CEDIA BUPRENORPHINE ASSAY

K040316 · Microgenics Corp. · DJG · Jun 23, 2004 · Clinical Toxicology

Device Facts

• Record ID: K040316
• Device Name: CEDIA BUPRENORPHINE ASSAY
• Applicant: Microgenics Corp.
• Product Code: DJG · Clinical Toxicology
• Decision Date: Jun 23, 2004
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.3650
• Device Class: Class 2

Indications for Use

The CEDIA® Buprenorphine assay is a homogenous enzyme immunoassay for qualitative or semi-quantitative determination of the presence of buprenorphine in human urine at cutoff concentration of 5 ng/mL. The assay provides a simple and rapid analytical screening procedure to detect buprenorphine in human urine. The CEDIA® Buprenorphine calibrators are used to calibrate the CEDIA® Buprenorphine Assay in human urine. The CEDIA® Buprenorphine controls are used to qualify the CEDIA® Buprenorphine Assay in human urine. 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) 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.

Device Story

Homogeneous enzyme immunoassay for buprenorphine detection in human urine; utilizes automated clinical chemistry analyzers. Principle: bacterial B-galactosidase engineered into two inactive fragments; one fragment conjugated to buprenorphine. Sample buprenorphine competes with conjugate for anti-buprenorphine antibody binding; free conjugate allows enzyme fragment reassociation. Reassociated enzyme cleaves substrate, producing color change measured spectrophotometrically at 660 nm. Output: qualitative or semi-quantitative concentration. Used in clinical laboratory settings by technicians. Results serve as preliminary screening; require GCMS confirmation for clinical decision-making. Benefits: rapid identification of buprenorphine presence.

Clinical Evidence

No clinical data. Bench testing only.

Technological Characteristics

Homogenous enzyme immunoassay; recombinant DNA technology (US Patent 4706929) producing β-galactosidase fragments. Spectrophotometric detection. Liquid calibrators and controls. Storage 2°C to 8°C. Designed for use on automated clinical chemistry analyzers.

Indications for Use

Indicated for the qualitative or semi-quantitative screening of buprenorphine in human urine. Provides preliminary analytical results; requires confirmation by alternative chemical method (e.g., GCMS). Does not distinguish between therapeutic use and abuse.

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 DAU 6-Acetylmorphine Assay (K001178)

Submission Summary (Full Text)

{0}

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

A. 510(k) Number:
K040316

B. Purpose for Submission:
New analyte

C. Analyte:
Buprenorphine

D. Type of Test:
Homogeneous enzyme immunoassay, qualitative or semi-quantitative.

E. Applicant:
Microgenics Corporation

F. Proprietary and Established Names:
CEDIA Buprenorphine Assay

G. Regulatory Information:
1. Regulation section:
21CFR862.3650, 21CFR862.3200, 21CFR862.3280
2. Classification:
Class II
3. Product Code:
DJG, DLJ, LAS
4. Panel:
91

H. Intended Use:
1. Intended use(s):
The CEDIA® Buprenorphine assay is a homogeneous enzyme immunoassay for qualitative or semi-quantitative determination of the presence of at a cutoff concentration of 5 ng/ml. The assay provides a simple and rapid analytical screening procedure to detect buprenorphine in human urine.

The CEDIA Buprenorphine calibrators are used to calibrate the CEDIA Buprenorphine Assay in human urine.

The CEDIA Buprenorphine controls are used to qualify the CEDIA Buprenorphine Assay in human urine.

{1}

Page 2 of 6

2. **Indication(s) for use:**
The assay provides a simple and rapid analytical screening procedure to detect buprenorphine in human urine.

3. **Special condition for use statement(s):**
The assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used to obtain a confirmed analytical result. GCMS is the preferred confirmatory method. Clinical and professional judgement should be applied to any drug of abuse test result, particularly when preliminary results are used.

The assay does not distinguish whether buprenorphine is being taken therapeutically or abused.

4. **Special instrument Requirements:**
The assay is for use on automated clinical chemistry analyzers.

I. **Device Description:**
The assay consists of buffers (1 and 2) and lyophilized reagents (1a and 2a). The components include mouse monoclonal anti-buprenorphine antibody, recombinant microbial "enzyme donor" – buprenorphine conjugate, "enzyme acceptor", chlorophenol red B-D-galactopyranoside, stabilizers and preservatives. Calibrators and controls are sold separately.

