ARK EDDP Assay

K182779 · Ark Diagnostics, Inc. · DJR · Nov 21, 2018 · Clinical Toxicology

Device Facts

• Record ID: K182779
• Device Name: ARK EDDP Assay
• Applicant: Ark Diagnostics, Inc.
• Product Code: DJR · Clinical Toxicology
• Decision Date: Nov 21, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3620
• Device Class: Class 2

Intended Use

The ARK EDDP Assay is an immunoassay intended for the qualitative and/or semiquantitative determination of EDDP in human urine at cutoff concentrations of 100 ng/mL and 300 ng/mL. The assay is intended for use in laboratories with automated clinical chemistry analyzers. This in vitro diagnostic device is for prescription use only. The semiquantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures. The ARK EDDP Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.

Device Story

ARK EDDP Assay is a homogeneous enzyme immunoassay for detecting EDDP in human urine. Input: urine specimen. Principle: competitive binding between EDDP in specimen and EDDP-labeled recombinant G6PDH enzyme for antibody binding sites. Output: absorbance change measured spectrophotometrically at 340 nm; enzyme activity is directly proportional to EDDP concentration. Used in clinical laboratories on automated chemistry analyzers by laboratory personnel. Provides preliminary results; requires confirmation by GC/MS or LC-MS/MS. Clinical decision-making relies on confirmed results; aids in identifying EDDP presence for clinical or quality control purposes.

Clinical Evidence

Bench testing only. Precision evaluated per CLSI EP05-A3 (N=160 per level). Analytical recovery (95.1-107.9%) and specificity tested against structurally related and unrelated compounds. Interference studies (endogenous substances, pH, specific gravity) showed no interference. Method comparison performed on 109 clinical urine specimens against GC/MS reference method, showing high concordance at 100 ng/mL and 300 ng/mL cutoffs.

Technological Characteristics

Homogeneous enzyme immunoassay. Reagents: R1 (anti-EDDP rabbit antibody with substrate) and R2 (EDDP derivative labeled with bacterial recombinant G6PDH). Detection: spectrophotometric absorbance at 340 nm. Platform: automated clinical chemistry analyzers. Storage: 2-8°C. Preservative: sodium azide.

Indications for Use

Indicated for qualitative and/or semiquantitative determination of EDDP in human urine at 100 ng/mL and 300 ng/mL cutoffs. Intended for prescription use in clinical laboratories using automated chemistry analyzers.

Regulatory Classification

Identification

A methadone test system is a device intended to measure methadone, an addictive narcotic pain-relieving drug, in serum and urine. Measurements obtained by this device are used in the diagnosis and treatment of methadone use or overdose and to determine compliance with regulations in methadone maintenance treatment.

Special Controls

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

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logos of the Department of Health & Human Services and the Food and Drug Administration (FDA). The Department of Health & Human Services logo is on the left, and the FDA logo is on the right. The FDA logo is a blue square with the letters "FDA" in white, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.

November 21, 2018

ARK Diagnostics, Inc. Cherry Mun Manager, Quality and Regulatory Affairs 48089 Fremont Boulevard Fremont, California 94538

Re: k182779

Trade/Device Name: ARK EDDP Assay Regulation Number: 21 CFR 862.3620 Regulation Name: Methadone test system Regulatory Class: Class II Product Code: DJR Dated: September 28, 2018 Received: October 1, 2018

#### Dear Cherry Mun:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part

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801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/CombinationProducts/GuidanceRegulatoryInformation/ucm597488.htm); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and

Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

# Paula Caposino -S

for Courtnev 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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| Form Approved: OMB No. 0910-0120 | |
|----------------------------------|--|
| Expiration Date: 06/30/2020 | |
| See PRA Statement below. | |

DEPARTMENT OF HEALTH AND HUMAN SERVICES

Food and Drug Administration

**Indications for Use**

| 510(k) Number (if known) | k182779 |
|--------------------------|----------------|
| Device Name | ARK EDDP Assay |

**Indications for Use (Describe)**

The ARK EDDP Assay is an immunoassay intended for the qualitative and/or semiquantitative determination of EDDP in
human urine at cutoff concentrations of 100 ng/mL and 300 ng/mL. The assay is intended for use in laboratories with
automated clinical chemistry analyzers. This in vitro diagnostic device is for prescription use only.

The semiquantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the
specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or
Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality
control procedures.

The ARK EDDP Assay provides only a preliminary analytical test result. A more specific alternative chemical method
must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS)
or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical
consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary
test result is positive.

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

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# 510(k) SUMMARY

This 510(k) Summary of Safety and Effectiveness information is being submitted in accordance with the requirements of Safe Medical Device Act of 1990 and 21 CFR 807.92.

The assigned 510(k) number is K182779.

| 807.92 (a)(1): Name: | ARK Diagnostics, Inc. |
|-----------------------------|--------------------------------------------------------------------------|
| Address: | 48089 Fremont Blvd<br>Fremont, CA 94538 USA |
| Owner Operator Number: | 10027663 |
| Establishment Registration: | 3005755244 |
| Phone: | [phone] |
| FAX: | [phone] |
| Contact: | Cherry Mun – [phone]<br>Manager of Quality and Regulatory Affairs |

Date Prepared: November 19th, 2018

#### 807.92 (a)(2): Device Name – Trade Name, Common Name, and Classification

| Trade Name: | ARK™ EDDP Assay |
|--------------|-------------------------------------------------------|
| Common Name: | Homogeneous Enzyme Immunoassay, Methadone Test System |

Classification:

| Product Code | Classification | Regulation Section | Panel |
|--------------|----------------|------------------------------------------|--------------------|
| DJR | Class II | 21 CFR 862.3620<br>Methadone Test System | Toxicology<br>(91) |

## 807.92 (a)(3): Identification of the Legally Marketed Predicate Device

Immunalysis EDDP Specific Urine Enzyme Immunoassay (K151395)

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## 807.92 (a)(4): Device Description

The ARK EDDP Assay is a homogeneous enzyme immunoassay technique used for the analysis of EDDP in human urine. The assay is based on competition between EDDP in the specimen and EDDP labeled with recombinant glucose-6-phosphate dehydrogenase (rG6PDH) for antibody binding sites. As the latter binds antibody, enzyme activity decreases. In the presence of EDDP from the specimen, enzyme activity increases and is directly related to the EDDP concentration. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH in the presence of glucose-6-phosphate (G6P), resulting in an absorbance change that is measured spectrophotometrically. Endogenous G6PDH does not interfere because the coenzyme NAD functions only with the bacterial enzyme used in the assay.

