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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

ASSAY ONLY TEMPLATE

A. 510(k) Number:

k173963

B. Purpose for Submission:

Modification of a previously cleared device to replace the anti-benzodiazepine polyclonal goat antibodies with polyclonal sheep antibodies.

C. Measurand:

Benzodiazepines

D. Type of Test:

Qualitative and semi-quantitative homogeneous immunoassay

E. Applicant:

Microgenics Corporation

F. Proprietary and Established Names:

DRI Benzodiazepine Assay

G. Regulatory Information:

Regulation section Classification Product Code Panel 21CFR 862.3170 Class II JXM Toxicology (91)

H. Intended Use:

1. Intended use(s):

Refer to Indications for Use below

2. Indication(s) for use:

The DRI Benzodiazepine Assay is a homogeneous enzyme immunoassay intended for the qualitative and/or semi- quantitative determination of the presence of benzodiazepines and their metabolites in human urine at a cutoff concentration of 200 ng/mL. The assay is

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intended to be used in laboratories and provides a simple and rapid analytical screening procedure to detect benzodiazepines in human urine. The assay is designed for use with a number of clinical chemistry analyzers. This assay is calibrated against Oxazepam. 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/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.

3. Special conditions for use statement(s):

For prescription use only.

4. Special instrument requirements:

Performance data was obtained using the Beckman AU680 clinical chemistry analyzer.

I. Device Description:

The assay consists of reagents (A and E):

Reagent A: Contains sheep polyclonal anti-benzodiazepine antibodies, glucose-6- phosphate (G6P) and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as a preservative.

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

J. Substantial Equivalence Information:

1. Predicate device name(s):

DRI Benzodiazepine Assay

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

k930529

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3. Comparison with predicate:

Similarities Item Candidate Device

k173963 Predicate Device

k930529 Intended Use The qualitative and/or

semi-quantitative determination of the presence of benzodiazepines and their metabolites in human urine

Same

Measured Analyte Benzodiazepine and its metabolites

Same

Test Matrix Urine Same Calibrator Oxazepam Same Cutoff Levels 200 ng/mL Same Methodology Homogeneous enzyme

immunoassay Same

Storage 2–8°C until expiration Same

Differences Item Candidate Device

k173963 Predicate Device

k930529 Antibody Polyclonal sheep antibody Polyclonal goat antibody

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

· CLSI EP05-A3, Evaluation of Precision Performance of Quantitative Measurement Methods; Third Edition, 2014.

· CLSI EP07-A2: Interference Testing in Clinical Chemistry, Approved Guideline – Second Edition, 2005.

· CLSI EP09-A3; Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline – Third Edition, 2013.

· CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedures, a Statistical Approach; Approved Guideline, 2003.

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

L. Test Principle:

The DRI Benzodiazepine Assay is a homogeneous enzyme immunoassay with liquid ready- to-use reagents. The assay uses a specific antibody which can detect benzodiazepines and their metabolites in urine. The assay is based on the competition of an enzyme glucose- 6-phosphate dehydrogenase (G6PDH) labeled drug and the drug from the urine sample for a

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fixed amount of specific antibody binding sites. In the absence of free drug from the sample, the enzyme-labeled drug is bound by the specific antibody and the enzyme activity is inhibited. This phenomenon creates a relationship between drug concentration in urine and the enzyme activity. The enzyme G6PDH activity is determined spectrophotometrically at 340 nm by measuring its ability to convert nicotinamide adenine dinucleotide (NAD) to NADH.

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Precision was evaluated using CLSI Guideline EP05-A3 as a guideline, at one site with one analyzer, two operators, and two lots each of reagents, calibrators and controls. Testing was carried out 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 oxazepam to final concentrations of -100%, -75%, -50%, -25%, below cutoff and +25%, +50%, +75% and +100%, above cutoff, and the concentrations were confirmed by LC-MS/MS. Results are summarized below:

Qualitative Mode Lot 1:

% of Cutoff (200 ng/mL)

Target concentration

(ng/mL)

Measured Conc. (ng/mL)

# of determinants # Negative /

# Positive

-100 0 N/A 80 80/0 -75 50 56 80 80/0 -50 100 102 80 80/0 -25 150 161.5 80 80/0 Cut-off 200 214 80 16/64 +25 250 255.5 80 0/80 +50 300 299 80 0/80+75 350 348 80 0/80 +100 400 403 80 0/80

Qualitative Mode Lot 2:

% of Cutoff (200 ng/mL)

Target concentration

(ng/mL)

Measured Conc. (ng/mL)

# of determinants # Negative /

# Positive

-100 0 N/A 80 80/0-75 50 56 80 80/0-50 100 102 80 80/0-25 150 161.5 80 80/0 Cut-off 200 214 80 70/10

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% of Cutoff (200 ng/mL)

Target concentration

(ng/mL)

Measured Conc. (ng/mL)

# of determinants # Negative /

# Positive

+25 250 255.5 80 0/80 +50 300 299 80 0/80 +75 350 348 80 0/80 +100 400 403 80 0/80

