CEDIA Heroin Metabolite (6-AM) assay

K192943 · Microgenics Corporation · DJG · Dec 16, 2019 · Clinical Toxicology

Device Facts

• Record ID: K192943
• Device Name: CEDIA Heroin Metabolite (6-AM) assay
• Applicant: Microgenics Corporation
• Product Code: DJG · Clinical Toxicology
• Decision Date: Dec 16, 2019
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3650
• Device Class: Class 2

Intended Use

The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to be used in laboratories and provides a rapid analytical screening procedure to detect 6-Acetylmorphine in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This product is intended to be used by trained professionals only. The semi-quantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures. The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas chromatography/mass spectrometry (GC/MS) or Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method. Clinical and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.

Device Story

CEDIA Heroin Metabolite (6-AM) assay is a homogeneous enzyme immunoassay for urine drug screening. Input: human urine sample. Principle: competitive binding between sample 6-AM and 6-AM-conjugated Enzyme Donor (ED) for antibody binding sites; free ED-6-AM re-associates with Enzyme Acceptor (EA) to form active β-galactosidase; active enzyme cleaves substrate to produce color change measured spectrophotometrically. Output: qualitative or semi-quantitative 6-AM concentration. Used in clinical laboratories; operated by trained professionals on Horiba Pentra C400 analyzer. Output used by clinicians as preliminary screening result; requires confirmation by GC/MS or LC-MS/MS. Benefits: rapid identification of heroin metabolite presence to guide clinical/professional judgment.

Clinical Evidence

Bench testing only. Precision study (n=80) performed at cutoff, 25%, 50%, 75% above/below cutoff. Method comparison against LC-MS/MS using 103 patient samples showed 100% concordance. Specificity and cross-reactivity evaluated against structurally related/unrelated compounds and common interfering substances (pH, specific gravity, endogenous substances); no significant interference observed.

Technological Characteristics

Homogeneous enzyme immunoassay (CEDIA). Reagents: mouse monoclonal antibodies, Enzyme Acceptor, Enzyme Donor conjugated to 6-AM, chlorophenol red-β-D galactopyranoside. Spectrophotometric detection. Analyzed on Horiba Pentra C400. Storage 2–8°C. No specific software algorithm class stated; relies on standard clinical chemistry analyzer spectrophotometric measurement.

Indications for Use

Indicated for the qualitative and/or semi-quantitative detection of heroin metabolite (6-Acetylmorphine) in human urine at a 10 ng/mL cutoff. Intended for use by trained laboratory professionals as a preliminary screening procedure.

Regulatory Classification

Identification

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

Special Controls

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

Predicate Devices

• CEDIA Heroin Metabolite (6-AM) Assay (K173183)

Submission Summary (Full Text)

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December 16, 2019

Microgenics Corporation Sharron Penn Senior Regulatory Affairs Specialist 46500 Kato Road Fremont, CA 94538

Re: K192943

Trade/Device Name: CEDIA Heroin Metabolite (6-AM) Assay Regulation Number: 21 CFR 862.3650 Regulation Name: Opiate test system Regulatory Class: Class II Product Code: DJG Dated: October 17, 2019 Received: October 18, 2019

Dear Sharron Penn:

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.

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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 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/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) 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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). 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 (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Marianela Perez-Torres, M.T., Ph.D. Acting Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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# Indications for Use

510(k) Number (if known) K192943

Device Name CEDIA Heroin Metabolite (6-AM) Assay

#### Indications for Use (Describe)

The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to be used in laboratories and provides a rapid analytical screening procedure to detect 6-Acetylmorphine in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This product is intended to be used by trained professionals only.

The semi-quantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.

The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas chromatography/ mass spectrometry (GC/MS) or Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.