The ARK EDDP Assay consists of reagents R1 anti-EDDP rabbit antibody with substrate and R2 EDDP derivative labeled with bacterial recombinant G6PDH enzyme.

## 807.92 (a)(5): Intended Use / Indications for Use

## ARK EDDP Assay

The ARK EDDP Assay is an immunoassay intended for the qualitative and/or semiquantitative determination of EDDP in human urine at cutoff concentrations of 100 ng/mL and 300 ng/mL. The assay is intended for use in laboratories with automated clinical chemistry analyzers. This in vitro diagnostic device is for prescription use only.

The semiquantitative mode is for the purpose of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method, such as Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS), or (2) permitting laboratories to establish quality control procedures.

The ARK EDDP Assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed positive analytical result. Gas Chromatography/Mass Spectrometry (GC/MS) or Liquid Chromatography/tandem Mass Spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug test result, particularly when the preliminary test result is positive.

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## 807.92 (a)(6): Technological Similarities and Differences to the Predicate

## SUBSTANTIAL EQUIVALENCE COMPARATIVE TABLE

## Comparison between the Immunalysis EDDP Specific Urine Enzyme Immunoassay and the ARK™ EDDP Assay

| Characteristic | Predicate Device<br>Immunalysis EDDP Specific Urine<br>Enzyme Immunoassay (K151395) | Candidate Device<br>ARKTM EDDP Assay |
|-----------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------|
| Similarities | | |
| Test System | Homogenous enzyme immunoassay<br>(EIA) | Same |
| Intended Use | For the qualitative and semiquantitative<br>determination of EDDP in human urine;<br>For in vitro diagnostic use | Same |
| Sample Matrix | Human urine | Same |
| User Environment | Clinical laboratories; Prescription use<br>only | Same |
| Mass Spectrometry<br>Confirmation | Required to confirm preliminary positive<br>analytical results | Same |
| Platform Required | Automated clinical chemistry analyzer | Same |
| Reagents Form | Liquid - Ready to use | Same |
| Reagent Materials | Two (2) reagent system:<br>Antibody/substrate reagent (antibodies to<br>EDDP) and enzyme labeled conjugate<br>(EDDP derivative labeled with enzyme)<br>Sodium azide preservative | Same |
| Storage | 2-8°C until expiration date | Same |
| Measured Analyte | EDDP | Same |
| Detection | Absorbance change measured<br>spectrophotometrically at 340 nm | Same |
| Characteristic | Predicate Device<br>Immunalysis EDDP Specific Urine<br>Enzyme Immunoassay (K151395) | Candidate Device<br>ARKTM EDDP Assay |
| Differences | | |
| Cutoff Levels | 100 ng/mL, 300 ng/mL and 1000 ng/mL | 100 ng/mL and 300 ng/mL |

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## 807.92 (b)(1) and 807.92 (b)(2): Brief Description of Nonclinical and Clinical Data

The following performance characteristics were obtained on the Beckman Coulter AU680® automated clinical chemistry analyzer.

#### Precision

Precision studies were performed using CLSI EP05-A3 as a guideline. Drug-free, negative human urine was supplemented with EDDP (0.0 to 200.0 ng/mL for the 100 ng/mL cutoff and 0.0 to 600.0 for the 300 ng/mL cutoff). Each level was assayed in quadruplicate twice a day for 20 days (N=160) and evaluated qualitatively and semiquantitatively. Results are summarized in the tables below.

| Human Urine<br>(ng/mL) | Relative %<br>Cutoff | # of Results | Qualitative<br>Precision Results |
|------------------------|----------------------|--------------|----------------------------------|
| 0.0 | -100 | 160 | 160 Negative |
| 25.0 | -75 | 160 | 160 Negative |
| 50.0 | -50 | 160 | 160 Negative |
| 75.0 | -25 | 160 | 160 Negative |
| 100.0 | Cutoff | 160 | 123 Negative;<br>37 Positive |
| 125.0 | +25 | 160 | 160 Positive |
| 150.0 | +50 | 160 | 160 Positive |
| 175.0 | +75 | 160 | 160 Positive |
| 200.0 | +100 | 160 | 160 Positive |

## Qualitative Precision - 100 ng/mL Cutoff

## Semiquantitative Precision – 100 ng/mL Cutoff

| Human Urine<br>(ng/mL) | Relative %<br>Cutoff | # of Results | Mean<br>(ng/mL) | Semiquantitative<br>Precision Results |
|------------------------|----------------------|--------------|-----------------|---------------------------------------|
| 0.0 | -100 | 160 | 0.3 | 160 Negative |
| 25.0 | -75 | 160 | 22.6 | 160 Negative |
| 50.0 | -50 | 160 | 47.7 | 160 Negative |
| 75.0 | -25 | 160 | 72.2 | 160 Negative |
| 100.0 | Cutoff | 160 | 98.1 | 114 Negative;<br>46 Positive |
| 125.0 | +25 | 160 | 125.3 | 160 Positive |
| 150.0 | +50 | 160 | 145.1 | 160 Positive |
| 175.0 | +75 | 160 | 169.4 | 160 Positive |
| 200.0 | +100 | 160 | 190.7 | 160 Positive |

