Healgen Secobarbital Test is an immunochromatographic assay for the qualitative determination of Secobarbital in human urine at a Cut-Off concentration of 300 ng/mL. The test is available in a Strip format, a Cassette format, a Dip Card format and a Cup format.

The test may yield preliminary positive results even when the prescription drug Secobarbital 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 Secobarbital 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. It is intended for prescription and for over-the-counter use.

Healgen Buprenorphine Test is an immunochromatographic assay for the qualitative determination of Buprenorplaine in human urine at a Cut-Off concentration of 10 ng/mL. The test is available in a Strip format, a Cassette format, a Dip Card format and a Cup format.

The test may yield preliminary positive results even when the prescription drug Burrenorphine 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 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. It is intended for prescription and for over-the-counter use.

Healgen Methadone Test is an immunochromatographic assay for the qualitative determination of Methadone in human urine at a Cut-Off concentration of 300 ng/mL. The test is available in a Strip format, a Cassette format, a Dip Card format and a Cup format.

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. It is intended for prescription and for over-the-counter use.

Device Description

Healgen Secobarbital Test. Healgen Buprenorphine Test and Healgen Methadone Test are immunochromatographic assays for Secobarbital, Buprenorphine and Methadone. Each assay test is a lateral flow system for the qualitative detection of Secobarbital, Buprenorphine and Methadone (target analyte) in human urine. The products are in vitro diagnostic devices, which come in the form of: Strips, Cassettes, DipCards, or Cups. Each product contains a Test Device (in one of the four formats), and a package insert. Each test device is sealed with a desiccant in an aluminum pouch.

AI/ML Overview

Here's an analysis of the acceptance criteria and study findings for the Healgen Secobarbital, Buprenorphine, and Methadone Tests, structured as requested:

Acceptance Criteria and Device Performance

The provided document describes the analytical and comparison studies performed to demonstrate the substantial equivalence of the Healgen drug tests. The acceptance criteria are implied by the expected performance in these studies, particularly the accuracy relative to GC/MS at and around the cutoff concentrations.

Table of Acceptance Criteria (Implied) and Reported Device Performance

Test Type	Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance (Summary)
Precision	Accuracy at various concentrations around the cutoff	Consistent and correct results (negative below cutoff, positive above) at +/- 50%, +/- 75%, +/- 100% of the cutoff. Mixed results expected only very close to the cutoff (+/- 25%).	For all three drugs and all four formats, tests performed by three different operators over 25 days showed: - 100% accuracy (50-/0+ or 50+/0-) at -100%, -75%, -50%, +50%, +75%, +100% of cutoff. - Mixed results (e.g., 22-/28+, 25-/25+, 20-/30+) within the +/- 25% range of the cutoff, which is expected for qualitative tests at the decision point.
Cut-off Verification	Ability to distinguish between drug concentrations at and around the cut-off	All samples at or above +25% cut-off should be positive; all samples at or below -25% cut-off should be negative.	For all three drugs, across three different device lots, and three different operators: - All results were positive at and above +25% cut-off. - All results were negative at and below -25% cut-off.
Interference	Absence of interference from common urine substances	No interference at 100µg/mL for listed substances.	No differences observed across different formats for a comprehensive list of compounds at 100µg/mL for all three drugs.
Specificity	Cross-reactivity with related compounds and non-reactivity with unrelated compounds.	Acceptable levels of cross-reactivity for structurally similar compounds and no detection for unrelated compounds (where applicable).	- Secobarbital: 100% cross-reactivity with Amobarbital; varying degrees with Alphenol (40%), Aprobarbital (120%), Butabarbital (12%), Butathal (12%), Butalbital (12%), Cyclopentobarbital (60%), Pentobarbital (12%), Phenobarbital (1%). - Buprenorphine: 100% with Buprenorphine -3-D-Glucuronide; 50% with Norbuprenorphine and Norbuprenorphine-3-D-Glucuronide. No detection of Morphine, Oxymorphone, Hydromorphone at 100,000 ng/mL. - Methadone: 6% cross-reactivity with Doxylamine. No detection of EDDP, EMDP, LAAM HCI, Alpha Methadol at 100,000 ng/mL.
Effect of Urine Specific Gravity and pH	Consistent results across physiological ranges of specific gravity and pH.	All samples at or above +25% cut-off should be positive; all samples at or below -25% cut-off should be negative.	For all three drugs, across various specific gravity (1.000-1.035) and pH (4-9) levels: - All results were positive for samples at and above +25% cut-off. - All results were negative for samples at and below -25% cut-off.
Method Comparison (Clinical Samples)	Agreement with GC/MS results for negative, low negative, near cutoff negative/positive, and high positive samples.	High agreement expected, especially for definitive negative and high positive samples. Discordant results are noted and acceptable near the cutoff.	Across all three drugs and all four formats, for each of three viewers: - 100% agreement for negative, low negative, and high positive samples. - Discordant results (some positives read as negative, or vice versa) observed mainly in the "Near Cutoff Negative" and "Near Cutoff Positive" categories, which is expected for qualitative tests near their decision point. Specific discordant sample numbers and GC/MS values (e.g., 302 ng/mL, 303 ng/mL for Secobarbital, 10.2 ng/mL for Buprenorphine with a 10 ng/mL cutoff) are provided, showing the device performs qualitatively as expected around the cutoff.
Lay-User Study	High percentage of correct results by untrained users, ease of understanding instructions.	High accuracy across various concentrations, especially for definitive positive/negative samples. Instructions should be easily understood.	For all three drugs and all four formats, performed by over 1680 lay persons (560 per drug): - 90-100% correct results for samples at -25% Cutoff, +25% Cutoff. - 100% correct results for samples at -100%, -75%, -50% Cutoff, and +50%, +75% Cutoff. - Package insert instructions were easily followed, and Flesch-Kincaid read-out at Grade Level 7.

