CR3 Keyless Split Sample Cup Nortriptyline-Buprenorphine

K142609 · Guangzhou Wondfo Biotech Co., Ltd. · LFG · Dec 18, 2014 · Clinical Toxicology

Device Facts

• Record ID: K142609
• Device Name: CR3 Keyless Split Sample Cup Nortriptyline-Buprenorphine
• Applicant: Guangzhou Wondfo Biotech Co., Ltd.
• Product Code: LFG · Clinical Toxicology
• Decision Date: Dec 18, 2014
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3910
• Device Class: Class 2

Intended Use

CR3 Keyless Split Sample Cup Nortriptyline–Buprenorphine is a rapid test for the qualitative detection of Nortriptyline (a major metabolite of Tricyclic Antidepressants) and Buprenorphine in human urine at a cutoff concentration of 1000ng/mL and 10ng/mL, respectively. The test is the first step in a two-step process. The second step is to send the sample for laboratory testing if preliminary positive results are obtained. The test is intended for over-the-counter and for prescription use. The test may yield preliminary positive results even when prescription drugs including Tricyclic Antidepressants and Buprenorphine are ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of these drugs. There are no uniformly recognized cutoff concentration levels for Nortriptyline and Buprenorphine in urine. The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS 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.

Device Story

Rapid, single-use, visually read lateral flow immunoassay cup; detects Nortriptyline (1000ng/mL cutoff) and Buprenorphine (10ng/mL cutoff) in human urine. Principle: competitive binding; urine sample migrates via capillary action across membrane pre-coated with drug-protein conjugates and anti-drug antibody-dye conjugates. Negative result: colored line in Test Region (T) (analyte below cutoff). Positive result: no line in Test Region (analyte above cutoff). Control line (C) confirms proper flow. Includes integrated temperature strip (32-38°C) to verify sample validity. Used by lay users (OTC) or clinicians (prescription) to screen for drug presence; preliminary results require laboratory confirmation via GC/MS. Benefits: provides immediate, point-of-care screening for drug presence.

Clinical Evidence

Bench testing only. Precision/reproducibility studies performed over 25 days. Analytical specificity evaluated against numerous compounds; no interference observed. Method comparison against GC/MS using 80 clinical samples showed high agreement. Lay-user study (n=260) confirmed ability of non-professionals to perform test and interpret results; Flesch-Kincaid grade level <7. Temperature strip accuracy validated against thermometer and user study (n=100).

Technological Characteristics

Lateral flow immunochromatographic assay; mouse monoclonal anti-drug antibody-dye conjugates; fixed drug-protein conjugates; anti-mouse IgG polyclonal antibodies. Single-use cup form factor. Visual readout. Integrated temperature strip (32-38°C). No energy source required. Stable at 4-30°C for 18 months.

Indications for Use

Indicated for qualitative detection of Nortriptyline and Buprenorphine in human urine for OTC and prescription use. Not intended to distinguish between prescription use and abuse. Requires confirmatory testing via GC/MS for positive results.

Regulatory Classification

Identification

A tricyclic antidepressant drugs test system is a device intended to measure any of the tricyclic antidepressant drugs in serum. The tricyclic antidepressant drugs include imipramine, desipramine, amitriptyline, nortriptyline, protriptyline, and doxepin. Measurements obtained by this device are used in the diagnosis and treatment of chronic depression to ensure appropriate therapy.

Special Controls

*Classification.* Class II (special controls). A tricyclic antidepressant drugs test system is not exempt if it is intended for any use other than employment or insurance testing or is intended for Federal drug testing programs. The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9, provided the test system is intended for employment and insurance testing and includes a statement in the labeling that the device is intended solely for use in employment and insurance testing, and does not include devices intended for Federal drug testing programs (*e.g.,* programs run by the Substance Abuse and Mental Health Services Administration (SAMHSA), the Department of Transportation (DOT), and the U.S. military).

Predicate Devices

• UCP Multi-Drug Test Key Cups (k132812)

Submission Summary (Full Text)

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Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is an emblem featuring a stylized depiction of three human profiles facing to the right, with flowing lines suggesting movement or connection.