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## Qualitative Precision – 300 ng/mL Cutoff

| Human Urine<br>(ng/mL) | Relative %<br>Cutoff | # of Results | Qualitative<br>Precision Results |
|------------------------|----------------------|--------------|----------------------------------|
| 0.0 | -100 | 160 | 160 Negative |
| 75.0 | -75 | 160 | 160 Negative |
| 150.0 | -50 | 160 | 160 Negative |
| 225.0 | -25 | 160 | 160 Negative |
| 300.0 | Cutoff | 160 | 57 Negative;<br>103 Positive |
| 375.0 | +25 | 160 | 160 Positive |
| 450.0 | +50 | 160 | 160 Positive |
| 525.0 | +75 | 160 | 160 Positive |
| 600.0 | +100 | 160 | 160 Positive |

## Semiquantitative Precision – 300 ng/mL Cutoff

| Human Urine<br>(ng/mL) | Relative %<br>Cutoff | # of Results | Mean<br>(ng/mL) | Semiquantitative<br>Precision Results |
|------------------------|----------------------|--------------|-----------------|---------------------------------------|
| 0.0 | -100 | 160 | 0.3 | 160 Negative |
| 75.0 | -75 | 160 | 72.2 | 160 Negative |
| 150.0 | -50 | 160 | 145.1 | 160 Negative |
| 225.0 | -25 | 160 | 205.9 | 160 Negative |
| 300.0 | Cutoff | 160 | 298.8 | 85 Negative;<br>75 Positive |
| 375.0 | +25 | 160 | 381.4 | 160 Positive |
| 450.0 | +50 | 160 | 461.0 | 160 Positive |
| 525.0 | +75 | 160 | 539.8 | 160 Positive |
| 600.0 | +100 | 160 | 620.0 | 160 Positive |

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## Analytical Recovery

Recovery across the assay range was assessed using the semiquantitative mode. Drug-free, negative human urine was supplemented with EDDP (1100.0 ng/mL) and dilutions were made proportionally with drug-free human urine. EDDP concentrations ranged from 50.0 to 1000.0 ng/mL. At each level, percentage recovery was calculated based on the mean concentration (N=6) compared to the expected concentration. Results are summarized in the table below.

| Theoretical<br>Concentration<br>(ng/mL) | Mean<br>Concentration<br>(ng/mL) | Recovery<br>(%) |
|-----------------------------------------|----------------------------------|-----------------|
| 50.0 | 47.6 | 95.1 |
| 75.0 | 72.1 | 96.1 |
| 100.0 | 97.1 | 97.1 |
| 200.0 | 189.1 | 94.6 |
| 300.0 | 286.6 | 95.5 |
| 400.0 | 414.5 | 103.6 |
| 500.0 | 506.6 | 101.3 |
| 600.0 | 647.4 | 107.9 |
| 700.0 | 722.7 | 103.2 |
| 800.0 | 800.6 | 100.1 |
| 900.0 | 880.8 | 97.9 |
| 1000.0 | 955.8 | 95.6 |

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#### Analytical Specificity

All compounds tested were added to drug-free, negative human urine and tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

The cross-reactivity of the following structurally related compounds was evaluated by spiking these compounds into drug-free, negative human urine to determine the minimum concentration that would give a positive result approximately equivalent to the 100 ng/mL and 300 ng/mL EDDP cutoffs. These concentrations were used to determine the percent cross-reactivity according to the formula:

% Cross-reactivity = (Cutoff concentration / Lowest concentration of cross-reactant causing a positive result) X 100

For compounds that did not produce a positive result, the highest concentration tested was used to calculate percent cross-reactivity.

| Compound | Concentration<br>Tested<br>(ng/mL) | Semiquantitative<br>Mode Result<br>(Positive/Negative) | Qualitative Mode<br>Result<br>(Positive/Negative) | Cross-reactivity<br>(%) |
|-----------------|------------------------------------|--------------------------------------------------------|---------------------------------------------------|-------------------------|
| EDDP | 100 | Positive | Positive | 100 |
| Methadone | 2,000,000 | Negative | Negative | <0.005 |
| EMDP | 400,000 | Negative | Negative | <0.025 |
| Chlorpromazine | 100,000 | Negative | Negative | <0.1 |
| Diphenhydramine | 100,000 | Negative | Negative | <0.1 |
| Methylphenidate | 100,000 | Negative | Negative | <0.1 |
| Doxylamine | 100,000 | Negative | Negative | <0.1 |

#### Structurally Related Compounds - 100 ng/mL Cutoff

#### Structurally Related Compounds - 300 ng/mL Cutoff

| Compound | Concentration<br>Tested<br>(ng/mL) | Semiquantitative<br>Mode Result<br>(Positive/Negative) | Qualitative Mode<br>Result<br>(Positive/Negative) | Cross-reactivity<br>(%) |
|-----------------|------------------------------------|--------------------------------------------------------|---------------------------------------------------|-------------------------|
| EDDP | 300 | Positive | Positive | 100 |
| Methadone | 4,500,000 | Negative | Negative | <0.007 |
| EMDP | 1,000,000 | Negative | Negative | <0.03 |
| Chlorpromazine | 100,000 | Negative | Negative | <0.3 |
| Diphenhydramine | 100,000 | Negative | Negative | <0.3 |
| Methylphenidate | 100,000 | Negative | Negative | <0.3 |
| Doxylamine | 100,000 | Negative | Negative | <0.3 |

