First Sign® Drug of Abuse Buprenorphine Cup Test
First Sign® Drug of Abuse Buprenorphine Dip Card Test
First Sign™ Drug of Abuse Buprenorphine Tests are immunochromatographic assays for the qualitative determination of Buprenorphine, in human urine at cut-off concentration of 10 ng/mL. The tests are available in a Cup format and a Din Card format.

The tests may yield preliminary positive results even when prescription drug Buprenorphine is ingescribed doses; it is not intended to distinguish between prescription use or abuse of this drug. There is no uniformly recognized cutoff concentration level for Buprenorphine 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.

First Sign® Drug of Abuse Butalbital Cup Test
First Sign® Drug of Abuse Butalbital Dip Card Test
First Sign™ Drug of Abuse Butalbital Tests are immunochromatographic assays for the qualitative determination of Butalbital in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.

The tests may yield preliminary positive results even when prescription drug Butalbital 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 Butalbital 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.

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

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.

Device Description

First Sign™ Drug of Abuse Buprenorphine Test, First Sign™ Drug of Abuse Butalbital Test and First Sign™ Drug of Abuse Morphine Test are immunochromatographic assays. Each assay test is a lateral flow system for the qualitative detection of Buprenorphine, or Butalbital or Morphine in human urine. The products are single-use in vitro diagnostic devices, which come in the formats of Dip Cards 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.

AI/ML Overview

Here's a breakdown of the acceptance criteria and study information for the First Sign® Drug of Abuse Buprenorphine, Butalbital, and Morphine tests, based on the provided text:

Acceptance Criteria and Device Performance for Buprenorphine Test

Acceptance Criteria Category	Specific Criteria/Test	Device Performance (Buprenorphine Dip Card)	Device Performance (Buprenorphine Cup)
Precision	Agreement at Cut-Off (10 ng/mL)	3-/47+ (Lot 1, 2, 3)	3-/47+ (Lot 4), 2-/48+ (Lot 5), 3-/47+ (Lot 6)
	Agreement for -100% to -25% Cut-Off (Negative)	50-/0+ for all concentrations	50-/0+ for all concentrations
	Agreement for +25% to +100% Cut-Off (Positive)	50+/0- for all concentrations	50+/0- for all concentrations
Cut-off Verification	Positive at and above +25% cut-off	All positive	All positive
	Negative at and below -25% cut-off	All negative	All negative
Interference	No interference at 100ug/mL for listed compounds	No differences observed for different formats; compounds listed showed no interference	No differences observed for different formats; compounds listed showed no interference
Specificity	Cross-reactivity for related substances	Buprenorphine (100% at 10 ng/mL), Buprenorphine-3-D-Glucuronide (67% at 15 ng/mL), Norbuprenorphine (25% at 40 ng/mL), Norbuprenorphine-3-D-Glucuronide (2% at 500 ng/mL). Morphine, Oxymorphone, Hydromorphone not detected at 100,000 ng/mL.	Same as Dip Card
Effect of Urine Specific Gravity & pH	All positive at and above +25% Cut-Off	All positive	All positive
	All negative at and below -25% Cut-Off	All negative	All negative
Method Comparison (Clinical Samples)	Agreement with GC/MS	(Details in Discordant Results Table)	(Details in Discordant Results Table)
Lay-User Study - Correct Results	-100% to -25% Cut-off (Negative)	100% for all negative concentrations except +25% which was 95% (1 false negative)	100% for all negative concentrations
	+25% to +75% Cut-off (Positive)	95% (1 false negative) for +25% Cutoff, 100% for +50% and +75% Cutoff	95% (1 false negative) for +25% Cutoff, 100% for +50% and +75% Cutoff
Lay-User Study - Ease of Use	Instructions easily followed	All lay users indicated instructions can be easily followed	All lay users indicated instructions can be easily followed

Acceptance Criteria and Device Performance for Butalbital Test

Acceptance Criteria Category	Specific Criteria/Test	Device Performance (Butalbital Dip Card)	Device Performance (Butalbital Cup)
Precision	Agreement at Cut-Off (300 ng/mL)	3-/47+ (Lot 1, 3), 4-/46+ (Lot 2)	3-/47+ (Lot 4, 5, 6)
	Agreement for -100% to -25% Cut-Off (Negative)	50-/0+ for all concentrations	50-/0+ for all concentrations
	Agreement for +25% to +100% Cut-Off (Positive)	50+/0- for all concentrations	50+/0- for all concentrations
Cut-off Verification	Positive at and above +25% cut-off	All positive	All positive
	Negative at and below -25% cut-off	All negative	All negative
Interference	No interference at 100µg/mL for listed compounds	No differences observed for different formats; compounds listed showed no interference	No differences observed for different formats; compounds listed showed no interference
Specificity	Cross-reactivity for related substances	Butalbital (100% at 300 ng/mL), Secobarbital (100% at 300 ng/mL), Amobarbital (10% at 3000 ng/mL), Alphenal (120% at 250 ng/mL), Aprobarbital (150% at 200 ng/mL), Allobarbital (60% at 500 ng/mL), Butabarbital (30% at 1000 ng/mL), Cyclopentobarbital (100% at 300 ng/mL), Pentobarbital (23% at 1300 ng/mL), Phenobarbital (16% at 1900 ng/mL).	Same as Dip Card
Effect of Urine Specific Gravity & pH	All positive at and above +25% Cut-Off	All positive	All positive
	All negative at and below -25% Cut-Off	All negative	All negative
Method Comparison (Clinical Samples)	Agreement with GC/MS	(Details in Discordant Results Table)	(Details in Discordant Results Table)
Lay-User Study - Correct Results	-100% to -25% Cut-off (Negative)	100% for -100% to -50% Cutoff. 95% for -25% Cutoff (1 false positive).	100% for -100% to -50% Cutoff. 95% for -25% Cutoff (1 false positive).
	+25% to +75% Cut-off (Positive)	95% for +25% Cutoff (1 false negative), 100% for +50% and +75% Cutoff.	100% for +25% to +75% Cutoff.
Lay-User Study - Ease of Use	Instructions easily followed	All lay users indicated instructions can be easily followed	All lay users indicated instructions can be easily followed

