DRI Hydrocodone Assay

K173195 · Microgenics Corporation · DJG · Feb 13, 2018 · Clinical Toxicology

Device Facts

• Record ID: K173195
• Device Name: DRI Hydrocodone Assay
• Applicant: Microgenics Corporation
• Product Code: DJG · Clinical Toxicology
• Decision Date: Feb 13, 2018
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3650
• Device Class: Class 2

Indications for Use

The DRI Hydrocodone Assay is a homogeneous enzyme immunoassay for the qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine at a cutoff concentration of 300 ng/mL. The assay is intended to be used in laboratories and provides a simple and rapid analytical screening procedure to detect hydrocodone and its metabolites in human urine. The assay is designed for use with a number of clinical chemistry analyzers. 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/tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.

Device Story

DRI Hydrocodone Assay is a liquid, ready-to-use homogeneous enzyme immunoassay for urine drug screening. Input: human urine sample. Principle: competitive binding between drug in sample and G6PDH-labeled drug for fixed antibody sites; free drug in sample displaces labeled drug, increasing enzyme activity. Output: spectrophotometric measurement at 340 nm (NAD to NADH conversion) proportional to drug concentration. Used in clinical laboratories on chemistry analyzers (e.g., Indiko Plus). Results are preliminary; used by clinicians to identify specimens requiring confirmatory testing via LC-MS/MS or GC/MS. Benefits: rapid screening for hydrocodone/metabolites to support clinical decision-making regarding patient medication adherence or toxicology screening.

Clinical Evidence

No clinical data. Performance established via bench testing, including precision studies (CLSI EP5-A3), linearity (CLSI EP6-A), and analytical specificity/interference (CLSI EP7-A2). Method comparison study performed on 136 urine samples against LC-MS/MS reference method.

Technological Characteristics

Homogeneous enzyme immunoassay. Reagents: mouse monoclonal anti-hydrocodone antibody, G6P, NAD, and G6PDH-labeled hydrocodone derivative. Detection: spectrophotometric at 340 nm. Form factor: liquid, ready-to-use. Connectivity: designed for clinical chemistry analyzers. Shelf-life: 18 months; on-board stability: 60 days.

Indications for Use

Indicated for the qualitative and/or semi-quantitative screening of hydrocodone and its metabolites in human urine at a 300 ng/mL cutoff. For prescription use in laboratory settings. Not for definitive diagnosis; requires confirmation by GC/MS or LC-MS/MS.

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

• DRI Hydrocodone assay (k150502)

Submission Summary (Full Text)

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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE

A. 510(k) Number:
k173195

B. Purpose for Submission:
Adding a previously cleared test (DRI Hydrocodone Assay) to the Indiko Family Analyzer

C. Measurand:
Hydrocodone

D. Type of Test:
Qualitative and semi-quantitative homogeneous enzyme immunoassay

E. Applicant:
Microgenics Corporation

F. Proprietary and Established Names:
DRI Hydrocodone Assay

G. Regulatory Information:
1. Regulation section:
21 CFR 862.3650, Opiate test system
2. Classification:
Class II
3. Product code:
DJG
4. Panel:
Toxicology (91)

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H. Intended Use:

1. Intended use(s):

Refer to Indications for Use below

2. Indication(s) for use:

The DRI Hydrocodone Assay is a homogeneous enzyme immunoassay for the qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine at a cutoff concentration of 300 ng/mL. The assay is intended to be used in laboratories and provides a simple and rapid analytical screening procedure to detect hydrocodone and its metabolites in human urine. The assay is designed for use with a number of clinical chemistry analyzers.

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/tandem mass spectrometry (LC-MS/MS) is the preferred confirmatory method.

3. Special conditions for use statement(s):

For prescription use only.

4. Special instrument requirements:

Performance characteristics provided in this submission were determined on the Indiko Plus Analyzer.

I. Device Description:

The DRI Hydrocodone assay is supplied as a liquid ready-to-use homogeneous enzyme immunoassay, comprised of two reagents (Reagent A and Reagent E) which are bottled separately but sold together within the same kit.