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#### Interference

#### Structurally Unrelated Compounds – 100 ng/mL Cutoff

High concentrations of the following structurally unrelated compounds were added into urine spiked with EDDP (± 25% of the 100 ng/mL cutoff concentration) and tested with the ARK EDDP Assay in both qualitative and semiquantitative modes. The substances listed at the concentrations below did not yield a false result relative to the 100 ng/mL cutoff.

| Compound | Concentration<br>Tested<br>(ng/mL) | Spiked EDDP Level | |
|--------------------------------------|------------------------------------|---------------------------|----------------------------|
| | | 75 ng/mL<br>(-25% Cutoff) | 125 ng/mL<br>(+25% Cutoff) |
| 4-bromo 2-5, dimethoxyphenethylamine | 100,000 | Negative | Positive |
| Acetaminophen | 500,000 | Negative | Positive |
| Acetylsalicylic Acid | 500,000 | Negative | Positive |
| 6-Acetylcodeine | 100,000 | Negative | Positive |
| 6-Acetylmorphine | 100,000 | Negative | Positive |
| Alprazolam | 100,000 | Negative | Positive |
| 7-Aminoclonazepam | 100,000 | Negative | Positive |
| 7-Aminoflunitrazepam | 100,000 | Negative | Positive |
| 7-Aminonitrazepam | 100,000 | Negative | Positive |
| Amitriptyline | 100,000 | Negative | Positive |
| Amobarbital | 100,000 | Negative | Positive |
| S-(+)-Amphetamine | 100,000 | Negative | Positive |
| Benzylpiperazine | 100,000 | Negative | Positive |
| Bromazepam | 100,000 | Negative | Positive |
| Buprenorphine | 100,000 | Negative | Positive |
| Bupropion | 100,000 | Negative | Positive |
| Butabarbital | 100,000 | Negative | Positive |
| Butalbital | 100,000 | Negative | Positive |
| Caffeine | 500,000 | Negative | Positive |
| Cannabidiol | 100,000 | Negative | Positive |
| Cannabinol | 100,000 | Negative | Positive |
| Carbamazepine | 100,000 | Negative | Positive |
| Carisoprodol | 100,000 | Negative | Positive |
| Chlordiazepoxide | 100,000 | Negative | Positive |
| Cis-Tramadol | 100,000 | Negative | Positive |
| Clobazam | 100,000 | Negative | Positive |
| Clomipramine | 100,000 | Negative | Positive |
| Clonazepam | 100,000 | Negative | Positive |
| Clozapine | 100,000 | Negative | Positive |
| Codeine | 100,000 | Negative | Positive |
| Cotinine | 100,000 | Negative | Positive |
| Cyclobenzaprine | 100,000 | Negative | Positive |
| Dehydronorketamine | 100,000 | Negative | Positive |
| Demoxepam | 100,000 | Negative | Positive |
| Desipramine | 100,000 | Negative | Positive |
| Desalkylflurazepam | 100,000 | Negative | Positive |
| Dextromethorphan | 100,000 | Negative | Positive |

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| | Concentration | Spiked EDDP Level | |
|---------------------------|---------------|-------------------|---------------|
| Compound | Tested | 75 ng/mL | 125 ng/mL |
| | (ng/mL) | (-25% Cutoff) | (+25% Cutoff) |
| Diazepam | 100,000 | Negative | Positive |
| Digoxin | 100,000 | Negative | Positive |
| Dihydrocodeine | 100,000 | Negative | Positive |
| Δ9 THC | 100,000 | Negative | Positive |
| Doxepin | 100,000 | Negative | Positive |
| 1R,2S (-) Ephedrine | 100,000 | Negative | Positive |
| 1S,2R (+) Ephedrine | 100,000 | Negative | Positive |
| Ethyl-ß-D-Glucuronide | 100,000 | Negative | Positive |
| Ethylmorphine | 100,000 | Negative | Positive |
| (S-)-Fenfluramine | 100,000 | Negative | Positive |
| (R+)-Fenfluramine | 100,000 | Negative | Positive |
| Fentanyl | 100,000 | Negative | Positive |
| Flunitrazepam | 100,000 | Negative | Positive |
| Fluoxetine | 100,000 | Negative | Positive |
| Flurazepam | 100,000 | Negative | Positive |
| Haloperidol | 100,000 | Negative | Positive |
| Heroin | 100,000 | Negative | Positive |
| Hexobarbital | 100,000 | Negative | Positive |
| Hydrocodone | 100,000 | Negative | Positive |
| Hydromorphone | 100,000 | Negative | Positive |
| 11-hydroxy-Δ9 THC | 100,000 | Negative | Positive |
| Ibuprofen | 500,000 | Negative | Positive |
| Imipramine | 100,000 | Negative | Positive |
| Ketamine | 100,000 | Negative | Positive |
| Lamotrigine | 100,000 | Negative | Positive |
| Levorphanol Tartrate | 100,000 | Negative | Positive |
| Lidocaine | 100,000 | Negative | Positive |
| Lorazepam | 100,000 | Negative | Positive |
| Lorazepam Glucuronide | 50,000 | Negative | Positive |
| Lormetrazepam | 100,000 | Negative | Positive |
| LSD | 100,000 | Negative | Positive |
| Maprotiline | 100,000 | Negative | Positive |
| (+)-MDA | 100,000 | Negative | Positive |
| MDEA | 100,000 | Negative | Positive |
| MDMA | 100,000 | Negative | Positive |
| Meperidine | 100,000 | Negative | Positive |
| Meprobamate | 100,000 | Negative | Positive |
| S(+)-Methamphetamine | 100,000 | Negative | Positive |
| Methaquolone | 100,000 | Negative | Positive |
| Methoxetamine | 100,000 | Negative | Positive |
| Methylone | 100,000 | Negative | Positive |
| Midazolam | 100,000 | Negative | Positive |
| Morphine | 100,000 | Negative | Positive |
| Morphine-3B-D-Glucuronide | 100,000 | Negative | Positive |
| Morphine-6ß-D-Glucuronide | 50,000 | Negative | Positive |
| N-Desmethyltapentadol | 100,000 | Negative | Positive |
| Nalorphine | 100,000 | Negative | Positive |