Clinical and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only

| Type of Use (Select one or both, as applicable) | |
|-------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------|
| | <span> Prescription Use (Part 21 CFR 801 Subpart D) </span> <span> Over-The-Counter Use (21 CFR 801 Subpart C) </span> |

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# 1. 510(k) Summary

## A. Device Information

| Category | Comments |
|----------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------|
| Sponsor: | Microgenics Corporation<br>Thermo Fisher Scientific<br>46500 Kato Road<br>Fremont, CA 94538<br>Phone: [phone]<br>FAX: [phone] |
| Correspondent Contact<br>Information: | Sharron Penn<br>Senior Regulatory Affairs Specialist<br>Email: [e-mail]<br>Phone: [phone]<br>FAX: [phone] |
| Device Common Name: | 6-Acetylmorphine Immunoassay Test |
| Trade or Proprietary Name | CEDIA Heroin Metabolite (6-AM) Assay |
| Predicate Device Product<br>Code, Classification,<br>Classification Name & Panel | DJG, Class II, 21 CFR 862.3650 – Opiate test system,<br>91 - Toxicology |

## Predicate Device Information:

| Predicate Device: | CEDIA Heroin Metabolite (6-AM) Assay |
|----------------------------------------------------------------------------------|---------------------------------------------------------------------------|
| Predicate Device<br>Manufacturer: | Microgenics Corporation |
| Predicate Device Common<br>Name: | 6-Acetylmorphine Immunoassay Test System |
| Predicate Device Premarket<br>Notification #: | K173183 |
| Predicate Device Product<br>Code, Classification,<br>Classification Name & Panel | DJG, Class II, 21 CFR 862.3650 ">– Opiate test system,<br>91 – Toxicology |

## B. Date Summary Prepared December 12, 2019

# C. Description of Device

The assay consists of buffers (1 and 2) and lyophilized reagents (1a and 2a). The components include mouse monoclonal antibodies to 6-Acetylmorphine, recombinant microbial "enzyme donor" - 6-Acetylmorphine conjugate, "enzyme acceptor", chlorophenol red ß-Dgalactopyranoside, stabilizers and preservatives.

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## D. Intended Use

## CEDIA Heroin Metabolite (6-AM) Assay

The CEDIA Heroin Metabolite (6-AM) Assay is a homogeneous enzyme immunoassay for the in vitro qualitative and/or semi-quantitative determination of the presence of heroin metabolite (6-AM) in human urine at a cut-off concentration of 10 ng/mL. The assay is intended to be used in laboratories and provides a rapid analytical screening procedure to detect 6-Acetylmorphine in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This product is intended to be used by trained professionals only.

The semi-quantitative mode is for the purpose of enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as Liquid Chromatography/tandem mass spectrometry (LC-MS/MS) or permitting laboratories to establish quality control procedures.

## The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. Gas chromatography/ mass spectrometry (GC/MS) or Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.1

Clinical and professional judgment should be applied to any drug of abuse test result, particularly when preliminary results are used. For In Vitro Diagnostic Use Only.

| Characteristic | Predicate Device: CEDIA Heroin Metabolite<br>(6-AM) Assay. K173183. | CEDIA Heroin<br>Metabolite (6-AM)<br>Assay |
|----------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------|
| Intended Use | The CEDIA Heroin Metabolite (6-AM) Assay is a<br>homogeneous enzyme immunoassay for the in vitro<br>qualitative and/or semi-quantitative determination<br>of the presence of heroin metabolite (6-AM) in<br>human urine at a cut-off concentration of 10 ng/mL.<br>The assay is intended to be used in laboratories and<br>provides a rapid analytical screening procedure to<br>detect 6-Acetylmorphine in human urine. The assay<br>is designed for use with a number of clinical<br>chemistry analyzers. This product is intended to be<br>used by trained professionals only.<br>The semi-quantitative mode is for the purpose of<br>enabling laboratories to determine an appropriate<br>dilution of the specimen for confirmation by a<br>confirmatory method such as Liquid<br>Chromatography/tandem mass spectrometry (LC-<br>MS/MS) or permitting laboratories to establish<br>quality control procedures. | same |