Study Details

Sample size used for the test set and the data provenance (e.g., country of origin of the data, retrospective or prospective)
- Precision Studies:
  - For each of the three drugs (Secobarbital, Buprenorphine, Methadone) and each of the four device formats (Strip, Cassette, Dip Card, Cup): 9 drug concentrations (including the cutoff, +/-25%, +/-50%, +/-75%, +/-100%) were tested with 50 replicates per concentration per lot.
  - This means (9 concentrations * 50 replicates * 3 lots) = 1350 tests per format.
  - Total precision tests per drug: 1350 tests/format * 4 formats = 5400 tests.
  - Total precision tests for all three drugs: 5400 tests * 3 drugs = 16,200 individual tests.
  - Data Provenance: Not explicitly stated, but the study was "carried out for samples... prepared by spiking drug in negative samples." This suggests a controlled laboratory setting, likely prospective. The country of origin is not mentioned.
- Cut-off Verification Studies:
  - For each drug: 150 samples (equally distributed across -50%, -25%, cut-off, +25%, +50% cut-off) per drug. These were tested using three different lots of each device by three different operators.
  - Total samples per drug: 150.
  - Total cut-off verification tests for all three drugs: 150 * 3 drugs = 450 samples.
  - Data Provenance: Controlled laboratory setting, prepared by spiking drugs into negative samples; likely prospective. Country of origin not mentioned.
- Interference Studies:
  - Test Set Description: Drug-free urine and target drug urine (at 25% above cut-off) spiked with various potential interfering substances at 100 µg/mL.
  - Sample Size: Not explicitly stated as a number of individual samples, but tested "three batches of each device for all formats" against a comprehensive list of compounds.
  - Data Provenance: Controlled laboratory setting; likely prospective. Country of origin not mentioned.
- Specificity Studies (Cross-Reactivity):
  - Test Set Description: Related drug metabolites and other components.
  - Sample Size: Not explicitly stated as a number of individual samples, but tested "three batches of each device for all formats."
  - Data Provenance: Controlled laboratory setting; likely prospective. Country of origin not mentioned.
- Effect of Urine Specific Gravity and pH Studies:
  - Test Set Description: Urine samples with specific gravity of 1.000 to 1.035, or pH 4 to 9, spiked with target drugs at 25% below and 25% above cut-off levels.
  - Sample Size: Not explicitly stated but tested "three batches of each device for all formats."
  - Data Provenance: Controlled laboratory setting; likely prospective. Country of origin not mentioned.
- Method Comparison (Clinical Samples):
  - For each of the three drugs and each of the four device formats: 80 unaltered clinical samples (40 negative and 40 positive).
  - Total clinical samples: 80 samples/format * 4 formats * 3 drugs = 960 samples.
  - Each sample was run by three different laboratory assistants.
  - Data Provenance: "unaltered clinical samples," blind labeled. The origin (country, retrospective/prospective) is not specified.
- Lay-User Study:
  - Sample Size: 560 lay persons for each drug test (Secobarbital, Buprenorphine, Methadone). Total individuals performing the study: 1680.
  - For each drug, urine samples were prepared at 7 different concentrations per drug: negative, +/-25%, +/-50%, +/-75%, +/-100% of the cutoff. Each concentration had 20 samples. (7 concentrations * 20 samples = 140 samples per drug tested by lay users).
  - Data Provenance: Conducted at "three intended user sites." The demographic breakdown (age, gender, educational/professional backgrounds) is provided for the participants. Likely a prospective study. Country of origin not mentioned.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g., radiologist with 10 years of experience)
- The ground truth for all analytical and comparison studies was established by GC/MS (Gas Chromatography/Mass Spectrometry). This is a highly accurate and widely accepted "gold standard" analytical method for drug detection and quantification in urine.
- The document implies that the GC/MS results themselves serve as the ground truth, rather than human experts interpreting the GC/MS. The qualifications of the individuals operating the GC/MS are not specified, but they would typically be trained laboratory technicians or chemists.
Adjudication method (e.g., 2+1, 3+1, none) for the test set
- For the Method Comparison (Clinical Samples), the samples were "blind labeled and compared to GC/MS results." The results from the three "Viewer" (laboratory assistants) were individually recorded and compared to the GC/MS. There is no explicit "adjudication method" described where multiple human readers' opinions are combined to form a consensus before comparison to ground truth. Instead, individual human readings (referred to as "Viewer A, B, C") were directly compared to the GC/MS ground truth.
- For the Lay-User Study, each participant interpreted their own test and results were compared to GC/MS. No adjudication method among lay users is mentioned.
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
- This document describes the performance of immunochromatographic assay devices for drug detection, which are qualitative diagnostic tests, not AI-powered image analysis tools.
- Therefore, an MRMC comparative effectiveness study comparing human readers with and without AI assistance was NOT performed or applicable for this device type. The "Viewers" mentioned in the method comparison are human operators reading a physical test strip/cassette/cup/dip card.
If a standalone (i.e. algorithm only without human-in-the loop performance) was done
- This device is an immunochromatographic assay. It is a physical test that produces a visual result (colored lines) which is then interpreted by a human. There is no "algorithm only" performance component. The device's performance is intrinsically linked to human interpretation of its visual output.
- The closest to "standalone" performance are the analytical studies (precision, cut-off, interference, specificity) where the chemical reaction and visual readout are evaluated, but even then, a human reads the result.
The type of ground truth used (expert consensus, pathology, outcomes data, etc)
- The primary ground truth used for all performance evaluations (precision, cut-off, interference, specificity, method comparison, lay-user study) was GC/MS (Gas Chromatography/Mass Spectrometry). This is a highly accurate analytical measurement of drug concentration.
The sample size for the training set
- This document describes the performance evaluation of a rapid diagnostic test (immunochromatographic assay). These types of tests are typically developed through chemical and biological engineering, not through machine learning models that require "training sets" in the computational sense.
- Therefore, there is no "training set" sample size listed in this context.
How the ground truth for the training set was established
- As there is no "training set" for a machine learning model, this question is not applicable to the described device.