ARK Diagnostics, Inc. – 510(k) Summary ARK EDDP Assay

Page 5-9 of 5-19

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| Compound | Concentration<br>Tested<br>(ng/mL) | Spiked EDDP Level | |
|---------------------------------|------------------------------------|---------------------------|----------------------------|
| | | 75 ng/mL<br>(-25% Cutoff) | 125 ng/mL<br>(+25% Cutoff) |
| Naloxone | 100,000 | Negative | Positive |
| Naltrexone | 100,000 | Negative | Positive |
| Naproxen | 100,000 | Negative | Positive |
| Nitrazepam | 100,000 | Negative | Positive |
| 11-nor-9-carboxy-Δ9-THC | 100,000 | Negative | Positive |
| Norbuprenorphine | 50,000 | Negative | Positive |
| Norcodeine | 100,000 | Negative | Positive |
| Nordiazepam | 100,000 | Negative | Positive |
| Norketamine | 100,000 | Negative | Positive |
| Normorphine | 100,000 | Negative | Positive |
| Norpropoxyphene | 100,000 | Negative | Positive |
| Norpseudoephedrine | 100,000 | Negative | Positive |
| Nortriptyline | 100,000 | Negative | Positive |
| Olanzapine | 100,000 | Negative | Positive |
| Oxazepam | 100,000 | Negative | Positive |
| Oxycodone | 100,000 | Negative | Positive |
| Oxymorphone | 100,000 | Negative | Positive |
| PCP | 100,000 | Negative | Positive |
| Pentazocine | 100,000 | Negative | Positive |
| Pentobarbital | 100,000 | Negative | Positive |
| Phenobarbital | 100,000 | Negative | Positive |
| Phentermine | 100,000 | Negative | Positive |
| Phenylephedrine | 100,000 | Negative | Positive |
| Phenylpropanolamine | 100,000 | Negative | Positive |
| Phenytoin | 100,000 | Negative | Positive |
| PMA | 100,000 | Negative | Positive |
| Prazepam | 100,000 | Negative | Positive |
| Propoxyphene | 100,000 | Negative | Positive |
| Propranolol | 100,000 | Negative | Positive |
| Protriptyline | 100,000 | Negative | Positive |
| R,R (+)- Pseudoephedrine | 100,000 | Negative | Positive |
| S,S (-)- Pseudoephedrine | 100,000 | Negative | Positive |
| Ranitidine | 100,000 | Negative | Positive |
| Ritalinic Acid | 100,000 | Negative | Positive |
| Salicylic Acid | 100,000 | Negative | Positive |
| Secobarbital | 100,000 | Negative | Positive |
| Sertraline | 100,000 | Negative | Positive |
| Sufentanil Citrate | 50,000 | Negative | Positive |
| Tapentadol | 100,000 | Negative | Positive |
| Temazepam | 100,000 | Negative | Positive |
| Theophylline | 100,000 | Negative | Positive |
| Thioridazine | 100,000 | Negative | Positive |
| Trazodone | 100,000 | Negative | Positive |
| Triazolam | 100,000 | Negative | Positive |
| Trifluoromethylphenylpiperazine | 100,000 | Negative | Positive |
| Trimipramine | 100,000 | Negative | Positive |
| Venlafaxine | 100,000 | Negative | Positive |

ARK Diagnostics, Inc. – 510(k) Summary ARK EDDP Assay

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

| | Concentration | Spiked EDDP Level | |
|-------------------|-------------------|---------------------------|----------------------------|
| Compound | Tested<br>(ng/mL) | 75 ng/mL<br>(-25% Cutoff) | 125 ng/mL<br>(+25% Cutoff) |
| Verapamil | 100,000 | Negative | Positive |
| Zolpidem Tartrate | 100,000 | Negative | Positive |

## Structurally Unrelated Compounds – 300 ng/mL Cutoff

High concentrations of the following structurally unrelated compounds were added into urine spiked with EDDP (± 25% of the 300 ng/mL cutoff concentration) and tested with the ARK EDDP Assay in both qualitative and semiquantitative modes. The substances listed at the concentrations below did not yield a false result relative to the 300 ng/mL cutoff.