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

December 18, 2014

GUANGZHOU WONDFO BIOTECH CO., LTD. C/O JOE SHIA LSI INTERNATIONAL INC. 504 EAST DIAMOND AVE. SUITE F GAITHERSBURG MD 20878

Re: K142609

Trade/Device Name: CR3 Keyless Split Sample Cup Nortriptyline-Buprenorphine Regulation Number: 21 CFR 862.3910 Regulation Name: Tricyclic antidepressant drugs test system Regulatory Class: II Product Code: LFG. DJG Dated: October 13, 2014 Received: October 16, 2014

Dear Mr. Joe Shia:

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 -S

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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#### DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

## Indications for Use

Form Approved: OMB No. 0910-0120 Expiration Date: January 31, 2017 See PRA Statement below.

510(k) Number (if known) k142609

Device Name

CR3 Keyless Split Sample Cup Nortriptyline - Buprenorphine

#### Indications for Use (Describe)

CR3 Keyless Split Sample Cup Nortriptyline-Buprenorphine is a rapid test for the qualitative detection of Nortriptyline (a major metabolite of Tricyclic Antidepressants) and Buprenorphine in human urine at a cutoff concentration of 1000ng/mL and 10ng/mL, respectively. The test is the first step in a two-step process. The second step is to send the sample for laboratory testing if preliminary positive results are obtained. The test is intended for over-the-counter and for prescription use.

The test may yield preliminary positive results even when prescription drugs including Tricyclic Antidepressants and Buprenorphine are ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of these drugs. There are no uniformly recognized cutoff concentration levels for Nortriptyline in urine. The test provides only preliminary test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. GC/MS is the preferred confirmatory method. Clinical consideration and professional iudgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive.

For in vitro diagnostic use only.

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

> Prescription Use (Part 21 CFR 801 Subpart D)

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

#### CONTINUE ON A SEPARATE PAGE IF NEEDED.

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#### *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."

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## 510(k) SUMMARY

| | 1. Date: | December 10, 2014 | |
|----|-----------------------|-------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------|
| 2. | Submitter: | | Guangzhou Wondfo Biotech Co., Ltd.<br>No.8 Lizhishan Road, Science City, Luogang District,<br>Guangzhou, P.R. China 510663 |
| | 3. Contact person: | Joe Shia<br>Fax: [phone] | LSI International Inc.<br>504 East Diamond Ave., Suite F<br>Gaithersburg, MD 20878<br>Telephone: [phone]<br>Email:shiajl(@yahoo.com |
| 4. | Device Name: | | CR3 Keyless Split Sample Cup Nortriptyline - Buprenorphine |
| | Classification: | Class II<br>Product<br>Code<br>LFG<br>DIG | CFR #<br>21CFR 862.3910<br>21CFR 862.3650 |
| | 5. Predicate Devices: | k132812 | UCP Multi-Drug Test Key Cups |

- 6. Intended Use
CR3 Keyless Split Sample Cup Nortriptyline-Buprenorphine is a rapid test for the qualitative detection of Nortriptyline (a major metabolite of Tricyclic Antidepressants) and Buprenorphine in human urine at a cutoff concentration of 1000ng/mL, respectively. The test is the first step in a two-step process. The second step is to send the sample for laboratory testing if preliminary positive results are obtained. The test is intended for over-the-counter and for prescription use.

The test may yield preliminary positive results even when prescription drugs including Tricyclic Antidepressants and Buprenorphine are ingested, at prescribed doses; it is not intended to distinguish between prescription use or abuse of these drugs. There are no uniformly recognized cutoff concentration levels for Nortriptyline and Buprenorphine in urine. The test provides only preliminary test results. A more specific alternative chemical

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method must be used in order to obtain a confirmed analytical result. GC/MS 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.