Summary

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

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized graphic of three human profiles facing to the right, stacked on top of each other.

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

April 24, 2015

HEALGEN SCIENTIFIC LLC C/O JOE SHIA REGULATORY CONSULTANT 504 EAST DIAMOND AVE. SUITE F GAITHERSBURG MD 20877

Re: K150791

Trade/Device Name: Healgen Secobarbital Test (Strip, Cassette, Cup, Dip Card), Healgen Buprenorphine Test (Strip, Cassette, Cup, Dip Card), Healgen Methadone Test (Strip, Cassette, Cup, Dip Card) Regulation Number: 21 CFR 862.3150 Regulation Name: Barbiturate test system Regulatory Class: II Product Code: DIS, DJR, DJG Dated: March 15, 2015 Received: March 25, 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 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

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

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,

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

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

DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration

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

Indications for Use

510(k) Number (if known) K150791

Device Name Healgen Secobarbital Test (Strip, Cassette, Cup, Dip Card) Healgen Buprenorphine Test (Strip, Cassette, Cup, Dip Card) Healgen Methadone Test (Strip, Cassette, Cup, Dip Card)

Indications for Use (Describe)

For in vitro diagnostic use only. It is intended for prescription and for over-the-counter 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)

CONTINUE ON A SEPARATE PAGE IF NEEDED.FORM FDA 3881 (8/14)

PSC Publishing Services (301) 443-6740

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

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

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

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

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

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

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

1. Date: April 21, 2015
1. Submitter: HEALGEN SCIENTIFIC LLC 5213 Maple St Bellaire, TX 77401
1. Contact person: Jiangiu Fang HEALGEN SCIENTIFIC LLC 5213 Maple St Bellaire, TX 77401 Telephone: 713-733-8088 Fax: 713-733-8088 Email: bryan@healgen.com
1. Device Name: Healgen Secobarbital Test (Strip, Cassette, Cup, Dip Card) Healgen Buprenorphine Test (Strip, Cassette, Cup, Dip Card) Healgen Methadone Test (Strip, Cassette, Cup, Dip Card)

Classification:

Product Code	CFR #	Panel
DIS	21 CFR, 862.3150Barbiturate Test System	Toxicology
DJR	21 CFR, 862.3620Methadone Test System	Toxicology
DJG	21 CFR, 862.3650Opiate Test System	Toxicology

1. Predicate Devices: K132812 UCP Multi-Drug Test Key Cups
1. Intended Use / Indications for Use

The test may yield preliminary positive results even when the prescription drug Secobarbital 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 Secobarbital in urine. The test provides only preliminary test results. A more specific alternative chemical must be used in order to obtain a confirmed analytical result. GC/MS is the preferred confirmatory method. Clinical consideration and

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

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. It is intended for prescription and for over-the-counter use.

Healgen Buprenorphine Test is an immunochromatographic assay for the qualitative determination of Buprenorphine in human urine at a Cut-Off concentration of 10 ng/mL. The test is available in a Strip format, a Cassette format, a Dip Card format and a Cup format.

The test may yield preliminary positive results even when the 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 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. It is intended for prescription and for over-the-counter use.

1. Device Description
  Healgen Secobarbital Test. Healgen Buprenorphine Test and Healgen Methadone Test are immunochromatographic assays for Secobarbital, Buprenorphine and Methadone. Each assay test is a lateral flow system for the qualitative detection of Secobarbital, Buprenorphine and Methadone (target analyte) in human urine. The products are in vitro diagnostic devices, which come in the form of: Strips, Cassettes, DipCards, or Cups. Each product contains a Test Device (in one of the four formats), and a package insert. Each test device is sealed with a desiccant in an aluminum pouch.

8. Substantial Equivalence Information

A summary comparison of features of the Healgen Secobarbital Test, Healgen Buprenorphine Test and Healgen Methadone Test and the predicate device is provided in Table 1, Table 2 and Table 3.

Table 1: Features Comparison of Healgen Secobarbital Test and the Predicate Device

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

Item	Device	Predicate – K132812
Intended Use	For the qualitative determination of drugsof abuse in human urine.	Same
Drug Analyte	Secobarbital	the class ofbarbituratesincludingsecobarbital
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.	Same
Configurations	Strip, Cassette, Cup, Dip Card	Cup

Table 2: Features Comparison of Healgen Buprenorphine Test and the Predicate Device

Item	Device	Predicate -K132812
Intended Use	For the qualitative determination ofdrugs of abuse in human urine.	Same
Drug Analyte	Buprenorphine	Same
Methodology	Competitive binding, lateral flowimmunochromatographic assays basedon the principle of antigen antibodyimmunochemistry.	Same
Specimen Type	Human Urine	Same
Cut-Off Values	10 ng/mLSame
IntendedPopulation	For over-the-counter and prescriptionuses.	Same
Configurations	Strip, Cassette, Cup, Dip CardCup