## E. Comparison to Predicate Device

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| Characteristic | Predicate Device: CEDIA Heroin Metabolite<br>(6-AM) Assay. K173183. | CEDIA Heroin<br>Metabolite (6-AM)<br>Assay |
|-------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------|
| | The assay provides only a preliminary analytical<br>test result. A more specific alternative chemical<br>method must be used to obtain a confirmed<br>analytical result. Gas chromatography/ mass<br>spectrometry (GC/MS) or Liquid chromatography<br>with tandem mass spectrometry (LC-MS/MS) is<br>the preferred confirmatory method<br><br>Clinical and professional judgment should be<br>applied to any drug of abuse test result, particularly<br>when preliminary results are used. For In Vitro<br>Diagnostic Use Only. | |
| Operating Principle<br>(Technology) | CEDIA | Same |
| Measured Analyte | Heroin and 6-Acetylmorphine | Same |
| Test Matrix | Urine | Same |
| Cutoff Levels | 10 ng/mL | Same |
| Methodology | Homogeneous Enzyme Immunoassay | Same |
| Reagents Form | EA and ED: Lyophilized (Reconstitution<br>Required)<br>EARB and EDRB liquid ready-to-use. | Same |
| Antibody | Mouse Monoclonal antibody | Same |
| Storage | 2-8°C until expiration date. | Same |
| Principal Operator | Trained professionals | Same |
| Reference Instrument | Beckman AU680 | Horiba Pentra C400 |

## F. Test Principle

The CEDIA Heroin Metabolite Assay uses recombinant DNA technology (US Patent No. 4708929) to produce a unique homogeneous enzyme immunoassay system.10 This assay is based on the bacterial enzyme ß-galactosidase, which has been genetically engineered into two inactive fragments. These fragments, termed Enzyme Acceptor (EA) and Enzyme Donor (ED) spontaneously re-associate to form a fully active enzyme that, in the assay format, cleaves a substrate, generating a color change that can be measured spectrophotometrically.

# G. Summary of Supporting Data

# 1. Analytical Performance:

- a) Precision Study is performed for two replicates per run, twice a day, for 20 days (total n=80). Samples are prepared by spiking 6-Acetylmorphine methanol stock solution into drug free urine at the cutoff, 25%, 50%, 75% & 100% above and below the cutoff and tested in both qualitative and semi-quantitative modes, using a Clinical Laboratory and Standards Institute (CLSI) protocol.

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#### Qualitative Results:

| 6-<br>Acetylmorphine | % of Cutoff<br>(10 ng/mL) | LC-MS/MS<br>(ng/mL) | Total Precision (n=80) | |
|------------------------------------|---------------------------|---------------------|-----------------------------|-----------------------------------------------|
| Spiked<br>Concentration<br>(ng/mL) | | | Number of<br>Determinations | Immunoassay<br>Results<br>(Negative/Positive) |
| 0 | -100 | N/A | 80 | 80/0 |
| 2.5 | -75 | 2.6 | 80 | 80/0 |
| 5 | -50 | 5.1 | 80 | 80/0 |
| 7.5 | -25 | 7.6 | 80 | 80/0 |
| 10 | 100 | 10.0 | 80 | 25/55 |
| 12.5 | +25 | 12.6 | 80 | 0/80 |
| 15 | +50 | 15.0 | 80 | 0/80 |
| 17.5 | +75 | 17.6 | 80 | 0/80 |
| 20 | +100 | 20.0 | 80 | 0/80 |

#### Semi-Quantitative Results:

| 6-<br>Acetylmorphine<br>Spiked<br>Concentration<br>(ng/mL) | % of Cutoff<br>(10 ng/mL) | LC-MS/MS<br>(ng/mL) | Total Precision (n=80) | |
|------------------------------------------------------------|---------------------------|---------------------|-----------------------------|-----------------------------------------------|
| | | | Number of<br>Determinations | Immunoassay<br>Results<br>(Negative/Positive) |
| 0 | -100 | N/A | 80 | 80/0 |
| 2.5 | -75 | 2.6 | 80 | 80/0 |
| 5 | -50 | 5.1 | 80 | 80/0 |
| 7.5 | -25 | 7.6 | 80 | 80/0 |
| 10 | 100 | 10.0 | 80 | 46/34 |
| 12.5 | +25 | 12.6 | 80 | 0/80 |
| 15 | +50 | 15.0 | 80 | 0/80 |
| 17.5 | +75 | 17.6 | 80 | 0/80 |
| 20 | +100 | 20.0 | 80 | 0/80 |

- b) Spike Recovery In qualitative mode, there is no ± 2SD overlap between the spiked 10 ng/mL, 7.5ng/mL and 12.5 ng/mL samples.
All 20 replicates of spiked 7.5 ng/mL and 12.5 ng/mL samples detect as Negative and Positive, respectively, when compared to 10 ng/mL spiked sample.