Acceptance Criteria and Device Performance for Morphine Test

Acceptance Criteria Category	Specific Criteria/Test	Device Performance (Morphine Dip Card)	Device Performance (Morphine Cup)
Precision	Agreement at Cut-Off (300 ng/mL)	2-/48+ (Lot 1, 3), 1-/49+ (Lot 2)	2-/48+ (Lot 4, 6), 3-/47+ (Lot 5)
	Agreement for -100% to -25% Cut-Off (Negative)	50-/0+ for all concentrations	50-/0+ for all concentrations
	Agreement for +25% to +100% Cut-Off (Positive)	50+/0- for all concentrations	50+/0- for all concentrations
Cut-off Verification	Positive at and above +25% cut-off	All positive	All positive
	Negative at and below -25% cut-off	All negative	All negative
Interference	No interference at 100μg/mL for listed compounds	No differences observed for different formats; compounds listed showed no interference	No differences observed for different formats; compounds listed showed no interference
Specificity	Cross-reactivity for related substances	Morphine (100% at 300 ng/mL), 6-Acetylmorphine (40% at 750 ng/mL), Codeine (100% at 300 ng/mL), Morphine-3-B-glucuronide (67% at 450 ng/mL), EthylMorphine (150% at 200 ng/mL), Heroin (43% at 700 ng/mL), Hydromorphone (8% at 4000 ng/mL), Hydrocodone (15% at 2000 ng/mL), Levorphanol (3% at 12000 ng/mL), Thebaine (0.3% at 90000 ng/mL). Oxycodone, Procaine not detected at 100,000 ng/mL.	Same as Dip Card
Effect of Urine Specific Gravity & pH	All positive at and above +25% Cut-Off	All positive	All positive
	All negative at and below -25% Cut-Off	All negative	All negative
Method Comparison (Clinical Samples)	Agreement with GC/MS	(Details in Discordant Results Table)	(Details in Discordant Results Table)
Lay-User Study - Correct Results	-100% to -25% Cut-off (Negative)	100% for -100% to -50% Cutoff. 95% for -25% Cutoff (1 false positive).	100% for -100% to -25% Cutoff.
	+25% to +75% Cut-off (Positive)	100% for +25% to +75% Cutoff.	95% for +25% Cutoff (1 false negative), 100% for +50% and +75% Cutoff.
Lay-User Study - Ease of Use	Instructions easily followed	All lay users indicated instructions can be easily followed	All lay users indicated instructions can be easily followed

Study Details Proving Device Meets Acceptance Criteria:

Sample sizes used for the test set and the data provenance:
- Precision Studies:
  - For each drug (Buprenorphine, Butalbital, Morphine) and each format (Cup, Dip Card), 3 different lots of devices were tested.
  - Each lot was tested with 9 concentrations (e.g., -100% cut-off to +100% cut-off).
  - At each concentration, tests were performed two runs per day for 25 days, by three different operators.
  - This totals to: 3 lots * 9 concentrations * 2 runs/day * 25 days = 1350 tests per operator * 3 operators = 4050 tests per lot. This figure appears to be incorrect considering the "50-/0+" or "3-/47+" reported results which suggest 50 tests per concentration per lot.
  - Given the "50-/0+" format in the tables, it implies 50 tests per concentration per lot. So, for 9 concentrations and 3 lots, this would be 9 * 50 * 3 = 1350 tests for each format (Dip Card/Cup) within the precision study section.
  - Data Provenance: The samples were prepared by "spiking drug in negative urine samples." These were likely laboratory-prepared samples. "Each drug concentration was confirmed by GC/MS."
- Cut-off Verification:
  - A total of 150 samples (equally distributed at -50%, -25%, cut-off, +25%, +50% concentrations) were tested.
  - Performed using three different lots of each device by three different operators.
- Interference & Specificity:
  - Samples were prepared by adding potential interfering substances to drug-free urine and to urine containing target drugs at +/- 25% cut-off levels.
  - These urine samples were tested using three lots of each device for all formats. The number of individual samples is not explicitly stated but implies sufficient testing to cover all listed interfering substances and cross-reactants at specified concentrations.
- Effect of Urine Specific Gravity & pH:
  - Urine samples with varying specific gravity (1.000 to 1.035) and pH (4 to 9) were spiked with target drugs at +/- 25% cut-off levels.
  - These samples were tested using three lots of each device for all formats. The number of individual samples is not explicitly stated.
- Method Comparison Studies (Clinical Samples):
  - 80 unaltered clinical samples (40 negative and 40 positive) were run for each drug (Buprenorphine, Butalbital, Morphine) and each format (Cup, Dip Card).
  - Data Provenance: "unaltered clinical samples." The country of origin is not specified but implicitly US as this is an FDA submission. These are retrospective given they are "clinical samples" compared to a reference method from what is implied a clinical laboratory setting.
- Lay-User Study:
  - 280 lay persons tested the devices for each drug (Buprenorphine, Butalbital, Morphine).
  - Urine samples were prepared at negative, +/-75%, +/-50%, +/-25% of the cutoff by spiking drugs into drug-free pooled urine specimens. 7 concentrations were tested, with 20 samples per concentration.
  - This means 7 concentrations * 20 samples = 140 samples were used per drug/format in the lay-user study.
  - Each participant was provided with 1 blind-labeled sample.
  - Data Provenance: Laboratory-prepared spiked urine samples.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- For the Method Comparison Studies, the ground truth was established by Gas Chromatography/Mass Spectrometry (GC/MS) results. This is an analytical method, not human expert interpretation. Therefore, no human experts were used to establish ground truth for this portion in the traditional sense.
- For Lay-User studies, the "ground truth" for the samples was the known concentration of the spiked drug, confirmed by GC/MS.
Adjudication method (e.g. 2+1, 3+1, none) for the test set:
- Precision, Cut-off, Interference, Specificity, Specific Gravity/pH studies: It's not explicitly stated that an adjudication method was used between readers/operators. For precision, three operators participated, and results were summarized, but no formal adjudication process between them is described.
- Method Comparison Studies: Three different laboratory assistants ran the tests, and their results were compared to GC/MS. No adjudication method between the assistants is explicitly mentioned.
- Lay-User Study: Each lay person tested one blind-labeled sample. No adjudication is applicable here as each person provided a single result for their assigned sample.
If a multi-reader multi-case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance:
- No MRMC comparative effectiveness study was mentioned. The studies focus on the performance of the device itself (standalone) and its interpretation by users (lay-user study), which is a single-reader, single-case scenario in the context of the device's output. There is no AI component mentioned in the document, so no improvement due to AI assistance can be reported.
If a standalone (i.e. algorithm only without human-in-the loop performance) was done:
- The "First Sign® Drug of Abuse" tests are described as "immunochromatographic assays" and "lateral flow systems," which means they are physical test strips/cups that display visual results (lines). The performance studies (precision, specificity, interference, etc.) directly reflect the standalone analytical performance of these devices, as their reactivity with known concentrations and substances is measured. The "Method Comparison Studies" with laboratory assistants and the "Lay-user studies" do involve human interpretation of these visual results, but the core analytical reaction is standalone.
The type of ground truth used (expert consensus, pathology, outcomes data, etc):
- The primary ground truth used across all analytical performance studies (precision, cut-off, interference, specificity, specific gravity/pH) and method comparison studies was Gas Chromatography/Mass Spectrometry (GC/MS) results for drug concentrations in urine samples. This is considered a gold standard analytical method.
- For the lay-user study, the "ground truth" was the known spiked concentration in the urine samples, which itself was confirmed by GC/MS.
The sample size for the training set:
- The document does not describe a computational algorithm or AI model that requires a training set. The devices are immunochromatographic assays. Therefore, the concept of a "training set" in the context of machine learning is not applicable here.
How the ground truth for the training set was established:
- As there is no AI/algorithm and thus no "training set," this question is not applicable.

