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

Indications for 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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K192943 - Page 1 of 12

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

ASSAY ONLY

## I Background Information:

A 510(k) Number

K192943

B Applicant

Microgenics Corporation

C Proprietary and Established Names

CEDIA Heroin Metabolite (6-AM) assay

D Regulatory Information

| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| DJG | Class II | 21 CFR 862.3650 - Opiate Test System | TX - Clinical Toxicology |

## II Submission/Device Overview:

A Purpose for Submission:

Clearance of the CEDIA Heroin Metabolite (6-AM) assay on the Horiba Pentra C400 clinical chemistry analyzer.

B Measurand:

Heroin Metabolite, 6-Acetylmorphine (6-AM).

C Type of Test:

Qualitative and Semi-quantitative Homogeneous Enzyme Immunoassay

## III Intended Use/Indications for Use:

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# A Intended Use(s):

See Indications for Use below.

# B Indication(s) for 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.

# C Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

For in vitro diagnostic use only.

# D Special Instrument Requirements:

Performance characteristics studies were conducted on the Horiba Pentra C400 clinical chemistry analyzer.

# IV Device/System Characteristics:

# A Device Description:

CEDIA Heroin Metabolite (6-AM) Assay is supplied as a two liquid buffers (1 and 2) and two lyophilized reagents (1a and 1b) kit that is available in a kit configuration of 3x17mL.

Kit Constituents:

1 EA Reconstitution Buffer: Contains 0.32 mg/L mouse monoclonal antibodies to 6-Acetylmorphine, buffer salts, stabilizer, detergent and preservative

1a EA Reagent: Contains 0.171 g/L Enzyme Acceptor, buffer salts, detergent and preservative.

2 ED Reconstitution Buffer: Contains buffer salts, stabilizer, and preservative

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2a ED Reagent: Contains 16.2 µg/L Enzyme Donor conjugated to 6-Acetylmorphine, 1.67 g/L chlorophenol red-β-D galactopyranoside, stabilizer, detergent, preservative.

## B Principle of Operation:

CEDIA technology uses recombinant DNA technology to produce a homogeneous enzyme immunoassay system. The assay is based on the bacterial enzyme β-galactosidase, which has been genetically engineered into two inactive fragments. These fragments spontaneously re-associate to form fully active enzymes that, in the assay format, cleave a substrate. This generates a color change that can be measured spectrophotometrically.

In the CEDIA Heroin Metabolite (6-AM) Assay, the analyte in the sample competes with 6-AM conjugated to Enzyme Donor (ED) for antibody binding sites. If 6-AM is present in the sample, it binds to antibody, leaving the ED-6-AM conjugate free to re-associate with Enzyme Acceptor (EA) to form active β-galactosidase. If no 6-AM is present in the sample, antibody binds to the ED-6-AM conjugate, inhibiting the re-association of inactive -galactosidase fragments, and thus reducing the amount of active enzyme formed. The amount of active enzyme formed, and resultant absorbance change are proportional to the amount of 6-AM present in the sample.

## V Substantial Equivalence Information:

### A Predicate Device Name(s):

CEDIA Heroin Metabolite (6-AM) Assay

### B Predicate 510(k) Number(s):

K173183

### C Comparison with Predicate(s):

| Device & Predicate Device(s): | K192943 | K173183 |
| --- | --- | --- |
| Device Trade Name | CEDIA Heroin Metabolite (6-AM) Assay | CEDIA Heroin Metabolite (6-AM) Assay |
| General Device Characteristic Similarities | | |
| Intended Use/Indications For Use | 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. | Same |

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| Operating Principle (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 Required) 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 |
| General Device Characteristic Differences | | |
| Instrument used for performance characteristics studies | Horiba Pentra C400 | Beckman AU680 |

VI Standards/Guidance Documents Referenced:

- CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition; 2014.
- CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; 2003.
- CLSI EP07-A2: Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition; 2005.
- CLSI EP25-A: Evaluation of Stability in In Vitro Diagnostic Reagents; Approved Guideline; 2009.

VII Performance Characteristics (if/when applicable):

A Analytical Performance:

All analytical performance studies were conducted on the Horiba Pentra C400 clinical chemistry analyzer.

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1. Precision/Reproducibility:

Precision studies were performed in accordance with CLSI Guideline EP05-A3 at one site with one analyzer and two lots of reagents. The study was performed for 20 days with two runs per day, at least two hours apart and two replicates per run in both Qualitative and Semi-quantitative modes, giving a total of 80 determinants (n = 80). Drug-free negative urine was spiked with 6-Acetylmorphine analyte to final concentrations of -100%, -75%, -50%, -25%, below cutoff, at cutoff and +25%, +50%, +75% and +100%, above cutoff. The concentration of the spiked samples was confirmed by LC-MS/MS or GC/MS. The results from a representative lot are summarized in the tables below.