In semi-quantitative mode, the spiked samples recover within 80-120% of the nominal values.

- c) Analytical Recovery and Dilution Linearity -To demonstrate the dilution linearity for purpose of sample dilution and quality control of the entire assay range, drug free urine is spiked to the high calibrator level (20ng/mL) by using 6-Acetylmorphine methanol solution and diluted with drug free urine to generate 10 intermediate levels.

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Each sample is run in replicates of five in semi-quantitative mode and the average is used to determine percent recovery compared to the expected target value.

| Level | Target<br>Concentration<br>(ng/mL) | Observed<br>Concentration<br>(ng/mL) | Average<br>Recovery (%) | Range of<br>Recovery (%) |
|-------|------------------------------------|--------------------------------------|-------------------------|--------------------------|
| 1 | 0 | 0.39 | N/A | N/A |
| 2 | 2 | 2.31 | 115.6 | 92.0 - 134.0 |
| 3 | 4 | 4.62 | 115.6 | 100.8 - 130.5 |
| 4 | 6 | 6.25 | 104.2 | 97.7 - 113.5 |
| 5 | 8 | 8.04 | 100.5 | 94.6 – 106.5 |
| 6 | 10 | 10.19 | 101.9 | 97.0 – 108.4 |
| 7 | 12 | 12.33 | 102.7 | 99.6 – 105.1 |
| 8 | 14 | 14.36 | 102.6 | 98.3 – 106.1 |
| 9 | 16 | 15.81 | 98.8 | 95.4 - 100.1 |
| 10 | 18 | 18.27 | 101.5 | 98.1 - 103.3 |
| 11 | 20 | 19.46 | 97.3 | 95.5 - 99.0 |

- d) Method Comparison and Accuracy One hundred and three patient samples were analyzed by the CEDIA Heroin Metabolite (6-AM) Assay in both qualitative and semiquantitative modes and the results were compared to LC-MS/MS. The overall concordance between LC-MS/MS and CEDIA Heroin Metabolite (6-Acetylmorphine) Assay was 100% for the samples evaluated in this study.
## Qualitative Results

| Candidate<br>Device<br>Results | Negative | < 50% of<br>Cutoff<br>concentration<br>n by LC-<br>MS/MS<br>(< 5ng/mL) | Near Cutoff<br>Negative<br>(Between 50%<br>below the cutoff<br>and the cutoff<br>concentration as<br>determined by<br>LC-MS/MS)<br>(5 – 9.9 ng/mL) | Near Cutoff<br>Positive<br>(Between the<br>cutoff and 50%<br>above the<br>cutoff<br>concentration<br>as determined<br>by LC-MS/MS)<br>(10 – 15.0<br>ng/mL) | High Positives<br>(Greater than<br>50% above<br>cutoff<br>concentration<br>(> 15.0<br>ng/mL) |
|--------------------------------|----------|------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 0 | 6 | 45 |
| Negative | 44 | 2 | 6 | 0 | 0 |

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## Semi-Quantitative Results

| Candidate<br>Device<br>Results | Negative | < 50% of<br>Cutoff<br>concentration<br>n by LC-<br>MS/MS<br>(< 5ng/mL) | Near Cutoff<br>Negative<br>(Between 50%<br>below the cutoff<br>and the cutoff<br>concentration as<br>determined by<br>LC-MS/MS)<br>(5 – 9.9 ng/mL) | Near Cutoff<br>Positive (Between<br>the cutoff and<br>50% above the<br>cutoff<br>concentration as<br>determined by<br>LC-MS/MS)<br>(10 – 15.0 ng/mL) | High Positives<br>(Greater than<br>50% above<br>cutoff<br>concentration<br>(> 15.0<br>ng/mL) |
|--------------------------------|----------|------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------|
| Positive | 0 | 0 | 0 | 6 | 45 |
| Negative | 44 | 2 | 6 | 0 | 0 |

