DRI METHADONE METABOLITE (100/300) ASSAY, CALIBRATORS AND CONTROLS

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k081378 B. Purpose for Submission: New product C. Measurand: 2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) D. Type of Test: Semi-quantitative and qualitative homogeneous enzyme immunoassay E. Applicant: ThermoFisher Scientific F. Proprietary and Established Names: DRI® Methadone Metabolite (100/300) Assay DRI® Methadone Metabolite Urine Calibrators DRI® Methadone Metabolite Urine Controls G. Regulatory Information: 1. Regulation section: 21 CFR §862.3620, Methadone Test System 21 CFR §862.3200, Clinical Toxicology Calibrator 21 CFR §862.3280, Clinical Toxicology Control Material 2. Classification: Class II (Reagent, Calibrator) Class I Reserved (Control) 3. Product code: DJR; DKB; DIF 4. Panel: Toxicology (91) H. Intended Use: 1. Intended use(s): See Indications for Use below. 2. Indication(s) for use: For in vitro diagnostic use only. The DRI Methadone Metabolite (100/300) Assay is intended for the qualitative and semi-quantitative determination of the presence of Methadone Metabolite (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine or EDDP) in human urine at cutoffs of 100 and 300 ng/mL. 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. Tests for methadone metabolite cannot distinguish between abused drugs and certain prescribed medications. Certain foods or medications may interfere with test for {1} methadone metabolite and cause false positive results. The DRI Methadone Metabolite Calibrators are intended for the calibration of the DRI Methadone Metabolite (100/300) Assay. The DRI Methadone Metabolite Controls are intended for use in the DRI Methadone Metabolite (100/300) Assay to detect and monitor systematic deviations from accuracy resulting from reagent or instrument defects. 3. Special conditions for use statement(s): This device is for use by professional laboratory personnel. For in vitro diagnostic use only. 4. Special instrument requirements: Clinical chemistry analyzers. The Hitachi 917 analyzer was used to conduct performance studies below. I. Device Description: The DRI Methadone Metabolite (100/300) Assay utilizes liquid ready-to-use reagents. The Antibody/Substrate Reagent (R1) contains mouse monoclonal anti-EDDP antibody, glucose-6-phosphate (G6P), and nicotinamide adenine dinucleotide (NAD) in Tris buffer with sodium azide as a preservative. The Enzyme Conjugate Reagent (R2) contains EDDP-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): CEDIA DAU EDDP Assay CEDIA DAU Multi-Drug Calibrators MGC DAU Control Sets: Primary, Clinical, Select 2. Predicate 510(k) number(s): K980746; K980853; K040758, 3. Comparison with predicate: | Comparison | DRI Methadone Metabolite (100/300) Assay | Predicate Device – CEDIA DAU EDDP Assay | | --- | --- | --- | | Intended Use | Qualitative and semi-quantitative determination of the presence of Methadone Metabolite (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine or EDDP) in human urine at cutoffs of 100 and 300 ng/mL. | Qualitative and semi-quantitative assay of EDDP (2-ethylidene-1,5-dimethyl-3,3-diphenylpyrrolidine or EDDP) in human urine. | {2} | Comparison | DRI Methadone Metabolite (100/300) Assay | Predicate Device – CEDIA DAU EDDP Assay | | --- | --- | --- | | Test Principle | Homogeneous Enzyme Immunoassay 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. Direct relationship between drug concentration in urine and enzyme activity. Enzyme activity is determined spectrophotometrically at 340 nm by measuring its ability to convert nicotinamide adenine dinucleotide (NAD) to NADH. | Homogeneous Enzyme Immunoassay based on competition between a drug labeled with β-galactosidase, and free drug from the urine sample for a fixed amount of specific antibody binding sites. Direct relationship between drug concentration in urine and enzyme activity. Enzyme activity is determined spectrophotometrically at 570 nm by measuring its ability to convert CPRG to CPR. | | Cutoff | 100 and 300 ng/mL | 100 ng/mL | | Matrix | Human urine | Human urine | | Reagents | Liquid Ready-to-Use Two reagent assay (R1 and R2) | Lyophilized (reconstitution required) Two reagent assay (R1 and R2) | | Calibrators | Liquid ready-to-use (0, 100, 300, 500, 1000 ng/mL) | Liquid ready-to-use (0, 100, 500, 2000 ng/mL) | {3} | Comparison | DRI Methadone Metabolite Control | Predicate Device – MGC Select DAU Control Set | | --- | --- | --- | | Controls | Liquid ready-to-use | Liquid