Qualitative Results:

| 6-Acetylmorphine Concentration (ng/mL) | % of Cutoff (10 ng/mL) | Total Precision (n=80) | |
| --- | --- | --- | --- |
| | | Number of Determinations | Immunoassay Results (Negative/Positive) |
| 0 | -100 | 80 | 80/0 |
| 2.5 | -75 | 80 | 80/0 |
| 5 | -50 | 80 | 80/0 |
| 7.5 | -25 | 80 | 80/0 |
| 10 | 100 | 80 | 25/55 |
| 12.5 | +25 | 80 | 0/80 |
| 15 | +50 | 80 | 0/80 |
| 17.5 | +75 | 80 | 0/80 |
| 20 | +100 | 80 | 0/80 |

Semi-Quantitative Results:

| 6-Acetylmorphine Concentration (ng/mL) | % of Cutoff (10 ng/mL) | Total Precision (n=80) | |
| --- | --- | --- | --- |
| | | Number of Determinations | Immunoassay Results (Negative/Positive) |
| 0 | -100 | 80 | 80/0 |
| 2.5 | -75 | 80 | 80/0 |
| 5 | -50 | 80 | 80/0 |
| 7.5 | -25 | 80 | 80/0 |
| 10 | 100 | 80 | 46/34 |
| 12.5 | +25 | 80 | 0/80 |
| 15 | +50 | 80 | 0/80 |
| 17.5 | +75 | 80 | 0/80 |
| 20 | +100 | 80 | 0/80 |

2. Analytical Recovery and Dilution Linearity:

To demonstrate linearity of the assay throughout the calibration range of 0 to 20 ng/mL, a drug free-urine pool spiked with 6-AM at 20 ng/mL was serially diluted with drug free urine to generate ten intermediate levels. Each sample was run in replicates of five in semi-

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quantitative mode and the average was used to determine percent recovery compared to the expected target value. The observed result (y) and the target expected result (x) were compared using the least squares regression method. The regression equation and correlation obtained are:

$$
y = 0.9692x + 0.4923; r^2 = 0.9986
$$

The recovery of the samples from the linear range of the assay (0 ng/mL to 20 ng/mL) prepared by dilution ranged from 97.3% to 115.6%.

| Level | Target Concentration (ng/mL) | Observed Concentration (ng/mL) | Average Recovery (%) | Recovery Range (ng/mL) |
| --- | --- | --- | --- | --- |
| 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 |

3. Analytical Specificity/Interference:

Cross-reactivity:

Cross-reactivity of the structurally related and unrelated compounds with the performance of the candidate device was evaluated by adding known amounts of each compound to drug-free negative urine. The samples were tested in duplicate with the candidate device in both the qualitative and semi-quantitative modes. Percent cross-reactivity, in semi-quantitative mode, is calculated as follows: % Cross-reactivity = (Cutoff concentration / Lowest concentration of cross reactant that gives a positive result) x 100

The results are summarized below:

Cross Reactivity of Structurally Related and Unrelated Opiate Compounds

| Heroin Metabolite (6-AM) and its metabolites | Tested Concentration (ng/mL) | Positive/Negative | Cross-reactivity (%) |
| --- | --- | --- | --- |
| 6-Acetylmorphine | 10 | Positive | 100 |
| Heroin | 120 | Positive | 8.3 |
| Structurally related compounds and other opiates | Tested Concentration (ng/mL) | Positive/Negative | Cross-reactivity (%) |
| --- | --- | --- | --- |
| 6-Acetylcodeine | 100,000 | Negative | <0.01 |
| Buprenorphine | 100,000 | Negative | <0.01 |

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| Structurally related compounds and other opiates | Tested Concentration (ng/mL) | Positive/Negative | Cross-reactivity (%) |
| --- | --- | --- | --- |
| Buprenorphine-3β-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-3β-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 |
| 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 |

Interference with Commonly Used Drugs:

Interference from commonly co-administered drugs with 6-AM was evaluated by spiking these compounds into urine samples containing 6-AM at +/-25% of the cut-off levels (7.5 ng/mL and 12.5 ng/mL). The samples were tested in duplicate with the candidate device in both the qualitative and semi-quantitative modes. The following list of potentially interfering compounds showed no interference at the indicated concentration when tested with the candidate device.