J. **Substantial Equivalence Information:**
1. **Predicate device name(s):**
CEDIA 6-Acetylmorphine Assay
2. **Predicate K number(s):**
K001178
3. **Comparison with predicate:**
Both devices use similar reagent and instrument systems. Both devices detect opioid derivatives. The specific antibodies used in the devices and the specific compounds detected are different for the two devices. The predicate detects 6-acetylmorphine with a cutoff concentration of 10 ng/ml. This device detects buprenorphine with a cutoff concentration of 5 ng/ml.

K. **Standard/Guidance Document Referenced (if applicable): None**

L. **Test Principle:**
The assay is based on bacterial enzyme B-galactosidase genetically engineered to 2 inactive fragments, one of which is conjugated to buprenorphine. Buprenorphine in the sample competes with the enzyme fragment-conjugated buprenorphine for binding to anti-buprenorphine antibody. In the absence of buprenorphine in the sample, the fragment binds antibody and does not reassociate to form active enzyme. If buprenorphine is present in the sample it binds to the antibody, allowing the enzyme fragments to reassociate. The reassociated enzyme cleaves the substrate,

{2}

Page 3 of 6

generating a color change that can be measured spectrophotometrically (660 nm). The amount of active enzyme is proportional to the analyte present.

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

### 1. Analytical performance:

Performance was evaluated at the manufacturer's site on a Hitachi 717 Analyzer.

### a. Precision/Reproducibility:

Reproducibility was determined by assaying 6 replicates of each of 3 controls in 20 separate runs. Reproducibility was expressed as the %CV for the average concentration of 20 runs for each of the 3 control levels. Results of the inter-assay reproducibility evaluation are shown below.

| | Low control | Mid control | High control |
| --- | --- | --- | --- |
| Total number of samples | 120 | 120 | 120 |
| Average concentration (ng/ml) | 4.4 | 6.8 | 36.5 |
| SD (ng/ml) | 0.2 | 0.3 | 1.4 |
| %CV | 5.0 | 3.8 | 4.0 |

### b. Linearity/assay reportable range:

To evaluate linearity a patient urine pool containing 76.6 ng/ml buprenorphine (as measured using the CEDIA buprenorphine assay) was sequentially diluted using a human urine pool free of buprenorphine. The resulting 10 dilutions were assayed in duplicate within one run. Within the range of 8-76.6 ng/ml, the expected/observed values ranged from 95.5%-107.4%.

### c. Traceability (controls, calibrators, or method):

Calibrator levels are 0, 5, 20, 50, 75 ng/ml. Control levels are approximately 3-4 ng/ml and 6-7 ng/ml i.e. approx 25% above and below the cutoff concentration.

Calibrator stability for opened calibrators is evaluated at 2-8 degrees C for up to 60 days. Closed calibrator stability is determined for calibrators stored at 25 degrees C. Acceptance criteria for recovery, with the CEDIA assay, are +/-10% of the concentration observed on day 0. Real time stability studies are ongoing.

{3}

Page 4 of 6

Calibrators and controls are traceable to a USP drug standard. Master calibrators are gravimetrically prepared and values are confirmed by GCMS analysis.

d. Detection limit:
The limit of blank of the assay is based on the average plus 3 standard deviations of measurements of 21 buprenorphine-free specimens and was evaluated on 3 instruments. The limit of the blank was 1.25 ng/ml.

e. Analytical specificity
To evaluate interference by endogenous compounds, known amounts of potentially interfering substances were added to urine specimens spiked with 5 ng/ml buprenorphine. Samples containing each endogenous substance and a matched control were assayed using the CEDIA Buprenorphine Assay. The substances tested and percent recoveries are listed below.

| Substance tested | Concentration (mg/dl) | % recovery (relative to control) |
| --- | --- | --- |
| Acetone | 1000 | 98.1 |
| Ascorbic acid | 1500 | 91.2 |
| Creatinine | 500 | 101.8 |
| Galactose | 10 | 108.2 |
| Gamma globulin | 500 | 93.4 |
| Glucose | 1500 | 93.0 |
| Hemoglobin | 300 | 101.2 |
| NaCl | 6000 | 100.0 |
| Oxalic Acid | 100 | 103.0 |
| Human serum albumin | 500 | 97.2 |
| Urea | 2000 | 93.5 |
| Riboflavin | 7.5 | 91.7 |
| Ethanol | 1000 | 108.0 |