Reagent A: Contains mouse monoclonal anti-Hydrocodone antibody, glucose-6-phosphate (G6P), and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as a preservative.

Reagent E: Contains Hydrocodone derivative labeled with glucose-6-phosphate dehydrogenase (G6PDH) in Tris buffer with sodium azide as a preservative.

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J. Substantial Equivalence Information:

1. Predicate device name(s):
DRI Hydrocodone assay

2. Predicate 510(k) number(s):
k150502

3. Comparison with predicate:
| Similarities and Differences | | |
| --- | --- | --- |
| Item | Candidate Device k173195 | Predicate Device k150502 |
| Intended Use | Homogeneous enzyme immunoassay for the qualitative and/or semi-quantitative determination of the presence of hydrocodone and its metabolites in human urine. | Same |
| Measured Analyte | Hydrocodone and its metabolites | Same |
| Test Matrix | Urine | Same |
| Cutoff Levels | 300 ng/mL | Same |
| Methodology | Homogeneous Enzyme Immunoassay | Same |
| Antibody | Mouse Monoclonal | Same |
| Storage | 2–8°C until expiration date | Same |
| Platform | Indiko Plus | Beckman AU680 |

K. Standard/Guidance Document Referenced (if applicable):

- CLSI EP5-A3: Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline -Third Edition, 2014.
- CLSI EP7-A2: Interference Testing in Clinical Chemistry; Approved Guideline -Second Edition, 2005.
- CLSI EP9-A3: Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline -Third Edition, 2013.
- CLSI EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures, a Statistical Approach; Approved Guideline, 2003.

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- CLSI EP25-A: Evaluation of Stability of Tn Vitro Diagnostic Reagents; Approved Guideline, 2009.

## L. Test Principle:

The DRI Hydrocodone Assay is a homogeneous enzyme immunoassay. The assay uses specific antibodies that can detect Hydrocodone and its metabolites. The assay is based on competition between a drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug from the urine sample, for a fixed amount of specific antibody binding sites. In the absence of free drug from the sample, mouse monoclonal anti-hydrocodone antibody binds to the drug labeled with G6PDH and causes a decrease in enzyme activity. In the presence of free drug, the free drug occupies the antibody binding sites, allowing the drug bound G6PDH to interact with the substrate, resulting in enzyme activity. This phenomenon creates a direct proportional relationship between the drug concentration in urine and enzyme activity. The enzyme activity is determined spectrophotometrically at 340 nm by measuring the conversion of nicotinamide adenine dinucleotide (NAD) to NADH.

## M. Performance Characteristics (if/when applicable):

### 1. Analytical performance:

#### a. Precision/Reproducibility:

A precision study was performed following CLSI EP5-A3. Drug free urine samples were spiked with hydrocodone at 0, 75, 150, 225, 300, 375, 450, 525 and 600 ng/mL, representing 0, 25, 50, 75, 100, 125, 150, 175 and 200% of the device cutoff (300 ng/mL). Each level sample was tested in duplicate per run, two runs per day for twenty consecutive days (total n= 80/level/reagent lot) using 2 lots on one Indiko Plus analyzer in both qualitative and semi-quantitative modes. The results of one representative lot are shown below:

Qualitative Precision Result:

| Concentration as % of the Cutoff Level | Target Hydrocodone spiked concentration (ng/mL) | Hydrocodone concentration determined by LC-MS/MS (ng/mL) | DRI Hydrocodone Assay # Negative / # Positive |
| --- | --- | --- | --- |
| 0 | 0 | 0 | 80 Neg / 0 Pos |
| 25 | 75 | 89.5 | 80 Neg / 0 Pos |
| 50 | 150 | 174.85 | 80 Neg / 0 Pos |
| 75 | 225 | 243.37 | 80 Neg / 0 Pos |
| 100 | 300 | 318.50 | 4 Neg / 76 Pos |
| 125 | 375 | 410.66 | 0 Neg / 80 Pos |
| 150 | 450 | 501.62 | 0 Neg / 80 Pos |
| 175 | 525 | 592.11 | 0 Neg / 80 Pos |
| 200 | 600 | 667.52 | 0 Neg / 80 Pos |