- e) Specificity The cross-reactivity of Heroin Metabolite (6-AM) and its metabolites was evaluated by adding known amounts of each analyte to drug-free negative urine. As indicated by the results in the table below, 6-Acetylmorphine showed 100% crossreactivity. Heroin showed 8.3% cross-reactivity.

| Heroin Metabolite (6-AM) and its<br>metabolites | Tested Concentration<br>(ng/mL) | Positive/<br>Negative | Cross-<br>reactivity<br>(%) |
|-------------------------------------------------|---------------------------------|-----------------------|-----------------------------|
| 6-Acetylmorphine | 10 | Positive | 100 |
| Heroin | 120 | Positive | 8.3 |

## Cross Reactivity of Structurally Related or Unrelated Opiate Compounds

| Structurally related compounds and<br>other opiates | Tested<br>Concentration<br>(ng/mL) | Positive/<br>Negative | Cross-reactivity (%) |
|-----------------------------------------------------|------------------------------------|-----------------------|----------------------|
| 6-Acetylcodeine | 100,000 | Negative | <0.01 |
| Buprenorphine | 100,000 | Negative | <0.01 |
| Buprenorphine-3ß-D-glucuronide | 100,000 | Negative | <0.01 |
| Codeine | 100,000 | Negative | <0.01 |
| Dextromethorphan | 90,000 | Positive | 0.01 |
| Dihydrocodeine | 100,000 | Negative | <0.01 |
| EDDP | 100,000 | Negative | <0.01 |
| EMDP | 100,000 | Negative | <0.01 |
| Ethylmorphine | 100,000 | Negative | <0.01 |
| Fentanyl | 100,000 | Negative | <0.01 |
| Hydrocodone | 100,000 | Negative | <0.01 |
| Hydromorphone | 20,000 | Positive | 0.05 |
| Hydromorphone-3B-D-glucuronide | 100,000 | Negative | <0.01 |
| LAAM | 100,000 | Negative | <0.01 |
| Levorphanol | 15,000 | Positive | 0.07 |
| Methadone | 100,000 | Negative | <0.01 |
| Meperidine | 100,000 | Negative | <0.01 |
| Mitragynine | 100,000 | Negative | <0.01 |
| 7-Hydroxymitragynine | 100,000 | Negative | <0.01 |
| Morphine | 13,500 | Positive | 0.07 |

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| Structurally related compounds and<br>other opiates | Tested<br>Concentration<br>(ng/mL) | Positive/<br>Negative | Cross-reactivity (%) |
|-----------------------------------------------------|------------------------------------|-----------------------|----------------------|
| Morphine-3β-D-Glucuronide | 100,000 | Negative | <0.01 |
| Morphine-6β-D-Glucuronide | 100,000 | Negative | <0.01 |
| Nalorphine | 10,500 | Positive | 0.1 |
| Naloxone | 100,000 | Negative | <0.01 |
| Naltrexone | 100,000 | Negative | <0.01 |
| Norbuprenorphine | 100,000 | Negative | <0.01 |
| Norbuprenorphine glucuronide | 100,000 | Negative | <0.01 |
| Norcodeine | 100,000 | Negative | <0.01 |
| Norhydrocodone | 100,000 | Negative | <0.01 |
| Normorphine | 50,000 | Positive | 0.02 |
| Norpropoxyphene | 100,000 | Negative | <0.01 |
| Noroxycodone | 100,000 | Negative | <0.01 |
| Noroxymorphone | 100,000 | Negative | <0.01 |
| Oxycodone | 100,000 | Negative | <0.01 |
| Oxymorphone | 100,000 | Negative | <0.01 |
| Oxymorphone-3β-D-glucuronide | 100,000 | Negative | <0.01 |
| Tapentadol HCl | 100,000 | Negative | <0.01 |
| Tramadol | 100,000 | Negative | <0.01 |

The potential cross-reactivity posed by drugs commonly co-administered with Heroin Metabolite (6-AM) was evaluated by adding each substance to Heroin Metabolite (6-AM) spiked at Low Control (7.5 ng/mL) and High Control (12.5 ng/mL) levels at the concentrations indicated. As shown in the table below, the Controls were detected accurately, the Low Control as negative and the High Control as positive, indicating that all the compounds evaluated exhibited minimal cross reactivity at the concentrations tested.