Semi-Quantitative Mode Lot 1:

% of Cutoff (200 ng/mL)

Target concentration

(ng/mL)

Measured Conc. (ng/mL)

# of determinants # Negative /

# Positive

-100 0 N/A 80 80/0 -75 50 56 80 80/0-50 100 102 80 80/0 -25 150 161.5 80 80/0 Cut-off 200 214 80 27/53+25 250 255.5 80 0/80+50 300 299 80 0/80+75 350 348 80 0/80 +100 400 403 80 0/80

Semi-Quantitative Mode Lot 2:

% of Cutoff (200 ng/mL)

Target concentration

(ng/mL)

Measured Conc. (ng/mL)

# ofdeterminants # Negative /

# Positive

-100 0 N/A 80 80/0-75 50 56 80 80/0-50 100 102 80 80/0 -25 150 161.5 80 80/0Cut-off 200 214 80 79/1+25 250 255.5 80 0/80+50 300 299 80 0/80 +75 350 348 80 0/80+100 400 403 80 0/80

b. Linearity/assay reportable range:

A recovery study was performed using CLSI EP06-A guidelines. Samples were prepared by spiking a drug free urine pool with a high concentration of oxazepam and generating serial dilutions to achieve concentrations ranging from 100 ng/mL to 1000 ng/mL and were analyzed in replicates of 5 in semi-quantitative mode on one Beckman AU680 analyzer. The results are shown below.

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Target concentration

(ng/mL)

Observed concentration (ng/mL)

% Recovery

0 -1.0 N/A 100 104.5 104.5 200 196.2 98.1 300 314.7 104.9 400 455.3 113.8 500 565.2 113.0 600 661.0 110.2 700 764.7 109.2 800 872.1 109.0 900 937.3 104.1 1000 1024.9 102.5

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

Traceability: The primary calibrators are traceable to the oxazepam drug purchased from a commercial source which is established at 98% purity. The concentration of the primary calibrator stocks is confirmed by LC-MS/MS from three independent laboratories.

d. Detection limit:

Not applicable.

e. Analytical specificity:

Cross-Reactivity Of Benzodiazepine Compounds and Metabolites

The cross reactivity of benzodiazepine compounds and their metabolites was evaluated by adding known amounts of each compound to drug-free negative urine. The specificity (cross-reactivity) study was performed using one lot of reagents, calibrators and controls in both qualitative and semi-quantitative modes. Percent cross-reactivity was calculated as (Cut-off concentration / Lowest concentration of cross reactant that gives a positive result) x 100. Results are summarized below:

Structurally related and unrelated compounds

Lowest concentration producing a positive

result (ng/mL)

Cross-reactivity (%)

α-Hydroxyalprazolam 110 182 α-Hydroxytriazolam 140 143

Alprazolam 110 182 7-Aminoclonazepam 2,500 8

7-Aminoflunitrazepam 300 67

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Structurally related and unrelated compounds

Lowest concentration producing a positive

result (ng/mL)

Cross-reactivity (%)

7-Aminonitrazepam 300 67 Bromazepam 170 118

Chlordiazepoxide 700 29 Clobazam 150 133

Clonazepam 210 95 Clorazepate 135 148 Delorazepam 150 133 Demoxepam 220 91

Desalkylflurazepam 130 154 Diazepam 110 182 Estazolam 100 200

Flunitrazepam 120 167 Flurazepam 150 133

2-Hydroxyethylflurazepam 120 167 Lorazepam 700 29

Lorazepam glucuronide 50,000 <0.4 Lormetazepam 275 73 Medazepam 325 62 Midazolam 180 111 Nitrazepam 130 154 Norchlordiazepoxide 800 25 Nordiazepam 110 182 Oxaprozin 125,000 0.16 Oxazepam 200 100

Oxazepam glucuronide 50,000 0.4 Prazepam 200 100 Temazepam 160 125

Temazepam glucuronide 50,000 <0.4 Triazolam 130 154

Interference Testing of Structurally Unrelated Compounds

Interference from structurally unrelated compounds was evaluated by spiking these compounds into urine samples containing near cutoff negative (150 ng/mL) and near cutoff positive (250 ng/mL) concentrations of oxazepam. The compounds listed in the table below did not cause any positive or negative interference at the concentrations shown:

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Structurally Unrelated Compounds

Tested Concentration

(ng/mL) 6-Acetyl morphine 100,000

10,11 Dihydrocarbamazepine 100,000 11-nor-Δ9-THC-COOH 100,000

Acetaminophen 1,000,000 Acetylsalicylic acid 1,000,000

Amitriptyline 100,000 Amoxicillin 100,000

Amphetamine 100,000 Amisulpride 100,000

Benzotropine Mesylate 100,000 Benzoylecgonine 100,000 Brompheniramine 100,000

Buprenorphine 100,000 Caffeine 100,000 Captopril 100,000

Chlorpromazine 100,000 Chloroquine 100,000 Cimetidine 100,000

Clomipramine 100,000 Codeine 100,000

Desipramine 100,000 Dextromethorphan 100,000

Digoxin 100,000 Dihydrocodeine 100,000

Diphenhydramine 500,000 Doxepine HCl 100,000

EDDP 100,000 EMDP 25,000

Enalapril 100,000 Fentanyl 100,000

Fluoxetine 500,000 Fluophenazine 100,000

Haloperidol 100,000 Heroin 100,000

Hydrocodone 100,000 Hydromorphone 100,000

Hydroxychloroquine 100,000 Hydroxyzine 100,000

Ibuprofen 100,000 Imipramine 100,000

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Structurally Unrelated Compounds