Summary

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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 is circular, with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter of the circle. In the center of the circle is a stylized image of a caduceus, which is a symbol of medicine.

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

November 9, 2015

W.H.P.M., INC. C/O JOE SHIA REGULATORY CONSULTANT 504E DIAMOND AVE., SUITE I GAITHERSBURG MD 20877

Re: K152551 Trade/Device Name: First Sign® Drug of Abuse Buprenorphine Cup Test First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Butalbital Cup Test First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Morphine Cup Test First Sign® Drug of Abuse Morphine Dip Card Test Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: II Product Code: DJG, DIS Dated: September 1, 2015 Received: September 8, 2015

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

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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.

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 (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/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 (800) 638-2041 or (301) 796-7100 or at its Internet address

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

Sincerely yours.

Courtney H. Lias -S

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) K152551

Device Name

First Sign® Drug of Abuse Buprenorphine Cup Test; First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Butalbital Cup Test; First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Morphine Cup Test; First Sign® Drug of Abuse Morphine Dip Card Test

Indications for Use (Describe)

First Sign® Drug of Abuse Buprenorphine Cup Test

First Sign® Drug of Abuse Buprenorphine Dip Card Test

First Sign™ Drug of Abuse Buprenorphine Tests are immunochromatographic assays for the qualitative determination of Buprenorphine, in human urine at cut-off concentration of 10 ng/mL. The tests are available in a Cup format and a Din Card format.

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

First Sign® Drug of Abuse Butalbital Cup Test

First Sign® Drug of Abuse Butalbital Dip Card Test

First Sign™ Drug of Abuse Butalbital Tests are immunochromatographic assays for the qualitative determination of Butalbital in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.

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

First Sign® Drug of Abuse Morphine Cup Test

First Sign® Drug of Abuse Morphine Dip Card Test

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

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

1. Date: November 6, 2015
1. Submitter W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706
1. Contact person: John Wan W.H.P.M., Inc. 5358 Irwindale Ave. Irwindale, CA 91706 Telephone: 626-443-8480 Fax: 626-443-8065 Email: johnwan@whpm.com
First Sign® Drug of Abuse Buprenorphine Cup Test 4. Device Name: First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Butalbital Cup Test First Sign® Drug of Abuse Butalbital Dip Card Test First Sign® Drug of Abuse Morphine Cup Test First Sign® Drug of Abuse Morphine Dip Card Test
Buprenorphine Urine Test Common Name: Butalbital Urine Test Morphine Urine Test

Product Code	Class	CFR #	Panel
DJG	Class II	21 CFR, 862.3650 Opiate Test System	Toxicology
DIS	Class II	21 CFR, 862.3150 Barbiturate Test System	Toxicology
DJG	Class II	21 CFR, 862.3650 Opiate Test System	Toxicology

This 510(k) summary is for three drug tests namely. BUP, Butalbital and MOP tests. The performance studies including the lay user study were performed on each of the three durg tests (BUP, Butalbital and MOP tests) separately.