Table 3: Features Comparison of Healgen Methadone Test and the Predicate Device

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

Item	Device	Predicate – K132812
Intended Use	For the qualitative determination of drugs of abuse in human urine.	Same
Drug Analyte	Methadone	Same
Methodology	Competitive binding, lateral flow immunochromatographic assays based on the principle of antigen antibody immunochemistry.	Same
Specimen Type	Human Urine	Same
Cut-Off Values	300 ng/mL	Same
Intended Population	For over-the-counter and prescription uses.	Same
Configurations	Strip, Cassette, Cup, Dip Card	Cup

9. Test Principle

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

10. Performance Characteristics

1. Analytical Performance
- a. Precision

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

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

Secobarbital

Strip Format

	Result
Drug	-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: BAR1201001	50-/0+	50-/0+	50-/0+	50-/0+	22-/28+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BAR1201002	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BAR1201003	50-/0+	50-/0+	50-/0+	50-/0+	23-/27+	50+/0-	50+/0-	50+/0-	50+/0-

Cassette Format

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
Lot: BAR1201004	50-/0+	50-/0+	50-/0+	50-/0+	22-/28+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BAR1201005	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BAR1201006	50-/0+	50-/0+	50-/0+	50-/0+	17-/33+	50+/0-	50+/0-	50+/0-	50+/0-

Dip Card Format

	-50% -		+50% +75% +100%
Drug	Cut-off Cut-off Cut-off Cut-off	Cut-off	Cut-off Cut-off Cut-off Cut-off
Lot: BAR1201007		50-/0+ 50-/0+ 50-/0+ 50-/0+ 23-/27+ 50+/0- 50+/0- 50+/0-
Lot: BAR1201008		50-/0+ 50-/0+ 50-/0+ 50-/0+ 21-/29+ 50+/0- 50+/0- 50+/0- 50+/0-
Lot: BAR1201009		50-/0+ 50-/0+ 50-/0+ 50-/0+ 23-/27+ 50+/0- 50+/0- 50+/0- 50+/0-

CUP Format

	Result
Drug	-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: BAR1201010	50-/0+	50-/0+	50-/0+	50-/0+	27-/23+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BAR1201011	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BAR1201012	50-/0+	50-/0+	50-/0+	50-/0+	22-/28+	50+/0-	50+/0-	50+/0-	50+/0-

Buprenorphine

Strip Format

Drug	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
Lot: BUP1204001	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BUP1204002	50-/0+	50-/0+	50-/0+	50-/0+	24-/26+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BUP1204003	50-/0+	50-/0+	50-/0+	50-/0+	27-/23+	50+/0-	50+/0-	50+/0-	50+/0-

Cassette Format

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

	-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: BUP1204004	50-/0+	50-/0+	50-/0+	50-/0+	26-/24+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BUP1204005	50-/0+	50-/0+	50-/0+	50-/0+	24-/26+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BUP1204006	50-/0+	50-/0+	50-/0+	50-/0+	26-/24+	50+/0-	50+/0-	50+/0-	50+/0-

Dip Card Format

	-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: BUP1204007	50-/0+	50-/0+	50-/0+	50-/0+	28-/22+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BUP1204008	50-/0+	50-/0+	50-/0+	50-/0+	22-/28+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: BUP1204009	50-/0+	50-/0+	50-/0+	50-/0+	28-/22+	50+/0-	50+/0-	50+/0-	50+/0-

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: BUP1204010	50-/0+	50-/0+	50-/0+	50-/0+	26-/24+	50+/0-	50+/0-	50+/0-	50+/0-
	Lot: BUP1204011	50-/0+	50-/0+	50-/0+	50-/0+	24-/26+	50+/0-	50+/0-	50+/0-	50+/0-
	Lot: BUP1204012	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-

Methadone

Strip Format

	-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: MTD1202001	50-/0+	50-/0+	50-/0+	50-/0+	20-/30+	50+/0-	50+/0-	50+/0-
Lot: MTD1202002	50-/0+	50-/0+	50-/0+	50-/0+	27-/23+	50+/0-	50+/0-	50+/0-
Lot: MTD1202003	50-/0+	50-/0+	50-/0+	50-/0+	26-/24+	50+/0-	50+/0-	50+/0-

Cassette Format

Drug	-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: MTD1202004	50-/0+	50-/0+	50-/0+	50-/0+	19-/31+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: MTD1202005	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: MTD1202006	50-/0+	50-/0+	50-/0+	50-/0+	24-/26+	50+/0-	50+/0-	50+/0-	50+/0-

Dip Card Format

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: MTD1202007		50-/0+	50-/0+	50-/0+	50-/0+	28-/22+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: MTD1202008		50-/0+	50-/0+	50-/0+	50-/0+	24-/26+	50+/0-	50+/0-	50+/0-	50+/0-

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

Drug	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
Lot: MTD1202009		50-/0+	50-/0+	50-/0+	50-/0+	27-/23+	50+/0-	50+/0-	50+/0-

CUP Format

	-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: MTD1202010	50-/0+	50-/0+	50-/0+	50-/0+	22-/28+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: MTD1202011	50-/0+	50-/0+	50-/0+	50-/0+	25-/25+	50+/0-	50+/0-	50+/0-	50+/0-
Lot: MTD1202012	50-/0+	50-/0+	50-/0+	50-/0+	22-/28+	50+/0-	50+/0-	50+/0-	50+/0-

b. Linearity

Not applicable.

c. Stability
The devices are stable at 4-30°C for 24 months based on the accelerated stability study at 45°C and real time stability determination at both 4 ℃ and 30℃.

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 Secobarbital, Buprenorphine and Methadone. The following cut-off values for the test devices have been verified.

Test	Calibrator	Cut-off(ng/mL)
Secobarbital Test	Secobarbital	300
Buprenorphine Test	Buprenorphine	10
Methadone Test	Methadone	300

e. Interference

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

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

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.