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Semi-Quantitative Precision Result:

| Concentration as % of the Cutoff Level | Target Hydrocodone spiked concentration (ng/mL) | Hydrocodone concentration determined by LC-MS/MS (ng/mL) | DRI Hydrocodone Assay # Negative / # Positive |
| --- | --- | --- | --- |
| 0 | 0 | 0 | 80 Neg / 0 Pos |
| 25 | 75 | 89.5 | 80 Neg / 0 Pos |
| 50 | 150 | 174.85 | 80 Neg / 0 Pos |
| 75 | 225 | 243.37 | 80 Neg / 0 Pos |
| 100 | 300 | 318.50 | 7 Neg / 73 Pos |
| 125 | 375 | 410.66 | 0 Neg / 80 Pos |
| 150 | 450 | 501.62 | 0 Neg / 80 Pos |
| 175 | 525 | 592.11 | 0 Neg / 80 Pos |
| 200 | 600 | 667.52 | 0 Neg / 80 Pos |

b. Linearity/assay reportable range:

A drug recovery study in the semi-quantitative mode was conducted by spiking a drug free urine pool with hydrocodone and using serial dilutions with a negative urine pool to achieve concentrations ranging from 100 ng/mL to 1000 ng/mL, and testing each level on one lot of reagent, calibrators and controls. Each concentration was tested in five replicates on the Indiko Plus analyzer. The mean of the five replicates tested was defined as the observed drug concentration value. The percent recovery range was defined as the range in observed percent recoveries amongst the five replicates at each concentration. The results of are summarized below:

| Target concentration (ng/mL) | Observed concentration (ng/mL) | % Recovery Range |
| --- | --- | --- |
| 0 | 0 | N/A |
| 100 | 95 | 80-120 |
| 200 | 219 | 160-240 |
| 300 | 331 | 240-360 |
| 400 | 427 | 320-480 |
| 500 | 529 | 400-600 |
| 600 | 624 | 480-720 |
| 700 | 696 | 560-840 |
| 800 | 798 | 640-960 |
| 900 | 893 | 720-1080 |
| 1000 | 911 | 800-1200 |

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c. Traceability, Stability, Expected values (controls, calibrators, or methods):

Calibrators for this device were previously cleared in k150502.

d. Detection limit:

Not applicable.

e. Analytical specificity:

An analytical specificity study to evaluate interference from non-structurally and structurally related compounds was performed in the qualitative and semi-quantitative modes. The study design and results were the same for each mode (qualitative and semi-quantitative modes) and are shown in the tables below.

Cross-Reactivity of Hydrocodone and Its Metabolites

To evaluate potential cross-reactivity for the DRI Hydrocodone assay, structurally similar compounds were spiked into drug free urine at concentrations that will yield a result that is equivalent to the 300ng/mL cutoff. Percent cross-reactivity for compounds with no observed cross-reactivity was calculated using the highest concentration tested. The percent cross-reactivity is presented in the table below.

| Hydrocodone and its metabolites | Tested Concentration (ng/mL) | Pos/neg | % Cross-Reactivity |
| --- | --- | --- | --- |
| Hydrocodone | 300 | Pos | 100 |
| Hydromorphone | 325 | Pos | 92 |
| Hydromorphone-3β-glucuronide | 185 | Pos | 162 |
| Norhydrocodone | 13,000 | Pos | 2 |
| Dihydrocodeine | 12,500 | Pos | 2 |

Cross-Reactivity of Opiates and Structurally Related Compounds

| Compounds | Tested Concentration (ng/mL) | % Cross-Reactivity |
| --- | --- | --- |
| 6-Acetyl Morphine | 100,000 | < 0.3 |
| Buprenorphine | 100,000 | < 0.3 |
| Buprenorphine 3β-D-Glucuronide | 100,000 | < 0.3 |
| Codeine | 150,000 | < 0.2 |
| Dextromethorphan | 250,000 | < 0.2 |
| EDDP | 150,000 | < 0.2 |

