samples at and below -25% Cut-Off. There were no differences observed for different formats.

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

## 2. Comparison Studies

The method comparison studies for the First Sign™ Drug Tests (Cup and Dip Card) for Oxazepam , Methamphetamine , and Morphine were performed in-house with three different laboratory assistants for each format of the device. Operators ran 80 (40 negative and 40 positive) unaltered clinical samples for each drug. The samples were blind labeled and compared to GC/MS results. The results are presented in the tables below:

| | Dip Card format | Negative | Low Negative by GC/MS (less than -50%) | Near Cutoff Negative by GC/MS (Between -50% and cut-off) | Near Cutoff Positive by GC/MS (Between the cut-off and +50%) | High Positive by GC/MS (greater than +50%) |
|----------|-----------------|----------|----------------------------------------|----------------------------------------------------------|--------------------------------------------------------------|--------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 13 | 25 |
| | Negative | 10 | 10 | 20 | 2 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 14 | 25 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 13 | 25 |
| | Negative | 10 | 10 | 20 | 2 | 0 |

## Discordant Results of Oxazepam Dip Card

| Viewer | Sample Number | GC/MS Result | Dipcard Format<br>Viewer Results |
|----------|---------------|--------------|----------------------------------|
| Viewer A | 94638975 | 324 | Negative |
| Viewer A | 83001567 | 325 | Negative |
| Viewer B | 94638975 | 324 | Negative |
| Viewer C | 83002215 | 326 | Negative |
| Viewer C | 94639027 | 328 | Negative |

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

| Cup<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near<br>Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|---------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 14 | 25 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 14 | 25 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 13 | 25 |
| | Negative | 10 | 10 | 20 | 2 | 0 |

## Discordant Results of Oxazepam Cup

| Viewer | Sample Number | GC/MS Result | Cup Format<br>Viewer Results |
|----------|---------------|--------------|------------------------------|
| Viewer A | 83002215 | 326 | Negative |
| Viewer B | 83001567 | 325 | Negative |
| Viewer C | 83001567 | 325 | Negative |
| Viewer C | 94639027 | 328 | Negative |

# Methamphetamine

| Dip Card | | | Low | | Near | |
|----------|----------|----------|---------------------------------------------|-------------------------------------------------------------------------|------------------------------------------------------------------------|--------------------------------------------------------|
| format | | Negative | Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
| Viewer A | Positive | 0 | 0 | 0 | 18 | 21 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 18 | 21 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 17 | 21 |
| | Negative | 10 | 10 | 20 | 2 | 0 |

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

| Viewer | Sample Number | GC/MS Result | DipCard Format Viewer Results |
|----------|---------------|--------------|-------------------------------|
| Viewer A | 83000530 | 1156 | Negative |
| Viewer B | 83001807 | 1094 | Negative |
| Viewer C | 83001811 | 1065 | Negative |
| Viewer C | 83001807 | 1094 | Negative |

Discordant Results of Methamphetamine DipCard

| Cup<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near<br>Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|---------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|--------------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 18 | 21 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 18 | 21 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 17 | 21 |
| | Negative | 10 | 10 | 20 | 2 | 0 |

## Discordant Results of Methamphetamine Cup

| Viewer | Sample Number | GC/MS Result | Cup Format<br>Viewer Results |
|----------|---------------|--------------|------------------------------|
| Viewer A | 83001821 | 1079 | Negative |
| Viewer B | 83001807 | 1094 | Negative |
| Viewer C | 83001821 | 1079 | Negative |
| Viewer C | 83001807 | 1094 | Negative |

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

# Morphine

| DipCard<br>format | Morphine | | | | | |
|-------------------|----------|----------------------------------------------|----------------------------------------------------------------------|-----------------------------------------------------------------------------|-----------------------------------------------------|----|
| | Negative | Low Negative<br>by GC/MS<br>(less than -50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between -50% and<br>cut-off) | Near Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High Positive by<br>GC/MS<br>(greater<br>than +50%) | |
| Viewer A | Positive | 0 | 0 | 0 | 15 | 24 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 15 | 24 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 15 | 24 |
| | Negative | 10 | 10 | 20 | 1 | 0 |

## Discordant Results of Morphine DipCard

| Viewer | Sample Number | GC/MS Result | DipCard Format<br>Viewer Results |
|----------|---------------|--------------|----------------------------------|
| Viewer A | 94639012 | 2119 | Negative |
| Viewer B | 94639012 | 2119 | Negative |
| Viewer C | 83000219 | 2325 | Negative |

| Cup<br>format | | Negative | Low<br>Negative<br>by GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cut-off) | Near Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cut-off<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than<br>+50%) |
|---------------|----------|----------|----------------------------------------------------|-------------------------------------------------------------------------|-----------------------------------------------------------------------------|-----------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 0 | 15 | 24 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer B | Positive | 0 | 0 | 0 | 15 | 24 |
| | Negative | 10 | 10 | 20 | 1 | 0 |
| Viewer C | Positive | 0 | 0 | 0 | 15 | 24 |
| | Negative | 10 | 10 | 20 | 1 | 0 |

# Discordant Results of Morphine Cup

| Viewer | Sample Number | GC/MS Result | Cup Format<br>Viewer Results |
|----------|---------------|--------------|------------------------------|
| Viewer A | 83000219 | 2325 | Negative |
| Viewer B | 83000572 | 2227 | Negative |
| Viewer C | 94639012 | 2119 | Negative |