- 7. Device Description
The CR3 Keyless Split Sample Cup Nortriptyline-Buprenorphine test uses immunochromatographic assays for nortriptyline and buprenorphine. The test is a lateral flow, one step system for the qualitative detection of nortriptyline and buprenorphine in human urine.

| Item | Device | Predicate |
|--------------------------|-------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------|
| Indication(s)<br>for use | For the qualitative determination of<br>drugs of abuse in human urine | Same |
| Methodology | Competitive binding, lateral flow<br>immunochromatographic assays<br>based on the principle of antigen<br>antibody immunochemistry. | Same |
| Results | Qualitative | Same |
| Specimen<br>Type | Human urine | Same |
| Cut Off Values | Nortriptyline: 1000ng/ml<br>Buprenorphine: 10ng/ml | Same for<br>Nortriptyline and<br>Buprenorphine |
| Configurations | Cup | Cup |
| Conditions for<br>Use | Over-the-Counter & Prescription<br>Use | Same |

- 8. Substantial Equivalence Information
## 9. Test Principle

The CR3 Keyless Split Sample Cup Nortriptyline - Buprenorphine test is a rapid test for the qualitative detection of Nortriptyline and Buprenorphine in urine samples and contains lateral flow chromatographic immunoassays for nortriptyline and buprenorphine. Each assay uses a mouse monoclonal anti-drug antibody-dye conjugate, fixed drug-protein conjugates, and anti-mouse IgG polyclonal antibodies coated on the test membranes. When the absorbent end of the test is immersed into a urine sample, the urine is absorbed into the

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device by capillary action and mixes with the antibody-dye conjugate, flowing across the pre-coated membrane. At analyte concentrations below the target cut-off, antibody-dye conjugates bind to the drug-protein conjugate immobilized in the Test Region (T) of the device. This produces a colored test line that indicates a negative result. When analyte concentration is above the cut-off, analyte molecules bind to the antibody-dye conjugate, preventing the antibody-dye conjugate from binding to the drug-protein conjugate immobilized in the Test Region (T) of the device. No colored band shows in the test region, indicating a potentially positive result. A band should form in the control region (C) of the device 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, -75% cut-off, -50% cut-off, -25% cut-off, at the cut-off, +25% cut-off, +50% cut-off , +75% cut-off and +100% cut-off. For each concentration, tests were performed two runs per day for 25 days. All sample aliquots were masked and randomized. The results obtained are summarized in the following tables:

| Result<br>TCA | -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 |
|---------------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| W11910601CU5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 44+/6- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| W11910602CU5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 43+/7- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| W11910603CU5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 43+/7- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

A. For Nortriptyline (TCA) testing

B. For Buprenorphine (BUP) testing

| Result<br>BUP | -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 |
|---------------|------------------|-----------------|-----------------|-----------------|---------|-----------------|-----------------|-----------------|------------------|
| W11910601CU5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 42+/8- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| W11910602CU5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 43+/7- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |
| W11910603CU5 | 50-/0+ | 50-/0+ | 50-/0+ | 50-/0+ | 43+/7- | 50+/0- | 50+/0- | 50+/0- | 50+/0- |

- b. Linearity

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Not applicable.

- c. Stability
The CR3 Keyless Split Sample Cup Nortriptyline - Buprenorphine is stable at 4-30°C for 18 months as determined by conducting accelerated and real-time stability testing.

Control materials are not provided with the device. The labeling provides information on how to obtain control materials.

- d. Cut-off
A total of 125 nortriptyline samples and 125 buprenorphine samples equally distributed at concentrations of -50%, -25%, at the cut-off, +25%, +50% of their respective cut-offs were labeled by a person who prepared them and would not participate in the sample testing. These samples were tested using three different lots by three different operators. Results were all positive at +25% and +50% cut-off and all negative at -25% and -50% cut-off for both nortriptyline and buprenorphine. The following cut-off values for the test devices have been verified.

| Test | Calibrator | Cut-off<br>(ng/ml) |
|---------------------|---------------|--------------------|
| Nortriptyline (TCA) | nortriptyline | 1000 |
| Buprenorphine (BUP) | buprenorphine | 10 |

- e. Interference
Potential interfering substances found in human urine of physiological or pathological conditions were added to drug-free urine and to urine containing target drugs (nortriptyline or buprenorphine) at 25% below and 25% above the cut-off. These urine samples were tested using three batches of the CR3Keyless Split Sample Cup Nortriptyline - Buprenorphine by three different operators. Compounds that showed no interference at a concentration of 100µg/mL are summarized below:

## Nortriptyline

| 4-Acetamidophenol | Erythromycin | Oxycodone |
|----------------------|-----------------------|---------------|
| Acetophenetidin | β-Estradiol | Oxymetazoline |
| N-Acetylprocainamide | Estrone-3-sulfate | Papaverine |
| Acetylsalicylic acid | Ethyl-p-aminobenzoate | Penicillin-G |

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| Aminopyrine | Fenoprofen | Pentazocine hydrochloride |
|------------------------|----------------------------------------------------|--------------------------------------------------|
| Amobarbital | Furosemide | Pentobarbital |
| Amoxicillin | Gentisic acid | Perphenazine |
| Ampicillin | Hemoglobin | Phencyclidine |
| L-ascorbic acid | Hydralazine | Phenelzine |
| DL-Amphetamine sulfate | Hydrochlorothiazide | Phenobarbital |
| Apomorphine | Hydrocodone | Phentermine |
| Aspartame | Hydrocortisone | β-Phenylethylamine |
| Atropine | O-Hydroxyhippuric acid | Trans-2-phenylcyclopropyl<br>amine hydrochloride |
| Benzilic acid | p-Hydroxyamphetamine | L-Phenylephrine |
| Benzoic acid | p-Hydroxy- methamphetamine | Phenylpropanolamine |
| Benzoylecgonine | 3-Hydroxytyramine | Prednisolone |
| Benzphetamine | Ibuprofen | Prednisone |
| Bilirubin | Iproniazid | Procaine |
| (±) - Brompheniramine | (±) - Isoproterenol | DL-Propanolol |
| Caffeine | Isoxsuprine | D-Propoxyphene |
| Cannabidiol | Ketamine | D-Pseudoephedrine |
| Cannabinol | Ketoprofen | Quinacrine |
| Chloralhydrate | Labetalol | Quinidine |
| Chloramphenicol | Loperamide | Quinine |
| Chlorothiazide | MDE | Ranitidine |
| (±) Chlorpheniramine | Meperidine | Salicylic acid |
| Chlorpromazine | Meprobamate | Secobarbital |
| Chlorquine | Methadone | Serotonin |
| Cholesterol | (L)Methamphetamine | Sulfamethazine |
| Clonidine | Methoxyphenamine | Sulindac |
| Cocaethylene | (±)-3,4-Methylenedioxyamphetamine hydrochloride | Tetracycline |
| Cocaine hydrochloride | (+)3,4-Methylenedioxymethamphetamine hydrochloride | Tetrahydrocortisone,<br>3-acetate |
| Codeine | Morphine-3-β-D-Glucuronide | Tetrahydrocortisone,<br>3-(β-D-glucuronide) |
| Cortisone | Morphine sulfate | Tetrahydrozoline |
| (-) Cotinine | Nalidixic acid | Thiamine |
| Creatinine | Naloxone | Thioridazine |
| Deoxycorticosterone | Naltrexone | DL-Tyrosine |
| Dextromethorphan | Naproxen | Tolbutamide |
| Diclofenac | Niacinamide | Triamterene |
| Diflunisal | Nifedipine | Trifluoperazine |
| Digoxin | Norcodeine | Trimethoprim |

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| Diphenhydramine |
|------------------------|
| Doxylamine |
| Ecgonine hydrochloride |
| Ecgonine methylester |
| Ephedrine |
| (L) - Epinephrine |

#### Buprenorphine

4-Acetamidophenol Acetophenetidin N-Acetylprocainamide Acetylsalicylic acid Aminopyrine Amobarbital Amoxicillin Ampicillin L-ascorbic acid DL-Amphetamine sulfate Apomorphine Aspartame Atropine Benzilic acid Benzoic acid Benzoylecgonine Benzphetamine Bilirubin (±) - Brompheniramine Caffeine Cannabidiol Cannabinol Chloralhydrate Chloramphenicol Chlorothiazide (±) Chlorpheniramine Chlorpromazine Chlorquine Cholesterol