| Compound | Concentration<br>Tested<br>(ng/mL) | Spiked EDDP Level | |
|--------------------------------------|------------------------------------|----------------------------|----------------------------|
| | | 225 ng/mL<br>(-25% Cutoff) | 375 ng/mL<br>(+25% Cutoff) |
| 4-bromo 2-5, dimethoxyphenethylamine | 100,000 | Negative | Positive |
| Acetaminophen | 500,000 | Negative | Positive |
| Acetylsalicylic Acid | 500,000 | Negative | Positive |
| 6-Acetylcodeine | 100,000 | Negative | Positive |
| 6-Acetylmorphine | 100,000 | Negative | Positive |
| Alprazolam | 100,000 | Negative | Positive |
| 7-Aminoclonazepam | 100,000 | Negative | Positive |
| 7-Aminoflunitrazepam | 100,000 | Negative | Positive |
| 7-Aminonitrazepam | 100,000 | Negative | Positive |
| Amitriptyline | 100,000 | Negative | Positive |
| Amobarbital | 100,000 | Negative | Positive |
| S-(+)-Amphetamine | 100,000 | Negative | Positive |
| Benzylpiperazine | 100,000 | Negative | Positive |
| Bromazepam | 100,000 | Negative | Positive |
| Buprenorphine | 100,000 | Negative | Positive |
| Bupropion | 100,000 | Negative | Positive |
| Butabarbital | 100,000 | Negative | Positive |
| Butalbital | 100,000 | Negative | Positive |
| Caffeine | 500,000 | Negative | Positive |
| Cannabidiol | 100,000 | Negative | Positive |
| Cannabinol | 100,000 | Negative | Positive |
| Carbamazepine | 100,000 | Negative | Positive |
| Carisoprodol | 100,000 | Negative | Positive |
| Chlordiazepoxide | 100,000 | Negative | Positive |
| Cis-Tramadol | 100,000 | Negative | Positive |
| Clobazam | 100,000 | Negative | Positive |
| Clomipramine | 100,000 | Negative | Positive |
| Clonazepam | 100,000 | Negative | Positive |
| Clozapine | 100,000 | Negative | Positive |
| Codeine | 100,000 | Negative | Positive |
| Cotinine | 100,000 | Negative | Positive |
| Cyclobenzaprine | 100,000 | Negative | Positive |
| Dehydronorketamine | 100,000 | Negative | Positive |
| | Concentration | Spiked EDDP Level | |
| Compound | Tested (ng/mL) | 225 ng/mL (-25% Cutoff) | 375 ng/mL (+25% Cutoff) |
| Demoxepam | 100,000 | Negative | Positive |
| Desipramine | 100,000 | Negative | Positive |
| Desalkylflurazepam | 100,000 | Negative | Positive |
| Dextromethorphan | 100,000 | Negative | Positive |
| Diazepam | 100,000 | Negative | Positive |
| Digoxin | 100,000 | Negative | Positive |
| Dihydrocodeine | 100,000 | Negative | Positive |
| Δ9 THC | 100,000 | Negative | Positive |
| Doxepin | 100,000 | Negative | Positive |
| 1R,2S (-) Ephedrine | 100,000 | Negative | Positive |
| 1S,2R (+) Ephedrine | 100,000 | Negative | Positive |
| Ethyl-β-D-Glucuronide | 100,000 | Negative | Positive |
| Ethylmorphine | 100,000 | Negative | Positive |
| (S-)-Fenfluramine | 100.000 | Negative | Positive |
| (R+)-Fenfluramine | 100,000 | Negative | Positive |
| Fentanyl | 100,000 | Negative | Positive |
| Flunitrazepam | 100,000 | Negative | Positive |
| Fluoxetine | 100,000 | Negative | Positive |
| Flurazepam | 100,000 | Negative | Positive |
| Haloperidol | 100,000 | Negative | Positive |
| Heroin | 100,000 | Negative | Positive |
| Hexobarbital | 100,000 | Negative | Positive |
| Hydrocodone | 100,000 | Negative | Positive |
| Hydromorphone | 100,000 | Negative | Positive |
| 11-hydroxy-Δ9 THC | 100,000 | Negative | Positive |
| Ibuprofen | 500,000 | Negative | Positive |
| Imipramine | 100,000 | Negative | Positive |
| Ketamine | 100,000 | Negative | Positive |
| Lamotrigine | 100,000 | Negative | Positive |
| Levorphanol Tartrate | 100,000 | Negative | Positive |
| Lidocaine | 100,000 | Negative | Positive |
| Lorazepam | 100.000 | Negative | Positive |
| Lorazepam Glucuronide | 50,000 | Negative | Positive |
| Lormetrazepam | 100,000 | Negative | Positive |
| LSD | 100,000 | Negative | Positive |
| Maprotiline | 100,000 | Negative | Positive |
| (+)-MDA | 100,000 | Negative | Positive |
| MDEA | 100,000 | Negative | Positive |
| MDMA | 100,000 | Negative | Positive |
| Meperidine | 100,000 | Negative | Positive |
| Meprobamate | 100,000 | Negative | Positive |
| S(+)-Methamphetamine | 100,000 | Negative | Positive |
| Methaquolone | 100,000 | Negative | Positive |
| Methoxetamine | 100,000 | Negative | Positive |
| Methylone | 100,000 | Negative | Positive |
| Midazolam | 100,000 | Negative | Positive |
| Morphine | 100,000 | Negative | Positive |
| Compound | Concentration<br>Tested<br>(ng/mL) | Spiked EDDP Level | |
| | | 225 ng/mL<br>(-25% Cutoff) | 375 ng/mL<br>(+25% Cutoff) |
| Morphine-3β-D-Glucuronide | 100,000 | Negative | Positive |
| Morphine-6β-D-Glucuronide | 50,000 | Negative | Positive |
| N-Desmethyltapentadol | 100,000 | Negative | Positive |
| Nalorphine | 100,000 | Negative | Positive |
| Naloxone | 100,000 | Negative | Positive |
| Naltrexone | 100,000 | Negative | Positive |
| Naproxen | 100,000 | Negative | Positive |
| Nitrazepam | 100,000 | Negative | Positive |
| 11-nor-9-carboxy-Δ9-THC | 100,000 | Negative | Positive |
| Norbuprenorphine | 50,000 | Negative | Positive |
| Norcodeine | 100,000 | Negative | Positive |
| Nordiazepam | 100,000 | Negative | Positive |
| Norketamine | 100,000 | Negative | Positive |
| Normorphine | 100,000 | Negative | Positive |
| Norpropoxyphene | 75,000 | Negative | Positive |
| Norpseudoephedrine | 100,000 | Negative | Positive |
| Nortriptyline | 100,000 | Negative | Positive |
| Olanzapine | 100,000 | Negative | Positive |
| Oxazepam | 100,000 | Negative | Positive |
| Oxycodone | 100,000 | Negative | Positive |
| Oxymorphone | 100,000 | Negative | Positive |
| PCP | 50,000 | Negative | Positive |
| Pentazocine | 100,000 | Negative | Positive |
| Pentobarbital | 100,000 | Negative | Positive |
| Phenobarbital | 100,000 | Negative | Positive |
| Phentermine | 100,000 | Negative | Positive |
| Phenylephedrine | 100,000 | Negative | Positive |
| Phenylpropanolamine | 100,000 | Negative | Positive |
| Phenytoin | 100,000 | Negative | Positive |
| PMA | 100,000 | Negative | Positive |
| Prazepam | 100,000 | Negative | Positive |
| Propoxyphene | 100,000 | Negative | Positive |
| Propranolol | 100,000 | Negative | Positive |
| Protriptyline | 100,000 | Negative | Positive |
| R,R (+)- Pseudoephedrine | 100,000 | Negative | Positive |
| S,S (-)- Pseudoephedrine | 100,000 | Negative | Positive |
| Ranitidine | 100,000 | Negative | Positive |
| Ritalinic Acid | 100,000 | Negative | Positive |
| Salicylic Acid | 100,000 | Negative | Positive |
| Secobarbital | 100,000 | Negative | Positive |
| Sertraline | 100,000 | Negative | Positive |
| Sufentanil Citrate | 50,000 | Negative | Positive |
| Tapentadol | 100,000 | Negative | Positive |
| Temazepam | 100,000 | Negative | Positive |
| Theophylline | 100,000 | Negative | Positive |
| Thioridazine | 100,000 | Negative | Positive |
| Trazodone | 100,000 | Negative | Positive |
| Compound | Concentration<br>Tested<br>(ng/mL) | Spiked EDDP Level | |
| | | 225 ng/mL<br>(-25% Cutoff) | 375 ng/mL<br>(+25% Cutoff) |
| Triazolam | 100,000 | Negative | Positive |
| Trifluoromethylphenylpiperazine | 100,000 | Negative | Positive |
| Trimipramine | 100,000 | Negative | Positive |
| Venlafaxine | 100,000 | Negative | Positive |
| Verapamil | 100,000 | Negative | Positive |
| Zolpidem Tartrate | 100,000 | Negative | Positive |