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

Lay-user study

A lay user study was performed at three intended user sites with 280 lay persons testing the Oxazepam devices, 280 lay persons testing the Methamphetamine devices and 280 lay persons testing the Morphine devices. A total of 139 females and 141 males tested the Oxazepam samples, 138 females and 142 males tested Methamphetamine samples, and 137 females and 143 males tested the Morphine samples. They had diverse educational and professional backgrounds and ranged in age from 21 to > 50 years. Urine samples were prepared at the following concentrations; negative, +/-75%, +/-50%, +/-25% of the cutoff by spiking drugs into drug free-pooled urine specimens. The concentrations of the samples were confirmed by GC/MS. Each sample was aliquoted into individual containers and blind-labeled. Each participant was provided with the package insert, 1 blind labeled sample and a device. The results are summarized below.

| | Number<br>of<br>samples | Oxazepam Concentration<br>by GC/MS<br>(ng/mL) | Lay person results | | The<br>percentage of<br>correct results<br>(%) |
|--------------|-------------------------|-----------------------------------------------|--------------------|--------------------|------------------------------------------------|
| % of Cutoff | | | No. of<br>Positive | No. of<br>Negative | |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 76 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 145 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 222 | 0 | 20 | 100% |
| +25% Cutoff | 20 | 384 | 18 | 2 | 90% |
| +50% Cutoff | 20 | 468 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 542 | 20 | 0 | 100% |

#### Comparison between GC/MS and Lay Person Results (Oxazepam DipCard)

#### Comparison between GC/MS and Lay Person Results (Oxazepam Cup)

| % of Cutoff | Number<br>of<br>samples | Oxazepam Concentration<br>by GC/MS<br>(ng/mL) | Lay person results | | The<br>percentage of<br>correct results<br>(%) |
|--------------|-------------------------|-----------------------------------------------|--------------------|--------------------|------------------------------------------------|
| | | | No. of<br>Positive | No. of<br>Negative | |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 76 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 145 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 222 | 2 | 18 | 90% |
| +25% Cutoff | 20 | 384 | 20 | 0 | 100% |
| +50% Cutoff | 20 | 468 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 542 | 20 | 0 | 100% |

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

| | Number<br>of<br>samples | Methamphetamine<br>Concentration by GC/MS<br>(ng/mL) | Lay person results | | The<br>percentage of<br>correct results<br>(%) |
|--------------|-------------------------|------------------------------------------------------|--------------------|--------------------|------------------------------------------------|
| % of Cutoff | | | No. of<br>Positive | No. of<br>Negative | |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 245 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 488 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 729 | 0 | 20 | 100% |
| +25% Cutoff | 20 | 1212 | 19 | 1 | 95% |
| +50% Cutoff | 20 | 1441 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 1666 | 20 | 0 | 100% |

Comparison between GC/MS and Lay Person Results (Methamphetamine DipCard)

## Comparison between GC/MS and Lay Person Results (Methamphetamine Cup)

| % of Cutoff | Number<br>of<br>samples | Methamphetamine<br>Concentration by GC/MS<br>(ng/mL) | Lay person results | | The<br>percentage of<br>correct results<br>(%) |
|--------------|-------------------------|------------------------------------------------------|--------------------|--------------------|------------------------------------------------|
| | | | No. of<br>Positive | No. of<br>Negative | |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 245 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 488 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 729 | 0 | 20 | 100% |
| +25% Cutoff | 20 | 1212 | 19 | 1 | 95% |
| +50% Cutoff | 20 | 1441 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 1666 | 20 | 0 | 100% |

### Comparison between GC/MS and Lay Person Results (Morphine DipCard)

| % of Cutoff | Number<br>of<br>samples | Morphine Concentration<br>by GC/MS<br>(ng/mL) | Lay person results | | The<br>percentage of<br>correct results<br>(%) |
|--------------|-------------------------|-----------------------------------------------|--------------------|----|------------------------------------------------|
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 527 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 1053 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 1573 | 1 | 19 | 95% |
| +25% Cutoff | 20 | 2652 | 20 | 0 | 100% |
| +50% Cutoff | 20 | 3254 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 3711 | 20 | 0 | 100% |

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

| % of Cutoff | Number<br>of<br>samples | Morphine Concentration<br>by GC/MS<br>(ng/mL) | Lay person results | | The |
|--------------|-------------------------|-----------------------------------------------|--------------------|--------------------|-----------------------------------------|
| | | | No. of<br>Positive | No. of<br>Negative | percentage of<br>correct results<br>(%) |
| -100% Cutoff | 20 | 0 | 0 | 20 | 100% |
| -75% Cutoff | 20 | 527 | 0 | 20 | 100% |
| -50% Cutoff | 20 | 1053 | 0 | 20 | 100% |
| -25% Cutoff | 20 | 1573 | 0 | 20 | 100% |
| +25% Cutoff | 20 | 2652 | 19 | 1 | 95% |
| +50% Cutoff | 20 | 3254 | 20 | 0 | 100% |
| +75% Cutoff | 20 | 3711 | 20 | 0 | 100% |

Comparison between GC/MS and Lay Person Results (Morphine Cup)

Lay-users were also given surveys on the ease of understanding the package insert instructions. All lay users indicated that the device instructions can be easily followed. A Flesch-Kincaid reading analysis was performed on each package insert and the scores revealed a reading Grade Level of 7.

- 3. Clinical Studies
Not applicable.

## 11. Conclusion

Based on the test principle and acceptable performance characteristics including precision, cut-off, interference, specificity and method comparison of the devices, it's concluded that the First Sign™ Drug of Abuse Dip Card Test and First Sign™ Drug of Abuse Cup Test are substantially equivalent to the predicate.