Clonidine

Norethindrone D-Norpropoxyphene Noscapine Oxalic acid Oxazepam Oxolinic acid

Erythromycin ß-Estradiol Estrone-3-sulfate Ethyl-p-aminobenzoate Fenoprofen Furosemide Gentisic acid Hemoglobin Hydralazine Hydrochlorothiazide Hydrocodone Hydrocortisone O-Hydroxyhippuric acid p-Hydroxyamphetamine p-Hydroxy- methamphetamine 3-Hydroxytyramine Ibuprofen Iproniazid (±) - Isoproterenol Isoxsuprine Ketamine Ketoprofen Labetalol Loperamide MDE Meperidine Meprobamate Methadone (L)Methamphetamine Methoxyphenamine

Tryptamine DL-Tryptophan Tyramine Uric acid Verapamil Zomepirac

Oxycodone Oxymetazoline Papaverine Penicillin-G Pentazocine hydrochloride Pentobarbital Perphenazine Phencyclidine Phenelzine Phenobarbital Phentermine β-Phenylethylamine Trans-2-phenylcyclopropyl amine hydrochloride L-Phenylephrine Phenylpropanolamine Prednisolone Prednisone Procaine DL-Propanolol D-Propoxyphene D-Pseudoephedrine Quinacrine Quinidine Quinine Ranitidine Salicylic acid Secobarbital Serotonin Sulfamethazine Sulindac

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| Cocaethylene | (±)-3,4-Methylenedioxyamphetamine hydrochloride | Tetracycline |
|------------------------|----------------------------------------------------|--------------------------------------------|
| Cocaine hydrochloride | (+)3,4-Methylenedioxymethamphetamine hydrochloride | Tetrahydrocortisone,<br>3-acetate |
| Codeine | Morphine-3-β-Dglucuronide | Tetrahydrocortisone<br>3-(β-D-glucuronide) |
| Cortisone | Morphine sulfate | Tetrahydrozoline |
| (-) Cotinine | Nalidixic acid | Thiamine |
| Creatinine | Naloxone | Thioridazine |
| Deoxycorticosterone | Naltrexone | DL-Tyrosine |
| Dextromethorphan | Naproxen | Tolbutamide |
| Diclofenac | Niacinamide | Triamterene |
| Diflunisal | Nifedipine | Trifluoperazine |
| Digoxin | Norcodeine | Trimethoprim |
| Diphenhydramine | Norethindrone | Tryptamine |
| Doxylamine | D-Norpropoxyphene | DL-Tryptophan |
| Ecgonine hydrochloride | Noscapine | Tyramine |
| Ecgonine methylester | Oxalic acid | Uric acid |
| Ephedrine | Oxazepam | Verapamil |
| (L) - Epinephrine | Oxolinic acid | Zomepirac |

- ﻧﮯ Specificity
To test the specificity, drug metabolites and other components that are likely to be present in urine samples were tested. The target drug (Nortriptyline or Buprenorphine), its drug metabolites and the related compounds were studied. These samples were tested using three batches of the CR3Keyless Split Sample Cup Nortriptyline-Buprenorphine by three different operators. The drug metabolites and other components were tested at different concentrations. The obtained lowest detectable concentration was used to calculate the cross-reactivity. Results are shown in the following tables.

| TCA<br>(Nortriptyline,<br>Cut-off=1000 ng/mL) | Result | %<br>Cross-Reactivity |
|-----------------------------------------------|--------------------------|-----------------------|
| Nortriptyline | Positive at 1000 ng/mL | 100% |
| Nordoxepine | Positive at 1,000 ng/mL | 100% |
| Trimipramiine | Positive at 3,000 ng/mL | 33% |
| Amitriptyline | Positive at 1,500 ng/mL | 67% |
| Promazine | Positive at 1,500 ng/mL | 67% |
| Desipramine | Positive at 200 ng/mL | 500% |
| Imipramine | Positive at 400 ng/mL | 250% |
| Clomipramine | Positive at 12,500 ng/mL | 8% |