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

ARK Diagnostics, Inc. – 510(k) Summary ARK EDDP Assay

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ARK Diagnostics, Inc. – 510(k) Summary ARK EDDP Assay

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#### Endogenous Substances – 100 ng/mL Cutoff

Interference studies were performed using CLSI EP07-A2 as a guideline. High concentrations of the following endogenous substances were added into urine spiked with EDDP (± 25% of the 100 ng/mL cutoff concentration). No interference was observed when tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

| | | Spiked EDDP Level | | |
|--------------------------|-------------------------|---------------------------|----------------------------|--|
| Compound | Concentration<br>Tested | 75 ng/mL<br>(-25% Cutoff) | 125 ng/mL<br>(+25% Cutoff) | |
| Acetone | 1000 mg/dL | Negative | Positive | |
| Ascorbic Acid | 1500 mg/dL | Negative | Positive | |
| Bilirubin – Conjugated | 2 mg/dL | Negative | Positive | |
| Bilirubin – Unconjugated | 2 mg/dL | Negative | Positive | |
| Boric Acid | 1% w/v | Negative | Positive | |
| Creatinine | 500 mg/dL | Negative | Positive | |
| Ethanol | 1000 mg/dL | Negative | Positive | |
| Galactose | 10 mg/dL | Negative | Positive | |
| Gamma Globulin | 500 mg/dL | Negative | Positive | |
| Glucose | 2000 mg/dL | Negative | Positive | |
| Hemoglobin | 300 mg/dL | Negative | Positive | |
| Human Albumin | 500 mg/dL | Negative | Positive | |
| Oxalic Acid | 100 mg/dL | Negative | Positive | |
| Riboflavin | 7.5 mg/dL | Negative | Positive | |
| Sodium Azide | 1% w/v | Negative | Positive | |
| Sodium Chloride | 6000 mg/dL | Negative | Positive | |
| Sodium Fluoride | 1% w/v | Negative | Positive | |
| Urea | 6000 mg/dL | Negative | Positive | |

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

#### Endogenous Substances – 300 ng/mL Cutoff

Interference studies were performed using CLSI EP07-A2 as a guideline. High concentrations of the following endogenous substances were added into urine spiked with EDDP (± 25% of the 300 ng/mL cutoff concentration). No interference was observed when tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

| Compound | Concentration<br>Tested | Spiked EDDP Level | |
|--------------------------|-------------------------|----------------------------|----------------------------|
| | | 225 ng/mL<br>(-25% Cutoff) | 375 ng/mL<br>(+25% Cutoff) |
| Acetone | 1000 mg/dL | Negative | Positive |
| Ascorbic Acid | 1500 mg/dL | Negative | Positive |
| Bilirubin – Conjugated | 2 mg/dL | Negative | Positive |
| Bilirubin – Unconjugated | 2 mg/dL | Negative | Positive |
| Boric Acid | 1% w/v | Negative | Positive |
| Creatinine | 500 mg/dL | Negative | Positive |
| Ethanol | 1000 mg/dL | Negative | Positive |
| Galactose | 10 mg/dL | Negative | Positive |
| Gamma Globulin | 500 mg/dL | Negative | Positive |
| Glucose | 2000 mg/dL | Negative | Positive |
| Hemoglobin | 300 mg/dL | Negative | Positive |
| Human Albumin | 500 mg/dL | Negative | Positive |
| Oxalic Acid | 100 mg/dL | Negative | Positive |
| Riboflavin | 7.5 mg/dL | Negative | Positive |
| Sodium Azide | 1% w/v | Negative | Positive |
| Sodium Chloride | 6000 mg/dL | Negative | Positive |
| Sodium Fluoride | 1% w/v | Negative | Positive |
| Urea | 6000 mg/dL | Negative | Positive |