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| Fentanyl | 100,000 | < 0.3 |
| --- | --- | --- |
| Heroin | 100,000 | < 0.3 |
| Levorphanol | 22,000 | 1.7 |
| Methadone | 100,000 | <0.3 |
| Meperidine | 100,000 | <0.3 |
| Morphine | 150,000 | <0.2 |
| Morphine -3β-D-Glucuronide | 70,000 | <0.4 |
| Morphine -6 β-D-Glucuronide | 75,000 | <0.4 |
| Nalbuphine | 150,000 | <0.3 |
| Naloxone | 17,000 | 2.0 |
| Naltrexone | 75,000 | <0.3 |
| Norbuprenorphine | 100,000 | <0.3 |
| Norcodeine | 150,000 | <0.2 |
| Normorphine | 150,000 | <0.2 |
| Nor-Oxycodone | 110,000 | 0.3 |
| Oxycodone | 14,000 | 2.5 |
| Oxymorphone 6β-D- Glucuronide | 14,000 | 2.2 |
| Oxymorphone | 14,000 | 2.5 |
| Tapentadol | 100,000 | <0.3 |
| Thebaine | 100,000 | <0.3 |
| Tramadol | 100,000 | <0.3 |

## Structurally Unrelated Compounds

Potential interference from non-structurally related drugs was evaluated in the qualitative and semi-quantitative modes, by spiking these compounds at high concentrations in drug free urine spiked with hydrocodone at $\pm 25\%$ of the cutoff (225 and $375~\mathrm{ng / mL}$). Results obtained with two reagent lots were the same, and results from one representative lot are provided in the table below.

| Compounds | Spiked Concentration (ng/mL) | | |
| --- | --- | --- | --- |
| | | -25% Cutoff (225 ng/mL) | +25% Cutoff (375 ng/mL) |
| Acetaminophen | 500,000 | Neg | Pos |
| Acetylsalicylic acid | 500,000 | Neg | Pos |
| Amitryptyline | 100,000 | Neg | Pos |
| Amoxicillin | 100,000 | Neg | Pos |
| Amphetamine | 1,000,000 | Neg | Pos |
| Benzoylecgonine | 1,000,000 | Neg | Pos |
| Caffeine | 100,000 | Neg | Pos |
| Carbamazepine | 500,000 | Neg | Pos |
| Chlorpromazine | 100,000 | Neg | Pos |
| Clomipramine | 10,000 | Neg | Pos |

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| Cimitidine | 500,000 | Neg | Pos |
| --- | --- | --- | --- |
| Desipramine | 100,000 | Neg | Pos |
| Diphenhydramine | 100,000 | Neg | Pos |
| Doxepine | 100,000 | Neg | Pos |
| Ephedrine | 1,000,000 | Neg | Pos |
| Fluoxethine | 100,000 | Neg | Pos |
| Fluphenazine | 100,000 | Neg | Pos |
| Ibuprofen | 500,000 | Neg | Pos |
| Imipramine | 100,000 | Neg | Pos |
| Maprotiline | 100,000 | Neg | Pos |
| Nortryptiline | 100,000 | Neg | Pos |
| Oxazepam | 250,000 | Neg | Pos |
| Phencyclidine (PCP) | 100,000 | Neg | Pos |
| Phenobarbital | 100,000 | Neg | Pos |
| Ranitidine | 500,000 | Neg | Pos |
| Secobarbital | 100,000 | Neg | Pos |
| Thioridazine | 100,000 | Neg | Pos |

## Endogenous Compounds, pH and Specific Gravity

Potentially interfering substances were spiked into drug free urine spiked with 225 ng/mL or 375 ng/mL of hydrocodone. Samples were tested in replicates of five (n=5) in both qualitative and semi-quantitative modes. Results obtained in both modes were identical, and results are provided in the below table.