1. Predicate Devices:
  K122809, Advin Multi Drug Screen Test K041712 RapidTec 4

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6. Intended Use

First Sign® Drug of Abuse Buprenorphine Cup Test First Sign® Drug of Abuse Buprenorphine Dip Card Test First Sign® Drug of Abuse Buprenorphine Tests are immunochromatographic assays for the qualitative determination of Buprenorphine, in human urine at cut-off concentration of 10 ng/mL. The tests are available in a Cup format and a Dip Card format.

The tests may vield preliminary positive results even when prescription drug Buprenorphine 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 Buprenorphine 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.

First Sign® Drug of Abuse Butalbital Cup Test

First Sign® Drug of Abuse Butalbital Dip Card Test

First Sign® Drug of Abuse Butalbital Tests are immunochromatographic assays for the qualitative determination of Butalbital in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.

The tests may vield preliminary positive results even when prescription drug Butalbital 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 Butalbital 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.

First Sign® Drug of Abuse Morphine Cup Test

First Sign® Drug of Abuse Morphine Dip Card Test

First Sign® Drug of Abuse Morphine Tests are immunochromatographic assays for the qualitative determination of Morphine in human urine at cut-off concentration of 300 ng/mL. The tests are available in a Cup format and a Dip Card format.

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

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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.

1. Device Description
  First Sign™ Drug of Abuse Buprenorphine Test, First Sign™ Drug of Abuse Butalbital Test and First Sign™ Drug of Abuse Morphine Test are immunochromatographic assays. Each assay test is a lateral flow system for the qualitative detection of Buprenorphine, or Butalbital or Morphine in human urine. The products are single-use in vitro diagnostic devices, which come in the formats of Dip Cards 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 candidate device and the predicate device is provided in Table 1, Table 2 & Table 3.

Item	Candidate DeviceFirst Sign® Drug of AbuseBuprenorphine Test	Predicate Device(K122809)Advin Multi DrugScreen Test
Indication(s)for Use	For the qualitative determination ofBuprenorphine in human urine.	Same
Calibrator	Buprenorphine	Same
Methodology	Competitive binding, lateral flowimmunochromatographic assays based onthe principle of antigen antibodyimmunochemistry.	Same
Specimen Type	Human urine	Same
Cut-Off Values	10 ng/mL	Same
IntendedPopulation	For over-the-counter and prescriptionuses.	Forover-the-counteruse.
Configurations	Cup, Dip Card	Cup, Dip Card,Cassette

Table 1: Features Comparison of First Sign® Drug of Abuse Buprenorphine Test and the Predicate Device

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Table 2: Features Comparison of First Sign® Drug of Abuse Butalbital Test and the Predicate Device

	Candidate Device	Predicate Device
Item	First Sign® Drug of Abuse Butalbital Test	(K041712) RapidTec 4
Indication(s) for Use	For the qualitative determination of Butalbital in human urine.	For the qualitative determination of Barbiturates in human urine.
Calibrator	Butalbital	Same
Methodology	Competitive binding, lateral flowimmunochromatographic assays based onthe principle of antigen antibodyimmunochemistry.	Same
Specimen Type	Human urine	Same
Cut-Off Values	300 ng/mL	Same
Intended Population	For over-the-counter and prescription uses.	For prescription use.
Configurations	Cup, Dip Card	Strip

Table 3: Features Comparison of First Sign® Drug of Abuse Morphine Test and the Predicate Device

Item	Candidate DeviceFirst Sign® Drug of Abuse MorphineTest	Predicate Device(K122809)Advin Multi DrugScreen Test
Indication(s)for Use	For the qualitative determination ofMorphine in human urine.	Same
Calibrator	Morphine	Same
Methodology	Competitive binding, lateral flowimmunochromatographic assays based onthe principle of antigen antibodyimmunochemistry.	Same
Specimen Type	Human Urine	Same
Cut-Off Values	300 ng/mL	Same
IntendedPopulation	For over-the-counter and prescriptionuses.	Forover-the-counteruse.

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Item	Candidate DeviceFirst Sign® Drug of Abuse MorphineTest	Predicate Device(K122809)Advin Multi DrugScreen Test
Configurations	Cup, Dip Card	Cup, Dip Card,Cassette

9. Test Principle

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

First Sign® Drug of Abuse Buprenorphine Test

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 urine 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:

	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 1	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-
	Lot 2	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-
	Lot 3	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-

Buprenorphine Dip Card Format

Buprenorphine Cup Format

	Result	-100%Cut-off	-75%Cut-off	-50%Cut-off	-25%Cut-off	+25%Cut-off	+50%Cut-off	+75%Cut-off	+100%Cut-off
Drug	Lot 4	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-

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

Drug	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
Lot 5		50-/0+	50-/0+	50-/0+	50-/0+	2-/48+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 6		50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-

b. Linearity

Not applicable.

c. Stability

The devices are stable at 39-86°F (4-30°C) 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 Buprenorphine. The following cut-off value for the test devices have been verified.

Test	Calibrator	Cut-off (ng/mL)
First Sign® Drug of AbuseBuprenorphine Test	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 at 25% below and 25% above cut-off levels. These urine samples were tested using three lots of each device for all formats.