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| Doxepine | Positive at 2,000 ng/mL | 50% |
|--------------|--------------------------|-----|
| Maprotiline | Positive at 2,000 ng/mL | 50% |
| Promethazine | Positive at 25,000 ng/mL | 4% |

| BUP<br>(Buprenorphine,<br>Cut-off=10 ng/mL) | Result | %<br>Cross-Reactivity |
|---------------------------------------------|-----------------------|-----------------------|
| Buprenorphine | Positive at 10 ng/mL | 100% |
| Buprenorphine -3-D-Glucuronide | Positive at 15 ng/mL | 67% |
| Norbuprenorphine | Positive at 20 ng/mL | 50% |
| Norbuprenorphine -3-D-Glucuronide | Positive at 200 ng/mL | 5% |
| Morphine | >100,000 | <0.1% |
| Oxymorphone | >100,000 | <0.1% |
| Hydromorphone | >100,000 | <0.1% |

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

Twelve urine samples of normal, high, and low specific gravity ranges (1.000 to 1.035) were collected and spiked with either Nortriptyline or Buprenorphine at 25% below and 25% above the corresponding cut-off level. These samples were tested using three batches of the CR3Keyless Split Sample Cup Nortriptyline-Buprenorphine by three different operators.

The pH of an aliquot negative urine pool was adjusted to pH ranges of 4.00 to 9.00 in 1 pH unit increments and spiked with Nortriptyline or Buprenorphine at 25% below and 25% above the corresponding cut-off levels. These samples were tested using three batches of the CR3Keyless Split Sample Cup Nortriptyline-Buprenorphine by three different operators.

The device performance was found not affected by varying specific gravity and pH.

## 2. Comparison Studies

The method comparison for the CR2 Keyless Split Sample Cup Nortriptyline-Buprenorphine was performed in-house with three laboratory assistants. Operators ran 80 (40 negative and 40 positive) unaltered clinical samples. The samples were masked

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and randomized. The obtained test results are compared to GC/MS results. The results are presented in the table below:

## Nortriptyline

| Group<br>Operators | | Negative | Low<br>Negative by<br>GC/MS<br>(less than<br>-50%) | Near Cutoff<br>Negative by<br>GC/MS<br>(Between<br>-50% and<br>cutoff) | Near Cutoff<br>Positive by<br>GC/MS<br>(Between<br>the cutoff<br>and +50%) | High<br>Positive by<br>GC/MS<br>(greater<br>than +50%) |
|--------------------|----------|----------|----------------------------------------------------|------------------------------------------------------------------------|----------------------------------------------------------------------------|--------------------------------------------------------|
| Viewer A | Positive | 0 | 0 | 3 | 9 | 28 |
| | Negative | 10 | 19 | 8 | 3 | 0 |
| Viewer B | Positive | 0 | 0 | 4 | 9 | 28 |
| | Negative | 10 | 19 | 7 | 3 | 0 |
| Viewer C | Positive | 0 | 0 | 3 | 8 | 28 |
| | Negative | 10 | 19 | 8 | 4 | 0 |

Discordant table:

| Viewer | Sample number | GC/MS result | Viewer result |
|----------|---------------|--------------|---------------|
| Viewer A | TCAC1061 | 919 | positive |
| Viewer A | TCAC1062 | 964 | positive |
| Viewer A | TCAC1063 | 944 | positive |
| Viewer A | TCAC1064 | 1082 | negative |
| Viewer A | TCAC1065 | 1012 | negative |
| Viewer A | TCA 1218 | 1245 | negative |
| Viewer B | TCAC1034 | 754 | positive |
| Viewer B | TCAC1061 | 919 | positive |
| Viewer B | TCAC1062 | 964 | positive |
| Viewer B | TCAC1063 | 944 | positive |
| Viewer B | TCAC1064 | 1082 | negative |
| Viewer B | TCAC1065 | 1012 | negative |
| Viewer B | TCAC1093 | 1237 | negative |
| Viewer C | TCAC1061 | 919 | positive |
| Viewer C | TCAC1062 | 964 | positive |
| Viewer C | TCAC1063 | 944 | positive |
| Viewer C | TCAC1064 | 1082 | negative |
| Viewer C | TCAC1065 | 1012 | negative |
| Viewer C | TCAC1093 | 1237 | negative |
| Viewer C | TCA 1218 | 1245 | negative |