#### Specific Gravity - 100 ng/mL Cutoff

Urine samples with specific gravity values ranging from 1.002 to 1.030 were tested in the presence of the two levels of EDDP at ± 25% of the 100 ng/mL cutoff concentration. No interference was observed when tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

| | Spiked EDDP Level | |
|------------------------|---------------------------|----------------------------|
| Compound | 75 ng/mL<br>(-25% Cutoff) | 125 ng/mL<br>(+25% Cutoff) |
| Specific Gravity 1.002 | Negative | Positive |
| Specific Gravity 1.004 | Negative | Positive |
| Specific Gravity 1.012 | Negative | Positive |
| Specific Gravity 1.018 | Negative | Positive |
| Specific Gravity 1.019 | Negative | Positive |
| Specific Gravity 1.026 | Negative | Positive |
| Specific Gravity 1.030 | Negative | Positive |

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

#### Specific Gravity - 300 ng/mL Cutoff

Urine samples with specific gravity values ranging from 1.002 to 1.030 were tested in the presence of the two levels of EDDP at ± 25% of the 300 ng/mL cutoff concentration. No interference was observed when tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

| | Spiked EDDP Level | |
|------------------------|----------------------------|----------------------------|
| Compound | 225 ng/mL<br>(-25% Cutoff) | 375 ng/mL<br>(+25% Cutoff) |
| Specific Gravity 1.002 | Negative | Positive |
| Specific Gravity 1.004 | Negative | Positive |
| Specific Gravity 1.012 | Negative | Positive |
| Specific Gravity 1.018 | Negative | Positive |
| Specific Gravity 1.019 | Negative | Positive |
| Specific Gravity 1.026 | Negative | Positive |
| Specific Gravity 1.030 | Negative | Positive |

#### pH – 100 ng/mL Cutoff

Urine samples with pH values from 3.0 to 11.0 were tested in the presence of the two levels of EDDP at ± 25% of the 100 ng/mL cutoff concentration. No interference was observed when tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

| | Spiked EDDP Level | |
|---------------|---------------------------|----------------------------|
| Compound | 75 ng/mL<br>(-25% Cutoff) | 125 ng/mL<br>(+25% Cutoff) |
| Urine pH 3.0 | Negative | Positive |
| Urine pH 4.0 | Negative | Positive |
| Urine pH 5.0 | Negative | Positive |
| Urine pH 6.0 | Negative | Positive |
| Urine pH 7.0 | Negative | Positive |
| Urine pH 8.0 | Negative | Positive |
| Urine pH 9.0 | Negative | Positive |
| Urine pH 10.0 | Negative | Positive |
| Urine pH 11.0 | Negative | Positive |

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

## pH – 300 ng/mL Cutoff

Urine samples with pH values from 3.0 to 11.0 were tested in the presence of the two levels of EDDP at ± 25% of the 300 ng/mL cutoff concentration. No interference was observed when tested with the ARK EDDP Assay in both qualitative and semiquantitative modes.

| | Spiked EDDP Level | |
|---------------|----------------------------|----------------------------|
| Compound | 225 ng/mL<br>(-25% Cutoff) | 375 ng/mL<br>(+25% Cutoff) |
| Urine pH 3.0 | Negative | Positive |
| Urine pH 4.0 | Negative | Positive |
| Urine pH 5.0 | Negative | Positive |
| Urine pH 6.0 | Negative | Positive |
| Urine pH 7.0 | Negative | Positive |
| Urine pH 8.0 | Negative | Positive |
| Urine pH 9.0 | Negative | Positive |
| Urine pH 10.0 | Negative | Positive |
| Urine pH 11.0 | Negative | Positive |

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

## Method Comparison

A total of one hundred nine (109) unaltered clinical human urine specimens that are not individually identifiable were analyzed for EDDP at the two cutoff levels with the ARK EDDP Assay in both qualitative and semiquantitative modes and the results were compared to GC/MS. The GC/MS confirmatory method was performed by a licensed reference laboratory. Results are summarized in the tables below.

#### Method Comparison - 100 ng/mL Cutoff

| ARK<br>Immunoassay<br>Result | Low Negative<br>Less than<br>50% below<br>the Cutoff<br>(< 50 ng/mL<br>by GC/MS) | Near Cutoff<br>Negative<br>Between 50%<br>below the<br>Cutoff and the<br>Cutoff<br>(50 — 99<br>ng/mL by<br>GC/MS) | Near Cutoff<br>Positive<br>Between the<br>Cutoff and<br>50% above<br>the Cutoff<br>(100 - 150<br>ng/mL by<br>GC/MS) | High Positive<br>Greater than<br>50% above<br>the Cutoff<br>(> 150 ng/mL<br>by GC/MS) |
|------------------------------|----------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Negative | 40 | 5 | 0 | 0 |
| Positive | 0 | 0 | 4 | 60 |

#### Method Comparison – 300 ng/mL Cutoff

| ARK<br>Immunoassay<br>Result | Low Negative<br>Less than<br>50% below<br>the Cutoff<br>(< 150 ng/mL<br>by GC/MS) | Near Cutoff<br>Negative<br>Between 50%<br>below the<br>Cutoff and the<br>Cutoff<br>(150 – 299<br>ng/mL by<br>GC/MS) | Near Cutoff<br>Positive<br>Between the<br>Cutoff and<br>50% above<br>the Cutoff<br>(300 - 450<br>ng/mL by<br>GC/MS) | High Positive<br>Greater than<br>50% above<br>the Cutoff<br>(> 450 ng/mL<br>by GC/MS) |
|------------------------------|-----------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------|---…