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

4-Acetamidophenol	β-Estradiol	Oxolinic acid
Acetophenetidin	Estrone-3-sulfate	Oxycodone
N-Acetylprocainamide	Ethyl-p-aminobenzoate	Oxymetazoline
Acetylsalicylic acid	2-ethylidene-1,5-dimethyl-3,3- diphenylpyrrolidine	Papaverine
Aminopyrine	Fenoprofen	Penicillin-G
Amobarbital	Furosemide	Pentazocine hydrochloride
Amoxicillin	Gentisic acid	Pentobarbital
Amphetamine	Hemoglobin	Perphenazine
Ampicillin	Hydralazine	Phencyclidine
L-Ascorbic acid	Hydrochlorothiazide	Phenelzine
Apomorphine	Hydrocodone	Phenobarbital
Aspartame	Hydrocortisone	Phentermine
Atropine	O-Hydroxyhippuric acid	β-Phenylethylamine
Benzilic acid	p-Hydroxyamphetamine	Trans-2-phenylcyclopropylamine hydrochloride
Benzoic acid	p-Hydroxy-methamphetamine	L-Phenylephrine
Benzoylecgonine	3-Hydroxytyramine	Phenylpropanolamine
Benzphetamine	Ibuprofen	Prednisolone
Bilirubin	Iprazid	Prednisone
(±) - Brompheniramine	(±) - Isoproterenol	Procaine
Butalbital	Isoxsuprine	DL-Propranolol
Caffeine	Ketamine	D-Propoxyphene
Cannabidiol	Ketoprofen	D-Pseudoephedrine
Cannabinol	Labetalol	Quinacrine
Chloralhydrate	Loperamide	Quinidine
Chloramphenicol	Methylenedioxyethylamphetamine	Quinine
Chlorothiazide	Meperidine	Ranitidine
(±) Chlorpheniramine	Meprobamate	Salicylic acid
Chlorpromazine	Methadone	Secobarbital
Chlorquine	Methamphetamine	Serotonin
Cholesterol	Methoxyphenamine	Sulfamethazine
Clonidine	(±)-3,4-Methylenedioxyamphetamine hydrochloride	Sulindac
Cocaethylene	Methylenedioxymethamphetamine	Tetracycline
Cocaine hydrochloride	Morphine-3-β-Dglucuronide	Tetrahydrocortisone3-(ß-D-glucuronide)
Codeine	Morphine	Tetrahydrozoline
Cortisone	Morphine sulfate	Thiamine
(-) Cotinine	11-nor-Δ9-THC-9-COOH	Thioridazine
Creatinine	Nalidixic acid	DL-Tyrosine
Deoxycorticosterone	Naloxone	Tolbutamide
Dextromethorphan	Naltrexone	Triamterene

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

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

Diflunisal	Niacinamide	Trimethoprim
Digoxin	Nifedipine	Tryptamine
Diphenhydramine	Norcodeine	DL-Tryptophan
Doxylamine	Norethindrone	Tyramine
Ecgonine hydrochloride	D-Norpropoxyphene	Uric acid
Ecgonine methyl ester	Nortriptyline	Verapamil
Ephedrine	Noscapine	Zomepirac
(L) - Epinephrine	Oxalic acid
Erythromycin	Oxazepam

f. Specificity

To test the specificity, drug metabolites and other components that are likely to interfere in urine samples were tested using three lots 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.

Buprenorphine	Result	%
Cut-off=10 ng/mL		Cross-Reactivity
Buprenorphine	Positive at 10 ng/mL	100%
Buprenorphine -3-D-Glucuronide	Positive at 15 ng/mL	67%
Norbuprenorphine	Positive at 40 ng/mL	25%
Norbuprenorphine-3-D-Glucuronide	Positive at 500 ng/mL	2%
Morphine	Negative at 100000 ng/mL	Not Detected
Oxymorphone	Negative at 100000 ng/mL	Not Detected
Hydromorphone	Negative at 100000 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 lots 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.

h. Comparison Studies

The method comparison studies for the First Sign® Drug of Abuse Buprenorphine Test was 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. The samples were blind labeled and compared to GC/MS results. The results are presented in the tables below:

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

Dip Cardformat		Negative	LowNegativeby GC/MS(less than-50%)	Near CutoffNegative byGC/MS(Between-50% andcut-off)	NearCutoffPositive byGC/MS(Betweenthe cut-offand +50%)	HighPositive byGC/MS(greaterthan +50%)
Viewer A	Positive	0	0	1	13	26
	Negative	10	10	19	1	0
Viewer B	Positive	0	0	1	13	26
	Negative	10	10	19	1	0
Viewer C	Positive	0	0	0	13	26
	Negative	10	10	20	1	0

Discordant Results

Viewer	Sample Number	GC/MS (ng/mL) Result	Dipcard Format Viewer Results
Viewer A	2014122568	9.5	Positive
Viewer A	2014122575	10.5	Negative
Viewer B	2014122505	10.8	Negative
Viewer B	2014122327	9.4	Positive
Viewer C	2014122561	10.7	Negative

Cupformat		Negative	LowNegativeby GC/MS(less than-50%)	Near CutoffNegative byGC/MS(Between-50% andcut-off)	NearCutoffPositive byGC/MS(Betweenthe cut-offand +50%)	HighPositive byGC/MS(greaterthan +50%)
Viewer A	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer B	Positive	0	0	1	13	26
	Negative	10	10	19	1	0
Viewer C	Positive	0	0	1	13	26
	Negative	10	10	19	1	0

Discordant Results

Viewer	Sample Number	GC/MS (ng/mL) Result	Cup Format Viewer Results
Viewer A	2014122575	10.5	Negative
Viewer B	2014122764	9	Positive

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

Viewer	Sample Number	GC/MS (ng/mL)Result	Cup FormatViewer Results
Viewer B	2014122561	10.7	Negative
Viewer C	2014122568	9.5	Positive
Viewer C	2014122505	10.8	Negative

i. Lay-user study

A lay user study was performed at three intended user sites with 280 lay persons testing the Buprenorphine devices. A total of 142 females and 138 males tested the 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; 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.