Cross-reactivity of buprenorphine metabolites were determined by adding known amounts of metabolites to buprenorphine-free urine specimens. Concentrations were determined and compared to control samples free of metabolites. Results are tabulated below:

| Metabolite | Target concentration (ng/ml) | % cross-reactivity |
| --- | --- | --- |
| Buprenorphine 3-B-D | 5 | 98 |
| Glucuronide | 20 | 97 |
| Norbuprenorphine | 1000 | <0.01 |
| Norbuprenorphine 3-B-D | 1000 | <0.01 |
| Glucuronide | | |

{4}

Potential interference caused by other substances or drugs was evaluated by adding 500 ug/ml of each drug to buprenorphine-free urine and comparing results to those of controls samples. Percent cross-reactivity observed at 500 ug/ml was less than 0.1% for the compounds listed below.

| Pharmacologic Compound | |
| --- | --- |
| 10, 11 Dihydrocarbamazepine
11-nor-THC-COOH
Acetaminophen
Acetylsalicyclic Acid
Amitriptyline
Amoxicillin
Amphetamine
Benzotropine methane
Sulfonate
Benzoylecgonine
Bromopheniramine
Caffeine
Captopril
Chlordiazepoxide
Chlorpromazine
Cimetidine
Desipramone
Diazepam
Digoxin
Diphenhydramine
Disopyramide
Doxepin
Doxylamine | Enalapril
Fluoxetine
Haloperidol
Ibuprofen
Levothyroxine
Methamphetamine
Nifedipine
Nordiazepam
Oxazepam
Pentazocine HCL
Perphenazine
Phencyclidine
Phenobarbital
Procyclidine
Propoxyphene
Protriptyline
Ranitidine
Salicyluric Acid
Secobarbital
Tolmetin
Triprolidine
Verapamil |

f. Assay cut-off:

Urine specimens (n=21) were spiked with a stock solutions of buprenorphine to final concentrations of 3.75 ng/ml and 6.25 ng/ml and evaluated using the CEDIA Buprenorphine Assay. In both the quantitative and qualitative modes, all specimens spiked to 3.75 ng/ml were determined by the assay to be below the cutoff calibrator for that run. Similarly, all specimens spiked to 6.25 ng/ml were determined to be above the cutoff calibrator. This is summarized in the table below:

| Mean dose (ng/ml) | 3.7 | 6.6 |
| --- | --- | --- |
| SD (ng/ml) | 0.2 | 0.2 |
| %cv | 6.6 | 2.5 |
| Cut-off dose (ng/ml) | 5.4 | 5.4 |

{5}

Page 6 of 6

2. Comparison studies:

a. Method comparison with predicate device:

Ninety six clinical urine specimens were obtained from normal volunteers and individuals receiving buprenorphine. No selection criteria or pre-screening was applied. Samples spanned the range from 0-75 ng/ml buprenorphine. Results, compared to GCMS analysis for buprenorphine (including buprenorphine glucuronide), are shown below. They were identical for qualitative and semi-quantitative mode. The discordant observation had a GC/MS buprenorphine concentration of 0 ng/mL, a negative result for qualitative mode, and a reported result of 8.3 ng/mL for semi-quantitative mode.

| | GC/MS Positive | GC/MS Negative | |
| --- | --- | --- | --- |
| CEDIA Positive | 45 | 1 | 46 |
| CEDIA Negative | 0 | 50 | 50 |
| | 45 | 51 | 96 |

b. Matrix comparison:

Not applicable. Urine is the only matrix for which the assay is indicated

3. Clinical studies:

a. Clinical sensitivity:

Not applicable. Clinical sensitivity is not typically provided in a 510(k) for this type of assay.

b. Clinical specificity:

Not applicable. Clinical sensitivity is not typically provided in a 510(k) for this type of assay.

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

4. Clinical cut-off:

There are currently no SAMHSA recommendations for a clinical cutoff for buprenorphine.

5. Expected values/Reference range:

NA

N. Conclusion:

The submitted material in this premarket notification is complete and supports a substantial equivalence decision.