Acetophenetidin	Ethyl-p-aminobenzoate	Phencyclidine
N-Acetylprocainamide	Fenoprofen	Phenelzine
Acetylsalicylic Acid(Aspirin)	Furosemide	Phentermine
Aminopyrine	Gentisic acid	Phenylephrine-L
Amitriptyline	Hemoglobin	Phenylethylamine
Amoxicillin	Hydralazine	Phenylpropanolamine
D-Amphetamine	(+/-)-4-HydroxyamphetamineHCL	Prednisolone Acetate
L-Amphetamine	Hydrochlorothiazide	Prednisone
Amphetamine Sulfate	Hydrocodone	Procaine(Novocaine)
Ampicinine(Ampicillin)	Hydrocortisone	Promazine
Apomorphine	a -Hydroxyhippuric acid	Promethazine
L-Ascorbic Acid	p-Hydroxymethamphetamine	Propoxyphene,d-
Aspartame	Ibuprofen	Propranolol
Atropine	Imipramine	Pseudoephedrine HCL
Benzilic acid	Isoxsuprine	Quinidine
Benzphetamine	Isoproterenol-(+/-)	Quinine
Bezoic Acid	Ketamine	Ranitidine(Zantac)
Bilirubin	Labetalol	Salicylic Acid
Caffeine	Levorphanol	Secobarbital
Chloramphenicol	Loperamide	Serotonin
ChlordiazepoxideHCL	Maprotiline	Sulfamethazine
Chloroquine	Meprobamate	Sulindac
Chlorothiazide	Methadone	Temazepam
Chlorpheniramine	Methoxyphenamine	11-Nor-△9-Tetrahydrocannabinol
Chlorpromazine	(+/-)-Methylenedioxyamphetamine(MDA)	Tetracycline
Cholesterol	Methylphenidate	Tetrahydrozoline
Clomipramine	Nalbuphine	Thiamine
Clonidine hydrochloride	Nalidixic acid	L-Thyroxine
Cortisone	Naloxone hydrochloride	ThioridazineHydrochloride
Cotinine(-)	Naltrexone hydrochloride	Triamterene
Creatinine	Naproxen	TriflupromazineHydrochloride

Secobarbital

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

Deoxyepinephrine	Niacinamide	Trimethoprim
Dextromethorphan	Nifedipine	Trimipramine
Diazepam	Norethindrone	Tryptamine
Diflunisal	Norpropoxyphene	DL-Tryptophan
Digoxin	Noscapine	Tyramine
Doxylamine	Oxazepam	D/L-Tyrosine
Ecgonine methylester	Oxymetazoline	Uric Acid
R(-)-Epinephrine	Papaverine	Verapamil
Erythromycin	Penicillin	Zomepirac
Estrone-3-sulfate	Perphenazine

Buprenorphine

Acetophenetidin	Ethyl-p-aminobenzoate	Phencyclidine
N-Acetylprocainamide	Fenoprofen	Phenelzine
Acetylsalicylic Acid(Aspirin)	Furosemide	Phenobarbital
Aminopyrine	Gentisic acid	Phentermine
Amitriptyline	Hemoglobin	Phenylephrine-L
Amoxicillin	Hydralazine	Phenylethylamine
Amobarbital	(+/-)-4-HydroxyamphetamineHCL	Phenylpropanolamine
D-Amphetamine	Hydrochlorothiazide	Prednisolone Acetate
L-Amphetamine	Hydrocodone	Prednisone
Amphetamine Sulfate	Hydrocortisone	Procaine(Novocaine)
Ampicinine(Ampicillin)	a -Hydroxyhippuric acid	Promazine
Apomorphine	p-Hydroxymethamphetamine	Promethazine
L-Ascorbic Acid	Ibuprofen	Propoxyphene,d-
Aspartame	Imipramine	Propranolol
Atropine	Isoxsuprine	Pseudoephedrine HCL
Benzilic acid	Isoproterenol-(+/-)	Quinidine
Benzphetamine	Ketamine	Quinine
Bezoic Acid	Labetalol	Ranitidine(Zantac)
Bilirubin	Levorphanol	Salicylic Acid
Caffeine	Loperamide	Secobarbital
Chloramphenicol	Maprotiline	Serotonin
Chlordiazepoxide HCL	Meprobamate	Sulfamethazine
Chloroquine	Methadone	Sulindac
Chlorothiazide	Methoxyphenamine	Temazepam
Chlorpheniramine	(+/-)-Methylenedioxyamphetamine (MDA)	11-Nor-△9-Tetrahydrocannabinol

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

Chlorpromazine	Methylphenidate	Tetracycline
Cholesterol	Nalbuphine	Tetrahydrozoline
Clomipramine	Nalidixic acid	Thiamine
Clonidine hydrochloride	Naloxone hydrochloride	L-Thyroxine
Cortisone	Naltrexone hydrochloride	ThioridazineHydrochloride
Cotinine(-)	Naproxen	Triamterene
Creatinine	Niacinamide	Triflupromazine
		Hydrochloride
Deoxyepinephrine	Nifedipine	Trimethoprim
Dextromethorphan	Norethindrone	Trimipramine
Diazepam	Norpropoxyphene	Tryptamine
Diflunisal	Noscapine	DL-Tryptophan
Digoxin	Oxazepam	Tyramine
Doxylamine	Oxymetazoline	D/L-Tyrosine
Ecgonine methylester	Papaverine	Uric Acid
R(-)-Epinephrine	Penicillin	Verapamil
Erythromycin	Pentobarbital	Zomepirac
Estrone-3-sulfate	Perphenazine