## Structurally Unrelated Compounds Spiked at the Concentration Listed Below into Low Control and High Control

| Cross reactants | Spiked<br>Concentration<br>(ng/mL) | Spiked 6-Acetylmorphine Level | |
|----------------------------|------------------------------------|-------------------------------|--------------|
| | | Low Control | High Control |
| 10,11 Dihydrocarbamazepine | 85,000 | Negative | Positive |
| 11-nor-Δ9-THC-COOH | 10,000 | Negative | Positive |
| Acetaminophen | 500,000 | Negative | Positive |
| Acetylsalicylic Acid | 500,000 | Negative | Positive |
| Amitriptyline | 125,000 | Negative | Positive |
| Amoxicillin | 500,000 | Negative | Positive |
| Amphetamine | 100,000 | Negative | Positive |
| Amisulpride | 100,000 | Negative | Positive |
| Benzotropine Mesylate | 125,000 | Negative | Positive |
| Benzoylecgonine | 100,000 | Negative | Positive |
| Brompheniramine | 75,000 | Negative | Positive |
| Caffeine | 500,000 | Negative | Positive |
| | Spiked | Spiked 6-Acetylmorphine Level | |
| Cross reactants | Concentration<br>(ng/mL) | Low Control | High Control |
| Captopril | 500,000 | Negative | Positive |
| Chlordiazepoxide | 100,000 | Negative | Positive |
| Chlorpromazine | 10,000 | Negative | Positive |
| Clomipramine | 250,000 | Negative | Positive |
| Chloroquine | 500,000 | Negative | Positive |
| Cimetidine | 500,000 | Negative | Positive |
| Desipramine | 125,000 | Negative | Positive |
| Diazepam | 100,000 | Negative | Positive |
| Digoxin | 100,000 | Negative | Positive |
| Diphenhydramine | 50,000 | Negative | Positive |
| Doxepine HCl | 50,000 | Negative | Positive |
| Enalapril | 500,000 | Negative | Positive |
| Fluoxetine | 500,000 | Negative | Positive |
| Fluophenazine | 500,000 | Negative | Positive |
| Haloperidol | 50,000 | Negative | Positive |
| Hydroxychlroquine | 100,000 | Negative | Positive |
| Hydroxyzine | 250,000 | Negative | Positive |
| Ibuprofen | 500,000 | Negative | Positive |
| Imipramine | 50,000 | Negative | Positive |
| Levothyroxine | 50,000 | Negative | Positive |
| Methamphentamine | 100,000 | Negative | Positive |
| Maprotiline | 500,000 | Negative | Positive |
| Nalbuphine | 100,000 | Negative | Positive |
| Naproxen | 500,000 | Negative | Positive |
| Nortriptyline | 250,000 | Negative | Positive |
| Nifedipine | 500,000 | Negative | Positive |
| Nordiazepam | 100,000 | Negative | Positive |
| Oxazepam | 100,000 | Negative | Positive |
| Perphenazine | 150,000 | Negative | Positive |
| Phencyclidine | 7,500 | Negative | Positive |
| Phenobarbital | 100,000 | Negative | Positive |
| Procyclidine | 400,000 | Negative | Positive |
| Propoxyphene | 25,000 | Negative | Positive |
| Protriptyline | 50,000 | Negative | Positive |
| Ranitidine | 500,000 | Negative | Positive |
| Salicyluric Acid | 500,000 | Negative | Positive |
| Secobarbital | 100,000 | Negative | Positive |
| Sulpiride | 500,000 | Negative | Positive |
| Thioridazine | 250,000 | Negative | Positive |
| | Spiked<br>Concentration<br>(ng/mL) | Spiked 6-Acetylmorphine Level | |
| Cross reactants | | Low Control | High Control |
| Triprolidine | 125,000 | Negative | Positive |
| Verapamil | 500,000 | Negative | Positive |