Tested Concentration

(ng/mL) LAAM 100,000

Levorphanol 100,000 Levothyroxine 100,000

Maprotiline 100,000 Meperidine 100,000 Methadone 100,000

Methamphentamine 100,000 Morphine 100,000

Morphine-3β-D-glucuronide 100,000 Morphine-6β-D-glucuronide 100,000

Nalbuphine 100,000 Nalorphine 100,000 Naloxone 100,000

Naltrexone 100,000 Naproxen 100,000 Nifedipine 100,000 Norcodeine 100,000

Norhydrocodone 100,000 Norfluoxetine 500,000 Noroxycodone 100,000

Noroxymorphone 100,000 Norpropoxyphene 100,000

Norsertraline 62,500 Nortryptiline 100,000 Oxycodone 100,000

Oxymorphone 100,000 Paroxetine 100,000

Perphenazine 100,000 Phencyclidine 100,000 Phenobarbital 100,000 Procyclidine 100,000

Propoxyphene 100,000 Protriptyline 100,000 Ranitidine 100,000

Secobarbital 100,000 Sertraline 62,500 Sulpiride 100,000

Tapentadol 100,000 Thioridazine 100,000

Tramadol 100,000 Triprolidine 100,000

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Structurally Unrelated Compounds

Tested Concentration

(ng/mL) Verapamil 100,000 Zaleplon 100,000 Zolpidem 100,000 Zopiclone 100,000

Interference Testing of Endogenous Compounds

Potential interference from endogenous compounds was evaluated by spiking these compounds into urine samples containing near cutoff negative (150 ng/mL) and near cutoff positive (250 ng/mL) concentrations of oxazepam. The compounds or conditions listed in the table below did not cause any positive or negative interference, either in the qualitative or semi-quantative modes, at the concentrations shown in the table below:

Compounds Tested Conc. (mg/dL)

Acetone 500 Ascorbic Acid 150 Creatinine 400 Ethanol 1000 Galactose 5 Glucose 1,000 Hemoglobin 150 Human Serum Albumin (HSA) 200 Oxalic acid 50 Riboflavin 3 Sodium Chloride 1,000 Urea 1,000

Interference Testing of Specific Gravity and pH:

Interference from specific gravity and pH was evaluated by adjusting the specific gravity and pH of samples with near cutoff negative (150 ng/mL) and near cutoff positive (250 ng/mL) concentrations of oxazepam. The following specific gravity or pH levels did not cause any positive or negative interference:

Specific gravity of 1.004, 1.005, 1.007, 1.010, 1.011, 1.013, 1.019, 1.023, 1.025, 1.029.

pH of 3, 4, 5, 6, 7, 8, 9, 10, and 11.

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f. Assay cut-off:

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

2. Comparison studies:

A method comparison study was performed in accordance with CLSI Guideline EP09-A3. One hundred and six patient urine samples were analyzed by the DRI Benzodiazepine Assay in both qualitative and semi-quantitative modes and the results were compared to LC-MS/MS. The results were the same for the qualitative and semi-quantitative modes and are summarized below.

Candidate Device Results vs. LC/MS-MS Values

Candidate Device Results

Negative by LC- MS/MS

< 50% of Cutoff

concentration by LC-

MS/MS (< 100ng/mL)

Near Cutoff Negative (Between

50% below the cutoff and

the cutoff concentration as determined

by LC- MS/MS) (100 – 199 ng/mL)

Near Cutoff Positive

(Between the cutoff and 50% above the cutoff

concentration as determined

by LC- MS/MS) (200 – 300 ng/mL)

High Positives

(Greater than 50% above

cutoff concentration

(> 300 ng/mL)

Positive 0 1* 3* 5 45 Negative 48 2 2 0 0

*Discordant samples

Sample ID

Qualitative Semi-

Quantitative

LC-MS/MS

Negative/ Positive

Negative/ Positive

Oxazepam (ng/mL)

7-Amino-clonazepam

(ng/mL)

α-Hydroxy-alprazolam

(ng/mL) CA160606-045 Positive Positive 86.20 3154.6 NA CA160926-057 Positive Positive 175.08 410.7 13.5 CA170605-001 Positive Positive 151.52 560.4 1.43 CA160908-003 Positive Positive 192.87 NA 96.3

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b. Matrix comparison:

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

3. Clinical studies:

a. Clinical Sensitivity:

Not applicable.

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.