Methadone

Methadone
Acetophenetidin	Ethyl-p-aminobenzoate	Phenelzine
N-Acetylprocainamide	Fenoprofen	Phenobarbital
Acetylsalicylic Acid(Aspirin)	Furosemide	Phentermine
Aminopyrine	Gentisic acid	Phenylephrine-L
Amitriptyline	Hemoglobin	Phenylethylamine
Amoxicillin	Hydralazine	Phenylpropanolamine
Amobarbital	(+/-)-4-HydroxyamphetamineHCL	Prednisolone Acetate
D-Amphetamine	Hydrochlorothiazide	Prednisone
L-Amphetamine	Hydrocodone	Procaine(Novocaine)
Amphetamine Sulfate	Hydrocortisone	Promazine
Ampicinine(Ampicillin)	a -Hydroxyhippuric acid	Promethazine
Apomorphine	p-Hydroxymethamphetamine	Propoxyphene,d-
L-Ascorbic Acid	Ibuprofen	Propranolol
Aspartame	Imipramine	Pseudoephedrine HCL
Atropine	Isoxsuprine	Quinidine
Benzilic acid	Isoproterenol-(+/-)	Quinine
Benzphetamine	Ketamine	Ranitidine(Zantac)

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

Bezoic Acid	Labetalol	Salicylic Acid
Bilirubin	Levorphanol	Secobarbital
Caffeine	Loperamide	Serotonin
Chloramphenicol	Maprotiline	Sulfamethazine
Chlordiazepoxide HCL	Meprobamate	Sulindac
Chloroquine	Methoxyphenamine	Temazepam
Chlorothiazide	(+/-)-Methylenedioxyamphetamine(MDA)	11-Nor-△9-Tetrahydrocannabinol
Chlorpheniramine	Methylphenidate	Tetracycline
Chlorpromazine	Nalbuphine	Tetrahydrozoline
Cholesterol	Nalidixic acid	Thiamine
Clomipramine	Naloxone hydrochloride	L-Thyroxine
Clonidine hydrochloride	Naltrexone hydrochloride	ThioridazineHydrochloride
Cortisone	Naproxen	Triamterene
Cotinine(-)	Niacinamide	TriflupromazineHydrochloride
Creatinine	Nifedipine	Trimethoprim
Deoxyepinephrine	Norethindrone	Trimipramine
Dextromethorphan	Norpropoxyphene	Tryptamine
Diazepam	Noscapine	DL-Tryptophan
Diflunisal	Oxazepam	Tyramine
Digoxin	Oxymetazoline	D/L-Tyrosine
Doxylamine	Papaverine	Uric Acid
Ecgonine methylester	Penicillin	Verapamil
R(-)-Epinephrine	Pentobarbital	Zomepirac
Erythromycin	Perphenazine
Estrone-3-sulfate	Phencyclidine

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

Secobarbital(Cut-off=300 ng/mL)	Result	% Cross-Reactivity
Secobarbital	Positive at 300 ng/mL	100%
Amobarbital	Positive at 300 ng/mL	100%

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

Alphenol	Positive at 750 ng/mL	40%
Aprobarbital	Positive at 250 ng/mL	120%
Butabarbital	Positive at 2500 ng/mL	12%
Butathal	Positive at 2500 ng/mL	12%
Butalbital	Positive at 2500 ng/mL	12%
Cyclopentobarbital	Positive at 500 ng/mL	60%
Pentobarbital	Positive at 2500 ng/mL	12%
Phenobarbital	Positive at 25000 ng/mL	1%

Buprenorphine(Cut-off=10 ng/mL)	Result	% Cross-Reactivity
Buprenorphine	Positive at 10 ng/mL	100%
Buprenorphine -3-D-Glucuronide	Positive at 10 ng/mL	100%
Norbuprenorphine	Positive at 20 ng/mL	50%
Norbuprenorphine-3-D-Glucuronide	Positive at 20 ng/mL	50%
Morphine	Negative at 100000	Not detected
Oxymorphone	Negative at 100000	Not detected
Hydromorphone	Negative at 100000	Not detected

Methadone(Cut-off=300 ng/mL)	Result	% Cross-Reactivity
Methadone	Positive at 300 ng/mL	100%
Doxylamine	Positive at 5000 ng/mL	6%
EDDP	Negative at 100000	Not detected
EMDP	Negative at 100000	Not detected
LAAM HCI	Negative at 100000	Not detected
Alpha Methadol	Negative at 100000	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 of 1.000 to 1.035 specific gravity or urine samples with pH 4 to 9 were spiked with target drugs at 25% below and 25% above cut-off levels. These samples were tested using three batches of each device for all formats. Results were all positive for samples at and above +25% cut-off and all negative for samples at and below -25% Cut-Off. There were no differences observed for different formats.

2. Comparison Studies

The method comparison studies for the Secobarbital Test, the Buprenorphine Test and the Methadone Test were performed in-house with three different laboratory assistants for each format

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

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:

Secobarbital
Stripformat		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	14	24
	Negative	10	15	15	2	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	15	24
	Negative	10	15	15	1	0

Discordant Results of Secobarbital Strip

Viewer	Sample Number	GC/MS Result	Strip FormatViewer Results
Viewer A	BAR06	303	Negative
Viewer A	BAR72	302	Negative
Viewer B	BAR06	303	Negative
Viewer B	BAR72	302	Negative
Viewer C	BAR06	303	Negative

Cassetteformat		Negative	LowNegativeby GC/MS(less than-50%)	NearCutoffNegativeby GC/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	24
	Negative	10	15	15	3	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	13	24
	Negative	10	15	15	3	0