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

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

- Interference The potential interference of pH and endogenous physiologic substances f) on recovery of 6-Acetylmorphine using CEDIA Heroin Metabolite (6-AM) Assay was assessed by spiking known compounds of potentially interfering substances into the Low Control (7.5 ng/mL) and High Control (12.5 ng/mL). In the presence of the compounds listed below, the controls are detected accurately, indicating that these compounds did not show interference in the assay.

| | Tested<br>Concentration<br>(mg/dL) | Spiked 6-Acetylmorphine Level | |
|---------------------|------------------------------------|-------------------------------|--------------|
| Compound | | Low Control | High Control |
| Acetone | 1000 | Negative | Positive |
| Ascorbic acid | 1500 | Negative | Positive |
| Creatinine | 500 | Negative | Positive |
| Ethanol | 1000 | Negative | Positive |
| Galactose | 10 | Negative | Positive |
| Y-globulin | 500 | Negative | Positive |
| Glucose | 1000 | Negative | Positive |
| Hemoglobin | 300 | Negative | Positive |
| Human serum albumin | 500 | Negative | Positive |
| Oxalic acid | 100 | Negative | Positive |
| Riboflavin | 7.5 | Negative | Positive |
| Sodium Chloride | 6000 | Negative | Positive |
| Urea | 2000 | Negative | Positive |
| pH | 3.0 | Negative | Positive |
| pH | 4.0 | Negative | Positive |
| pH | 5.0 | Negative | Positive |
| pH | 6.0 | Negative | Positive |
| pH | 7.0 | Negative | Positive |
| pH | 8.0 | Negative | Positive |
| pH | 9.0 | Negative | Positive |
| pH | 10.0 | Negative | Positive |
| pH | 11.0 | Negative | Positive |

Specific Gravity - Drug free urine samples with specific gravity ranging in value within 1.002 to 1.029 were split and spiked to a final concentration of either 7.5 ng/mL or 12.5 ng/mL (the Low Control and High Control concentrations, respectively). No interference is observed.

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

| | Spiked 6-Acetylmorphine Concentration | |
|------------------|---------------------------------------|--------------|
| Specific Gravity | Low Control | High Control |
| 1.002 | Negative | Positive |
| 1.004 | Negative | Positive |
| 1.005 | Negative | Positive |
| 1.007 | Negative | Positive |
| 1.010 | Negative | Positive |
| 1.012 | Negative | Positive |
| 1.014 | Negative | Positive |
| 1.019 | Negative | Positive |
| 1.023 | Negative | Positive |
| 1.025 | Negative | Positive |
| 1.029 | Negative | Positive |

## g) Stability

## Open Vial Stability:

Open Vial stability studies for one lot stored at 2-8°C supports the claim of 60 days for qualitative and semi-quantitative modes. This data was presented in the original 510(k) submission K173183.

## Reagent On-Board Stability:

Reagent On-Board stability studies for one lot stored on-board clinical analyzer supports the claim of 60 days for qualitative and semi-quantitative modes.

#### Reconstituted Reagent Stability:

Reconstituted Reagent stability studies for one lot stored at 2-8°C supports the claim of 60 days for qualitative and semi-quantitative modes.

## Real Time Stability for Reagent

Real time stability studies for three lots of reagents stored at 2-8°C have been carried out for up to 26 months. Proposed shelf-life claim is 24 months. This data was presented in the original 510(k) submission K173183.

- h) Detection Limit Limit of detection is not provided in a 510(k) of this type of assay.
- Clinical Sensitivity Clinical sensitivity is not provided in a 510(k) of this type of assay. i)
- Clinical Specificity Clinical specificity is not provided in a 510(k) of this type of assay. i)
- k) Clinical cutoff SAMHSA guideline for 6-Acetylmorphine is 10 ng/mL cutoff.

#### H. Conclusion

The information supports a determination of substantial equivalence between CEDIA Heroin Metabolite (6-AM) Assay and the predicate device CEDIA Heroin Metabolite (6-AM) Assay (K173183)