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

| Group<br>Operators | | Negative | Low Negative by GC/MS<br>(less than -50%) | Near Cutoff Negative by GC/MS<br>(Between -50% and cutoff) | Near Cutoff Positive by GC/MS<br>(Between the cutoff and +50%) | High Positive by GC/MS<br>(greater than +50%) |
|--------------------|----------|----------|-------------------------------------------|------------------------------------------------------------|----------------------------------------------------------------|-----------------------------------------------|
| Viewer A | Positive | 0 | 0 | 3 | 13 | 23 |
| | Negative | 10 | 11 | 16 | 4 | 0 |
| Viewer B | Positive | 0 | 0 | 4 | 13 | 23 |
| | Negative | 10 | 11 | 15 | 4 | 0 |
| Viewer C | Positive | 0 | 0 | 5 | 14 | 23 |
| | Negative | 10 | 11 | 14 | 3 | 0 |

Discordant table:

| Viewer | Sample number | GC/MS result | viewer results |
|----------|---------------|--------------|----------------|
| Viewer A | BUPC1063 | 9 | positive |
| Viewer A | BUPC1064 | 9 | positive |
| Viewer A | BUP1217 | 9 | positive |
| Viewer A | BUPC1061 | 11 | negative |
| Viewer A | BUPC1062 | 10 | negative |
| Viewer A | BUPC1093 | 12 | negative |
| Viewer A | BUP1224 | 11 | negative |
| Viewer B | BUPC1063 | 9 | positive |
| Viewer B | BUPC1064 | 9 | positive |
| Viewer B | BUP1216 | 8 | positive |
| Viewer B | BUP1217 | 9 | positive |
| Viewer B | BUPC1061 | 11 | negative |
| Viewer B | BUPC1062 | 10 | negative |
| Viewer B | BUPC1091 | 12 | negative |
| Viewer B | BUP1224 | 11 | negative |
| Viewer C | BUPC1033 | 8 | positive |
| Viewer C | BUPC1065 | 9 | positive |
| Viewer C | BUP1213 | 9 | positive |
| Viewer C | BUP1216 | 8 | positive |
| Viewer C | BUP1217 | 9 | positive |
| Viewer C | BUPC1061 | 11 | negative |
| Viewer C | BUPC1091 | 12 | negative |
| Viewer C | BUP1224 | 11 | negative |

Lay-user study

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

A lay user study was performed at three intended user sites with 260 lay persons, of which, 20 tested for drug-free samples, 120 for nortriptyline samples, 120 for buprenorphine 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; -100%, +/-75%, +/-50%, +/-25% of the cut-off by spiking drug(s) into drug free-pooled urine specimens. The concentrations of the samples were confirmed by GC/MS. Each sample was aliquoted into individual containers, blind-labeled and randomized. Each participant was provided with the package insert, 1 blind labeled sample and a device. The results are summarized below:

| Cup format | | Number<br>of<br>samples | OTC user | | %Agreement<br>With<br>GC/MS |
|---------------|---------------|-------------------------|----------|----------|-----------------------------|
| Drug | Concentration | | Negative | Positive | |
| Drug -free | -100% | 20 | 20 | 0 | 100% |
| | -75% | 20 | 20 | 0 | 100% |
| Nortriptyline | -50% | 20 | 20 | 0 | 100% |
| | -25% | 20 | 17 | 3 | 85% |
| | +25% | 20 | 3 | 17 | 85% |
| | +50% | 20 | 0 | 20 | 100% |
| | +75% | 20 | 0 | 20 | 100% |
| Buprenorphine | -75% | 20 | 20 | 0 | 100% |
| | -50% | 20 | 20 | 0 | 100% |
| | -25% | 20 | 18 | 2 | 90% |
| | +25% | 20 | 3 | 17 | 85% |
| | +50% | 20 | 0 | 20 | 100% |
| | +75% | 20 | 0 | 20 | 100% |

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 the package insert and the score revealed a reading grade level of less than 7.

- 3. Clinical Studies
Not applicable

## 11. Conclusion

Based on the test principle and performance characteristics of the device, it's concluded that CR Keyless Split Sample Cup Nortriptyline -Buprenorphine is substantially equivalent to the predicate.