Discordant Results of Secobarbital Cassette

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

Viewer	Sample Number	GC/MS Result	Cassette FormatViewer Results
Viewer A	BAR06	303	Negative
Viewer A	BAR16	308	Negative
Viewer A	BAR72	302	Negative
Viewer B	BAR06	303	Negative
Viewer B	BAR72	302	Negative
Viewer C	BAR72	302	Negative
Viewer C	BAR06	303	Negative
Viewer C	BAR16	308	Negative

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	0	13	24
	Negative	10	15	15	3	0
Viewer B	Positive	0	0	0	13	24
	Negative	10	15	15	3	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Discordant Results of Secobarbital Cup

Viewer	Sample Number	GC/MS Result	Cup Format Viewer Results
Viewer A	BAR06	303	Negative
Viewer A	BAR16	308	Negative
Viewer A	BAR72	302	Negative
Viewer B	BAR06	303	Negative
Viewer B	BAR16	308	Negative
Viewer B	BAR72	302	Negative
Viewer C	BAR06	303	Negative
Viewer C	BAR72	302	Negative

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

Dip Cardformat		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	24
	Negative	10	15	15	3	0
Viewer B	Positive	0	0	0	13	24
	Negative	10	15	15	3	0
Viewer C	Positive	0	0	0	13	24
	Negative	10	15	15	3	0

Discordant Results of Secobarbital Dip Card

Viewer	Sample Number	GC/MS Result	Dip Card FormatViewer Results
Viewer A	BAR06	303	Negative
Viewer A	BAR16	308	Negative
Viewer A	BAR72	302	Negative
Viewer B	BAR06	303	Negative
Viewer B	BAR16	308	Negative
Viewer B	BAR72	302	Negative
Viewer C	BAR06	303	Negative
Viewer C	BAR16	308	Negative
Viewer C	BAR72	302	Negative

Buprenorphine

Stripformat		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	24
	Negative	10	15	15	3	0
Viewer B	Positive	0	0	0	13	24
	Negative	10	15	15	3	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Discordant Results of Buprenorphine Strip

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

Viewer	Sample Number	GC/MS Result	Strip FormatViewer Results
Viewer A	BUP23	10.2	Negative
Viewer A	BUP67	10.6	Negative
Viewer A	BUP12	10.4	Negative
Viewer B	BUP23	10.2	Negative
Viewer B	BUP67	10.6	Negative
Viewer B	BUP12	10.4	Negative
Viewer C	BUP23	10.2	Negative
Viewer C	BUP12	10.4	Negative

Cassetteformat		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	14	24
	Negative	10	15	15	2	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Discordant Results of Buprenorphine Cassette

Viewer	Sample Number	GC/MS Result	Cassette Format Viewer Results
Viewer A	BUP23	10.2	Negative
Viewer A	BUP12	10.4	Negative
Viewer B	BUP23	10.2	Negative
Viewer B	BUP12	10.4	Negative
Viewer C	BUP23	10.2	Negative
Viewer C	BUP12	10.4	Negative

Dip Cardformat		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%)
			0	0	13	24
Viewer A	Positive	0

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

	Negative	10	15	15	3	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	13	24
	Negative	10	15	15	3	0

Discordant Results of Buprenorphine Dip Card

Viewer	Sample Number	GC/MS Result	Dip Card FormatViewer Results
Viewer A	BUP23	10.2	Negative
Viewer A	BUP67	10.6	Negative
Viewer A	BUP12	10.4	Negative
Viewer B	BUP23	10.2	Negative
Viewer B	BUP12	10.4	Negative
Viewer C	BUP23	10.2	Negative
Viewer C	BUP67	10.6	Negative
Viewer C	BUP12	10.4	Negative

Cupformat		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	24
	Negative	10	15	15	3	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Discordant Results of Buprenorphine Cup

Viewer	Sample Number	GC/MS Result	Cup FormatViewer Results
Viewer A	BUP23	10.2	Negative
Viewer A	BUP67	10.6	Negative
Viewer A	BUP12	10.4	Negative
Viewer B	BUP23	10.2	Negative
Viewer B	BUP12	10.4	Negative
Viewer C	BUP23	10.2	Negative
Viewer C	BUP12	10.4	Negative

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

Methadone

Stripformat		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	14	24
	Negative	10	15	15	2	0
Viewer B	Positive	0	0	0	13	24
	Negative	10	15	15	3	0
Viewer C	Positive	0	0	0	13	24
	Negative	10	15	15	3	0

Discordant Results of Methadone Strip

Viewer	Sample Number	GC/MS Result	Strip Format Viewer Results
Viewer A	MTD80	302	Negative
Viewer A	MTD67	303	Negative
Viewer B	MTD80	302	Negative
Viewer B	MTD67	303	Negative
Viewer B	MTD59	306	Negative
Viewer C	MTD80	302	Negative
Viewer C	MTD67	303	Negative
Viewer C	MTD59	306	Negative

Cassetteformat		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	14	24
	Negative	10	15	15	2	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Discordant Results of Methadone Cassette

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

Viewer	Sample Number	GC/MS Result	Cassette Format Viewer Results
Viewer A	MTD80	302	Negative
Viewer A	MTD67	303	Negative
Viewer B	MTD80	302	Negative
Viewer B	MTD67	303	Negative
Viewer C	MTD80	302	Negative
Viewer C	MTD67	303	Negative

Dip Cardformat		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	14	24
	Negative	10	15	15	2	0
Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Discordant Results of Methadone Dip Card

Viewer	Sample Number	GC/MS Result	Dip Card FormatViewer Results
Viewer A	MTD80	302	Negative
Viewer A	MTD67	303	Negative
Viewer B	MTD80	302	Negative
Viewer B	MTD67	303	Negative
Viewer C	MTD80	302	Negative
Viewer C	MTD67	303	Negative