% of Cutoff	Numberofsamples	BuprenorphineConcentration by GC/MS(ng/mL)	Lay person results		Thepercentage ofcorrect results(%)
			No. ofPositive	No. ofNegative
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	2.6	0	20	100%
-50% Cutoff	20	5.2	0	20	100%
-25% Cutoff	20	7.8	0	20	100%
+25% Cutoff	20	13	19	1	95%
+50% Cutoff	20	15.7	20	0	100%
+75% Cutoff	20	18.3	20	0	100%

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

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

	Number	Buprenorphine	Lay person results		The
% of Cutoff	ofsamples	Concentration by GC/MS(ng/mL)	No. ofNo. ofPositiveNegative		percentage ofcorrect results(%)
-100%Cutoff	20	0	0	20	100%
-75%Cutoff	20	2.6	0	20	100%
-50% Cutoff	20	5.2	0	20	100%
-25% Cutoff	20	7.8	1	19	તે તે જે જ જીરી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય
+25% Cutoff	20	13	19	1	તે તે જે જ જીરી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં પ્રાથમિક શાળા, પંચાયતઘર, આંગણવાડી તેમ જ દૂધની ડેરી જેવી સવલતો પ્રાપ્ય થયેલી છે. આ ગામનાં લોકોનો મુખ્ય
+50% Cutoff	20	15.7	20	0	100%
+75% Cutoff	20	18.3	20	0	100%

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

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.

j. Clinical Studies Not applicable.

First Sign® Drug of Abuse Butalbital Test

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 urine 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:

	-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 1	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 2	50-/0+	50-/0+	50-/0+	50-/0+	4-/46+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 3	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-

Butalbital Dip Card Format

Butalbital Cup Format

	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 4	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-
	Lot 5	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-
	Lot 6	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-

b. Linearity

Not applicable.

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

c. Stability
The devices are stable at 39-86°F (4-30°C) 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 Butalbital. The following cut-off value for the test devices have been verified.

Test	Calibrator	Cut-off (ng/mL)
First Sign® Drug of Abuse ButalbitalTest	Butalbital	300

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 lots 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.

Acetaminophen	Erythromycin	11-nor-Δ9-THC-9-COOH
Acetophenetidin	β-Estradiol	O-Hydroxyhippuric acid
Acetylsalicylic acid	Estrone-3-sulfate	D,L-Octopamine
Aminopyrine	Ethyl-p-aminobenzoate	Oxalic acid
Amitriptyline	Fenoprofen	Oxazepam
Amoxicillin	Furosemide	Oxolinic acid
Amphetamine	Gentisic acid	Oxycodone
Ampicillin	Hemoglobin	Oxymetazoline
Apomorphine	Hydralazine	Papaverine
Ascorbic acid	Hydrochlorothiazide	Penicillin-G
Aspartame	Hydrocodone	Pentazocaine
Atropine	Hydrocortisone	Perphenazine
Benzilic acid	p-Hydroxyamphetamine	Phencyclidine
Benzoic acid	p-Hydroxymethamphetamine	Phenelzine
Benzoylecgonine	3-Hydroxytyramine	β-Phenylethlamine
Bilirubin	Ibuprofen	Phenylpropanolamine
Brompheniramine	Imipramine	Prednisolone
Buprenorphine	(-) Isoproterenol	Prednisone
Caffeine	Isoxsuprine	Procaine
Cannabidiol	Ketamine	Promazine
Cannabinol	Ketoprofen	Promethazine
Chloralhydrate	Labetalol	D,L-Propanolol
Chloramphenicol	Levorphanol	D-Propoxyphene
Chlorothiazide	Loperamide	Quinidine
(±)Chlorpheniramine	L-Phenylephrine	Quinine
Chlorpromazine	Maprotiline	Ranitidine
Chlorquine	Meperidine	Salicylic acid
Cholesterol	Meprobamate	Serotonin
Clomipramine	Morphine-3-β-D glucuronide	Sulfamethazine
Clonidine	Methadone	Sulindac
Cocaine hydrochloride	Methamphetamine	Temazepam
Codeine	(±)-3,4-Methylenedioxy- amphetamine hydrochloride	Tetracycline
Cortisone	Methylenedioxy- methamphetamine	Tetrahydrozoline
(-) Cotinine	Morphine	Thebaine
Creatinine	Morphine Sulfate	Thiamine
Deoxycorticosterone	N-Acetylprocainamide	Thioridazine
Dextromethorphan	Nalidixic acid	Triamterene
Diazepam	Naloxone	Trifluoperazine
Diclofenac	Naltrexone	Trimethoprim
Diflunisal	Naproxen	Trimipramine
Digoxin	Niacinamide	Tryptamine
Diphenhydramine	Nifedipine	D, L-Tyrosine
Doxylamine	Norcodein	Uric acid
Ecgonine hydrochloride	Norethindrone	Verapamil
Ecgonine methylester	D-Norpropoxyphene	Zomepirac
(IR,2S)(-)Ephedrine	Noscapine
2-ethylidene-1,5-dimethyl-3, 3- diphenylpyrrolidine	Nortriptyline

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

f. Specificity

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

Butalbital	Result	% Cross-Reactivity
Cut-off=300 ng/mL)
Butalbital	Positive at 300 ng/mL	100%
Secobarbital	Positive at 300 ng/mL	100%
Amobarbital	Positive at 3000 ng/mL	10%
Alphenal	Positive at 250 ng/mL	120%
Aprobarbital	Positive at 200 ng/mL	150%
Allobarbital	Positive at 500 ng/mL	60%
Butabarbital	Positive at 1000 ng/mL	30%
Butethal	Positive at 500 ng/mL	60%
Cyclopentobarbital	Positive at 300 ng/mL	100%
Pentobarbital	Positive at 1300 ng/mL	23%
Phenobarbital	Positive at 1900 ng/mL	16%

g. Effect of Urine Specific Gravity and Urine pH

h. Comparison Studies
The method comparison studies for the First Sign® Drug of Abuse Butalbital Test was 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. The samples were blind labeled and compared to GC/MS results. The results are presented in the tables below:

Dip Cardformat		Negative	LowNegativeby GC/MS(less than-50%)	Near CutoffNegative byGC/MS(Between-50% andcut-off)	NearCutoffPositive byGC/MS(Betweenthe cut-offand +50%)	HighPositive byGC/MS(greaterthan +50%)
Viewer A	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer B	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer C	Positive	0	0	0	13	26
	Negative	10	10	20	1	0

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

Discordant Results
Viewer			GC/MS (ng/mL) DipCard Format
	Sample Number	Result	Viewer Results
Viewer A	94910807	334	Negative
Viewer B	94910628	352	Negative
Viewer C	94910445	341	Negative

Cupformat		Negative	LowNegativeby GC/MS(less than-50%)	Near CutoffNegative byGC/MS(Between-50% andcut-off)	NearCutoffPositive byGC/MS(Betweenthe cut-offand +50%)	HighPositive byGC/MS(greaterthan +50%)
Viewer A	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer B	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer C	Positive	0	0	1	13	26
	Negative	10	10	19	1	0

Discordant Results

Viewer	Sample Number	GC/MS (ng/mL)Result	Cup FormatViewer Results
Viewer A	94910807	334	Negative
Viewer B	94910095	339	Negative
Viewer C	94910628	352	Negative
Viewer C	94910808	270	Positive

i. Lay-user study

A lay user study was performed at three intended user sites with 280 lay persons testing the Butalbital devices. A total of 141 females and 139 males tested the Butalbital 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.

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

% of Cutoff	Number	Butalbital Concentration	Lay person results	The percentage
-------------	--------	--------------------------	--------------------	----------------

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

	ofsamples	by GC/MS(ng/mL)	No. ofPositive	No. ofNegative	of correctresults(%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	77	0	20	100%
-50% Cutoff	20	156	0	20	100%
-25% Cutoff	20	234	1	19	95%
+25% Cutoff	20	390	19	1	95%
+50% Cutoff	20	468	20	0	100%
+75% Cutoff	20	547	20	0	100%

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

% of Cutoff	Numberofsamples	Butalbital Concentrationby GC/MS(ng/mL)	Lay person results		The percentageof correctresults(%)
			No. ofPositive	No. ofNegative
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	77	0	20	100%
-50% Cutoff	20	156	0	20	100%
-25% Cutoff	20	234	1	19	95%
+25% Cutoff	20	390	20	0	100%
+50% Cutoff	20	468	20	0	100%
+75% Cutoff	20	547	20	0	100%

j. Clinical Studies
Not applicable.

First Sign® Drug of Abuse Morphine Test

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 urine 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

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

each format of devices. Different set of operators tested each format. The results obtained are summarized in the following tables:

Drug	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
Lot 1	50-/0+	50-/0+	50-/0+	50-/0+	2-/48+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 2	50-/0+	50-/0+	50-/0+	50-/0+	1-/49+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 3	50-/0+	50-/0+	50-/0+	50-/0+	2-/48+	50+/0-	50+/0-	50+/0-	50+/0-
Morphine Cup Format
	Result100%	75%	50%	25%		+25%	+50%	+75%	+100%

Morphine Dip Card Format

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 4	50-/0+	50-/0+	50-/0+	50-/0+	2-/48+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 5	50-/0+	50-/0+	50-/0+	50-/0+	3-/47+	50+/0-	50+/0-	50+/0-	50+/0-
Lot 6	50-/0+	50-/0+	50-/0+	50-/0+	2-/48+	50+/0-	50+/0-	50+/0-	50+/0-

b. Linearity

Not applicable.

c. Stability

The devices are stable at 39-86 (4-30°C) 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 Morphine. The following cut-off value for the test devices have been verified.

Test	Calibrator	Cut-off (ng/mL)
First Sign® Drug of Abuse MorphineTest	Morphine	300

e. Interference

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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.

Acebutolol	Ecgonine methylester	p-Hydroxymethamphetamine
Acetopromazine	(-) Y Ephedrine	Papaverine
4-Acetamidophenol	Erythromycin	Penicillin-G
Acetophenetidin	β-Estradiol	Pentazocine
N-Acetylprocainamide	Estrone-3-sulfate	Pentobarbital
Acetylsalicylic acid	Ethyl-p-aminobenzoate	Perphenazine
Aminopyrine	Fenoprofen	Phencyclidine
Amitryptyline	Furosemide	Phenelzine
Amobarbital	Gentisic acid	Phenobarbital
Amoxicillin	Hemoglobin	Phentermine
Ampicillin	Hydralazine	L-Phenylephrine
Ascorbic acid	Hydrochlorothiazide	β-Phenylethlamine
Amphetamine	Hydrocortisone	β-Phenyllethylamine
L-Amphetamine	O-Hydroxyhippuric acid	Phenylpropanolamine
Apomorphine	3-Hydroxytyramine	Prednisolone
Aspartame	Ibuprofen	Prednisone
Atropine	Imipramine	Promazine
Benzilic acid	Iprazid	Promethazine
Benzoic acid	(-) -Isoproterenol	D,L-Propanolol
Benzoylecgonine	Isoxsuprine	D-Propoxyphene
Benzphetamine	Ketamine	D-Pseudoephedrine
Bilirubin	Ketoprofen	Quinidine
Brompheniramine	Labetalol	Quinine
Buprenorphine	Loperamide	Ranitidine
Butalbital	Maprotiline	Salicylic acid
Caffeine	Meprobamate	Secobarbital
Chloralhydrate	Methadone	Serotonin(5-Hydroxytyramine)
Chloramphenicol	Methamphetamine	Sulfamethazine
Chlordiazepoxide	Methoxyphenamine	Sulindac
Chlorothiazide	(+)3,4-Methylenedioxyamphetamine	Temazepam
(±) Chlorpheniramine	Methylenedioxymethamphetamine	Tetracycline
Chlorpromazine	Methylphenidate	Tetrahydrocortisone3(5-Dglucuronide)
Chlorquine	Nalorphine	Tetrahydrozoline