| Potential Interfering Substances | Concentration tested (mg/dL) | -25% Cutoff (225ng/mL) | +25% Cutoff (375ng/mL) |
| --- | --- | --- | --- |
| Acetaminophen | 10 | Negative | Positive |
| Acetone | 500 | Negative | Positive |
| Acetylsalicylic Acid | 10 | Negative | Positive |
| Ascorbic Acid | 150 | Negative | Positive |
| Caffeine | 10 | Negative | Positive |
| Creatinine | 400 | Negative | Positive |
| Ethanol | 10 | Negative | Positive |
| Galactose | 5 | Negative | Positive |
| Glucose | 1,000 | Negative | Positive |
| Hemoglobin | 150 | Negative | Positive |
| Human Serum Albumin (HSA) | 200 | Negative | Positive |
| Ibuprofen | 10 | Negative | Positive |
| Oxalic acid | 50 | Negative | Positive |
| Riboflavin | 3 | Negative | Positive |
| Sodium Chloride | 1,000 | Negative | Positive |
| Urea | 1,000 | Negative | Positive |

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pH: Drug free urine specimens spiked with 225 or 375 ng/mL of hydrocodone were adjusted to varying pH (between pH 4.0 and pH 10.0). Samples were tested in replicates of five (n=5) in both qualitative and semi-quantitative modes. No positive or negative interference was observed at urine pH values ranging from 4.0 to 10.0 for each test mode.

Specific Gravity: Urine with specific gravity spanning the range within 1.003–1.031 was spiked with 225 and 375 ng/mL of hydrocodone. Samples were tested in replicates of five (n=5) in both qualitative and semi-quantitative. No positive or negative interference was observed at urine specific gravity values ranging from 1.003 to 1.031 in either test mode.

Assay cut-off:

Characterization of how the device performs analytically around the claimed cutoff concentration of 300 ng/mL hydrocodone is described in the precision section, M.1.a. above.

2. Comparison studies:

a. Method comparison with predicate device:

A total of 136 unaltered urine samples were analyzed by one lot of the DRI Hydrocodone Assay in one replicate, in both Qualitative and Semi-quantitative modes, and the results were compared to LC-MS/MS (liquid- chromatography-tandem mass spectroscopy). The results from the study in the two modes were identical and are summarized in the table below.

| Candidate Device Results | Negative | <50% of cutoff concentration by LC/MS (< 150ng/mL) | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration by LC/MS) (150 ~ 299 ng/mL) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration by LC/MS) (300 ~ 450 ng/mL) | High Positive (Greater than 50% above the cutoff concentration by LC/MS) > 450 ng/mL |
| --- | --- | --- | --- | --- | --- |
| Positive | 0 | 7* | 17* | 9 | 48 |
| Negative | 42 | 3 | 10 | 0 | 0 |

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*Discordant Result Table

| Sample # | Immunoassay result | | |
| --- | --- | --- | --- |
| | | LC-MS/MS (ng/mL) | |
| | | Hydrocodone | Hydromorphone 3β-D Glucuronide |
| 46 | Positive | 126 | 179 |
| 47 | Positive | 132 | 160 |
| 48 | Positive | 126 | 314 |
| 49 | Positive | 82.7 | 128 |
| 50 | Positive | 138 | 677 |
| 51 | Positive | 131 | 126 |
| 52 | Positive | 73.3 | 262 |
| 63 | Positive | 162 | 218 |
| 64 | Positive | 249 | 356 |
| 65 | Positive | 253 | 265 |
| 66 | Positive | 160 | 739 |
| 67 | Positive | 270 | 88.6 |
| 68 | Positive | 217 | 158 |
| 69 | Positive | 236 | 90.4 |
| 70 | Positive | 282 | 242 |
| 71 | Positive | 295 | 433 |
| 72 | Positive | 167 | 149 |
| 73 | Positive | 193 | 756 |
| 74 | Positive | 214 | 434 |
| 75 | Positive | 207 | 706 |
| 76 | Positive | 298 | 190 |
| 77 | Positive | 198 | 262 |
| 78 | Positive | 157 | 124 |
| 79 | Positive | 287 | 79.9 |

b. Matrix comparison:

Not applicable. Urine is the only claimed matrix for the candidate device.

3. Clinical studies:

a. Clinical Sensitivity:

Not applicable.

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b. Clinical specificity:
Not applicable.

c. Other clinical supportive data (when a. and b. are not applicable):
Not applicable.

4. Clinical cut-off:
Not applicable.

5. Expected values/Reference range:
Not applicable.

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Parts 801 and 809, as applicable.

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

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