Cup format		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	24
	Negative	10	15	15	3	0

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

Viewer B	Positive	0	0	0	14	24
	Negative	10	15	15	2	0
Viewer C	Positive	0	0	0	14	24
	Negative	10	15	15	2	0

Viewer	Sample Number	GC/MS Result	Cup FormatViewer Results
Viewer A	MTD80	302	Negative
Viewer A	MTD67	303	Negative
Viewer A	MTD59	306	Negative
Viewer B	MTD80	302	Negative
Viewer B	MTD67	303	Negative
Viewer C	MTD80	302	Negative
Viewer C	MTD67	303	Negative

Discordant Results of Methadone Cup

Lay-user study

A lay user study was performed at three intended user sites with 560 lay persons testing each of the Secobarbital, the Buprenorphine and the Methadone devices. Total of 1680 individuals performed the study. A total of 190 females and 370 males tested the Secobarbital samples, 217 females and 343 males tested the Buprenorphine samples and 216 females and 344 males tested the Methadone 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	Number ofsamples	SecobarbitalConcentration 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	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

Comparison between GC/MS and Lay Person Results (Secobarbital Strip)

Comparison between GC/MS and Lay Person Results (Secobarbital Cassette)

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

	Numberofsamples	SecobarbitalConcentration by GC/MS(ng/mL)	Lay person results		The
% of Cutoff			No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

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

	Number	Secobarbital	Lay person results		The
% of Cutoff	ofsamples	Concentration by GC/MS(ng/mL)	No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100%Cutoff	20	0	0	20	100%
-75%Cutoff	20	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

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

	Numberofsamples	SecobarbitalConcentration by GC/MS(ng/mL)	Lay person results		Thepercentage ofcorrect results(%)
% of Cutoff			No. ofPositive	No. ofNegative
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

% of Cutoff	Number	Buprenorphine	Lay person results	The
-------------	--------	---------------	--------------------	-----

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

	ofsamples	Concentration by GC/MS(ng/mL)	No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	2.5	0	20	100%
-50% Cutoff	20	5	0	20	100%
-25% Cutoff	20	7.5	1	19	95%
+25% Cutoff	20	12.5	18	2	90%
+50% Cutoff	20	15	20	0	100%
+75% Cutoff	20	17.5	20	0	100%

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

% of Cutoff	Number of samples	Buprenorphine Concentration by GC/MS (ng/mL)	Lay person results		The percentage of correct results (%)
			No. of Positive	No. of Negative
-100%Cutoff	20	0	0	20	100%
-75% Cutoff	20	2.5	0	20	100%
-50% Cutoff	20	5	0	20	100%
-25% Cutoff	20	7.5	1	19	95%
+25% Cutoff	20	12.5	19	1	95%
+50% Cutoff	20	15	20	0	100%
+75% Cutoff	20	17.5	20	0	100%

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

% of Cutoff	Number of samples	Buprenorphine Concentration by GC/MS (ng/mL)	Lay person results		The percentage of correct results (%)
			No. of Positive	No. of Negative
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	2.5	0	20	100%
-50% Cutoff	20	5	0	20	100%
-25% Cutoff	20	7.5	2	18	90%
+25% Cutoff	20	12.5	18	2	90%
+50% Cutoff	20	15	20	0	100%
+75% Cutoff	20	17.5	20	0	100%

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

% of Cutoff	Number	Buprenorphine	Lay person results	The
-------------	--------	---------------	--------------------	-----

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

	ofsamples	Concentration by GC/MS(ng/mL)	No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	2.5	0	20	100%
-50% Cutoff	20	5	0	20	100%
-25% Cutoff	20	7.5	2	18	90%
+25% Cutoff	20	12.5	18	2	90%
+50% Cutoff	20	15	20	0	100%
+75% Cutoff	20	17.5	20	0	100%

Comparison between GC/MS and Lay Person Results (Methadone Strip)

% of Cutoff	Numberofsamples	MethadoneConcentration by GC/MS(ng/mL)	Lay person resultsNo. ofPositive	Lay person resultsNo. ofNegative	Thepercentage ofcorrect results(%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

Comparison between GC/MS and Lay Person Results (Methadone Cassette)

% of Cutoff	Numberofsamples	MethadoneConcentration 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	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

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

% of Cutoff	Number	Methadone	Lay person results	The
-------------	--------	-----------	--------------------	-----

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

	ofsamples	Concentration by GC/MS(ng/mL)	No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100%Cutoff	20	0	0	20	100%
-75%Cutoff	20	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	1	19	95%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	100%

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

% of Cutoff	Numberofsamples	MethadoneConcentration by GC/MS(ng/mL)	Lay person results		The
			No. ofPositive	No. ofNegative	percentage ofcorrect results(%)
-100% Cutoff	20	0	0	20	100%
-75% Cutoff	20	75	0	20	100%
-50% Cutoff	20	150	0	20	100%
-25% Cutoff	20	225	2	18	90%
+25% Cutoff	20	375	19	1	95%
+50% Cutoff	20	450	20	0	100%
+75% Cutoff	20	525	20	0	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 each package insert and the scores revealed a reading Grade Level of 7.

1. 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 Healgen Secobarbital Test, Healgen Buprenorphine Test and Healgen Methadone Test are substantially equivalent to the predicate.

Regulation Number and Section

§ 862.3150 Barbiturate test system.

(a)
Identification. A barbiturate test system is a device intended to measure barbiturates, a class of hypnotic and sedative drugs, in serum, urine, and gastric contents. Measurements obtained by this device are used in the diagnosis and treatment of barbiturate use or overdose and in monitoring levels of barbiturate to ensure appropriate therapy.(b)
Classification. Class II (special controls). A barbiturate 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).