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Cholesterol	Naloxone	Thiamine
Clomipramine	Nalidixic acid	Thioridazine
Clonidine	Naltrexone	D, L-Thyroxine
Cocaine hydrochloride	Naproxen	Tolbutamine
Cortisone	Niacinamide	Triamterene
(-) Cotinine	Nifedipine	Trifluoperazine
Creatinine	Norcodein	Trimethoprim
Deoxycorticosterone	Norethindrone	Trimipramine
Dextromethorphan	D-Norpropoxyphene	Tryptamine
Diazepam	11-nor-Δ9-THC-9-COOH	D, L-Tryptophan
Diclofenac	Noscapine	Tyramine
Diflunisal	Nortriptyline	Tyrosine
Digoxin	D,L-Octopamine	Uric acid
Diphenhydramine	Oxalic acid	Verapamil
Doxylamine	Oxazepam	Zomepirac
2-ethylidene-1,5-dimethyl-3,3- diphenylpyrrolidine	Oxolinic acid
Ecgonine hydrochloride	Oxymetazoline

f. Specificity

Morphine	Result	% Cross-Reactivity
Cut-off=300 ng/mL
Morphine	Positive at 300 ng/mL	100%
6-Acetylmorphine	Positive at 750 ng/mL	40%
Codeine	Positive at 300 ng/mL	100%
Morphine-3- β-glucuronide	Positive at 450 ng/mL	67%
EthylMorphine	Positive at 200 ng/mL	150%
Heroin	Positive at 700 ng/mL	43%
Hydromorphone	Positive at 4000 ng/mL	8%
Hydrocodone	Positive at 2000 ng/mL	15%
Levorphanol	Positive at 12000 ng/mL	3%
Thebaine	Positive at 90000 ng/mL	0.3%
Oxycodone	Negative at 100000 ng/mL	Not Detected
Procaine	Negative at 100000 ng/mL	Not Detected

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g. Effect of Urine Specific Gravity and Urine pH

h. Comparison Studies

The method comparison studies for the First Sign® Drug of Abuse Morphine Test was 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. The samples were blind labeled and compared to GC/MS results. The results are presented in the tables below:

DipCardformat		Negative	LowNegativeby GC/MS(less than-50%)	Near CutoffNegative byGC/MS(Between-50% andcut-off)	Near CutoffPositive byGC/MS(Betweenthe cut-offand +50%)	HighPositive byGC/MS(greaterthan+50%)
Viewer A	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer B	Positive	0	0	1	13	26
	Negative	10	10	19	1	0
Viewer C	Positive	0	0	0	13	26
	Negative	10	10	20	1	0

Discordant Results

Viewer	Sample Number	GC/MS (ng/mL) Result	DipCard Format Viewer Results
Viewer A	94911987	338	Negative
Viewer B	94911597	270	Positive
Viewer B	94911984	320	Negative
Viewer C	94911253	340	Negative

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Cup 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	1	13	26
	Negative	10	10	19	1	0
Viewer B	Positive	0	0	0	13	26
	Negative	10	10	20	1	0
Viewer C	Positive	0	0	0	14	26
	Negative	10	10	20	0	0

Discordant Results

Viewer	Sample Number	GC/MS (ng/mL)Result	Cup FormatViewer Results
Viewer A	94911119	268	Positive
Viewer A	94911984	320	Negative
Viewer B	94911976	345	Negative

i. Lay-user study
A lay user study was performed at three intended user sites with 280 lay persons testing the Morphine devices. A total of 143 females and 137 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.

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

% of Cutoff	Number of samples	Morphine Concentration by GC/MS (ng/mL)	Lay person results		The percentage of correct results (%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	74	0	20	100%
-50% Cutoff	20	148	0	20	100%
-25% Cutoff	20	228	1	19	95%
+25% Cutoff	20	379	20	0	100%
+50% Cutoff	20	443	20	0	100%
+75% Cutoff	20	516	20	0	100%

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% of Cutoff	Numberofsamples	Morphine Concentrationby GC/MS(ng/mL)	Lay person results		Thepercentage ofcorrect results(%)
			No. ofPositive	No. ofNegative
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	74	0	20	100%
-50% Cutoff	20	148	0	20	100%
-25% Cutoff	20	228	0	20	100%
+25% Cutoff	20	379	19	1	95%
+50% Cutoff	20	443	20	0	100%
+75% Cutoff	20	516	20	0	100%

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

j. 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 Buprenorphine Test and First Sign® Drug of Abuse Butalbital Test and First Sign® Drug of Abuse Morphine Test are substantially equivalent to the predicate.

Regulation Number and Section

§ 862.3650 Opiate test system.

(a)
Identification. An opiate test system is a device intended to measure any of the addictive narcotic pain-relieving opiate drugs in blood, serum, urine, gastric contents, and saliva. An opiate is any natural or synthetic drug that has morphine-like pharmocological actions. The opiates include drugs such as morphine, morphine glucoronide, heroin, codeine, nalorphine, and meperedine. Measurements obtained by this device are used in the diagnosis and treatment of opiate use or overdose and in monitoring the levels of opiate administration to ensure appropriate therapy.(b)
Classification. Class II (special controls). An opiate 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).