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| Potential Interferents | Spiked Concentration (ng/mL) | Spiked 6-Acetylmorphine Level | |
| --- | --- | --- | --- |
| | | 7.5 ng/mL | 12.5 ng/mL |
| 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 |
| 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 |
| Hydroxychloroquine | 100,000 | Negative | Positive |
| Hydroxyzine | 250,000 | Negative | Positive |
| Ibuprofen | 500,000 | Negative | Positive |
| Imipramine | 50,000 | Negative | Positive |
| Levothyroxine | 50,000 | Negative | Positive |
| Methamphetamine | 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 |

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| Potential Interferents | Spiked Concentration (ng/mL) | Spiked 6-Acetylmorphine Level | |
| --- | --- | --- | --- |
| | | 7.5 ng/mL | 12.5 ng/mL |
| 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 |
| Triprolidine | 125,000 | Negative | Positive |
| Verapamil | 500,000 | Negative | Positive |

Interference with Endogenous Substances:

The potential interference of endogenous physiologic substances on recovery of 6-AM using the candidate device was assessed by spiking these potentially interfering substances into drug free urine spiked with 6-AM at 7.5 ng/mL and 12.5 ng/mL concentrations. The samples were tested in replicates of five with the candidate device in both the qualitative and semi-quantitative modes. In the presence of the compounds listed below, the controls are detected accurately, indicating that these compounds do not interfere with the candidate device.

| Compound | Tested Concentration (mg/dL) | Spiked 6-Acetylmorphine Level | |
| --- | --- | --- | --- |
| | | 7.5 ng/mL | 12.5 ng/mL |
| 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 |

Effect of pH:

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The pH of drug-free urine samples was adjusted to be pH 3, 4, 5, 6, 7, 8, 9, 10 and 11, and the samples were spiked with 6-AM to +/-25% of the cutoff (7.5 ng/mL and 12.5 ng/mL). The samples were tested in replicates of five with the candidate device in both the qualitative and semi-quantitative modes. The results demonstrated that the pH does not affect the performance of the candidate.

| pH | Spiked 6-Acetylmorphine Concentration | |
| --- | --- | --- |
| | 7.5 ng/mL | 12.5 ng/mL |
| 3.0 | Negative | Positive |
| 4.0 | Negative | Positive |
| 5.0 | Negative | Positive |
| 6.0 | Negative | Positive |
| 7.0 | Negative | Positive |
| 8.0 | Negative | Positive |
| 9.0 | Negative | Positive |
| 10.0 | Negative | Positive |
| 11.0 | Negative | Positive |

Effect of Specific Gravity:

The specific gravity of drug-free urine samples was adjusted ranging in value within 1.002 to 1.029. These samples were split and spiked to a final concentration of either 7.5 ng/mL or 12.5 ng/mL of 6-AM (+/-25% of the cutoff). The samples were tested in replicates of five with the candidate device in both the qualitative and semi-quantitative modes. The results demonstrated that specific gravity of the urine in the range 1.002 to 1.029 does not affect the performance of the candidate.

| Specific Gravity | Spiked 6-Acetylmorphine Concentration | |
| --- | --- | --- |
| | 7.5 ng/mL | 12.5 ng/mL |
| 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 |

4. Assay Reportable Range:

Not applicable.

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

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The open reagent vial, reagent on-board, reconstituted reagent and real time reagent stability data was reviewed under the premarket submission k173183.

6. Detection Limit:
Not applicable.

7. Assay Cut-Off:
See section VII.A.1.

B Comparison Studies:

1. Method Comparison with Predicate Device:
A method comparison study was conducted using one lot of the reagents. One hundred and three unaltered clinical samples were analyzed using the candidate device in one replicate in both the qualitative and semi-quantitative modes and the results were compared to LC-MS/MS. The results obtained in the qualitative and semi-quantitative modes are summarized below:

Qualitative Results
| Candidate Device Results | Negative | < 50% of Cutoff concentration by LC-MS/MS (< 5ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration as determined by LC-MS/MS) (5.0 – 9.9 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration as determined by LC-MS/MS) (10 – 15.0 ng/mL) | High Positives (Greater than 50% above cutoff concentration (> 15.0 ng/mL) |
| --- | --- | --- | --- | --- | --- |
| Positive | 0 | 0 | 0 | 6 | 45 |
| Negative | 44 | 2 | 6 | 0 | 0 |

Semi-Quantitative Results
| Candidate Device Results | Negative | < 50% of Cutoff concentration by LC-MS/MS (< 5ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration as determined by LC-MS/MS) (5.0 – 9.9 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration as determined by LC-MS/MS) (10 – 15.0 ng/mL) | High Positives (Greater than 50% above cutoff concentration (> 15.0 ng/mL) |
| --- | --- | --- | --- | --- | --- |
| Positive | 0 | 0 | 0 | 6 | 45 |
| Negative | 44 | 2 | 6 | 0 | 0 |

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2. **Matrix Comparison:**
Not applicable.

**C Clinical Studies:**
1. **Clinical Sensitivity:**
Not applicable.
2. **Clinical Specificity:**
Not applicable.
3. **Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):**
None.

**D Clinical Cut-Off:**
Not applicable.

**E Expected Values/Reference Range:**
Not applicable.

**VIII Proposed Labeling:**
The labeling supports the finding of substantial equivalence for this device.

**IX Conclusion:**
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

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