ready-to-use | K. Standard/Guidance Document Referenced (if applicable): CLSI EP5-A: Evaluation of Precision Performance of Quantitative Measurement Methods L. Test Principle: The DRI Methadone Metabolite (100/300) Assay utilizes liquid ready-to-use reagents and calibrators. The assay uses specific antibodies that can detect EDDP in human urine without cross-reactivity to the parent drug methadone. The assay is based on competition between a drug labeled with glucose-6-phosphate dehydrogenase (G6PDH) and free drug from the sample for a fixed number of specific antibody binding sites. In the presence of free drug from the sample, the free drug occupies the antibody binding sites, allowing the drug-labeled G6PDH to interact with the substrate, resulting in enzyme activity. In the absence of free drug from the sample, the specific antibody binds the drug labeled with G6PDH and causes a decrease in enzyme activity. This phenomenon creates a direct relationship between drug concentration in urine and enzyme activity. This enzyme 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: An EDDP solution (1 mg/mL) was added to each of four samples obtained from a human urine sample pool to achieve concentrations that span the assay range. The samples were tested for precision in qualitative and semi-quantitative modes. Following a CLSI (EP5A) precision protocol, samples were tested in 2 replicates per run, 2 runs per day for 20 days, total N=80. Qualitative – 100 cutoff | Drug | Concentration of sample, ng/mL | Number of determinations | Results # Neg / # Pos | | --- | --- | --- | --- | | EDDP | 0 | 80 | 80 Neg / 0 Pos | | EDDP | 50 | 80 | 80 Neg / 0 Pos | | EDDP | 75 | 80 | 80 Neg / 0 Pos | | EDDP | 125 | 80 | 0 Neg / 80 Pos | | EDDP | 225 | 80 | 0 Neg / 80 Pos | Qualitative – 300 cutoff | Drug | Concentration of sample, ng/mL | Number of determinations | Results # Neg / # Pos | | --- | --- | --- | --- | {4} | Drug | Concentration of sample, ng/mL | Number of determinations | Results # Neg / # Pos | | --- | --- | --- | --- | | EDDP | 0 | 80 | 80 Neg / 0 Pos | | EDDP | 125 | 80 | 80 Neg / 0 Pos | | EDDP | 225 | 80 | 80 Neg / 0 Pos | | EDDP | 375 | 80 | 0 Neg / 80 Pos | | EDDP | 750 | 80 | 0 Neg / 80 Pos | Semi-quantitative - 100 cutoff | Drug | Sample Conc., ng/mL | Number of determinations | Result # Neg / # Pos | | --- | --- | --- | --- | | | | | | | EDDP | 0 | 80 | 80/0 | | EDDP | 50 | 80 | 80 / 0 | | EDDP | 75 | 80 | 80 / 0 | | EDDP | 125 | 80 | 0 / 80 | | EDDP | 225 | 80 | 0 / 80 | Semi-quantitative - 300 cutoff | Drug | Sample Conc., ng/mL | Number of determinations | Result # Neg / # Pos | | --- | --- | --- | --- | | | | | | | EDDP | 0 | 80 | | | EDDP | 50 | 80 | 80 / 0 | | EDDP | 75 | 80 | 80 / 0 | | EDDP | 125 | 80 | 80 / 0 | | EDDP | 225 | 80 | 80 / 0 | | EDDP | 375 | 80 | 0 / 80 | | EDDP | 750 | 80 | 0 / 80 | b. Linearity/assay reportable range: The assay linearity was determined by testing the recoveries of a series of samples diluted from a high concentration EDDP sample. A high patient urine sample containing around $1000\mathrm{ng / mL}$ EDDP was serially diluted with analyte-free urine in $10\%$ increments and tested by 5 replicates in semi-quantitative mode. All samples were recovered within $\pm 10\%$ of the expected value and the r-value was 0.9990. c. Traceability, Stability, Expected values (controls, calibrators, or methods): Calibrators and controls are tested on Hitachi 917 analyzers by two runs against a reference (primary) set of calibrators and controls, $N = 5$ for all samples. The acceptance criteria require that replicates have rate imprecision less than $1.5\%$ CV, rate difference between the secondary and the primary calibrators and controls must be within $3\%$ , and concentration of the calibrators and controls must be within $10\%$ error of the nominal values by GC/MS. Calibrators and controls are assigned nominal values if validation {5} results meet acceptance criteria. The assigned values for the calibrators and controls are traceable to the Cerilliant methadone standard catalogue E-022 and are verified by GC/MS. Real time and accelerated stability studies were conducted; protocols and acceptance criteria were described and found to be acceptable. These studies support the manufacturer's stability claims. Real time studies are ongoing. d. Detection limit: Not applicable. This assay is qualitative and semi-quantitative only. Semiquantitative values should be used to estimate concentration for dilution purposes only. e. Analytical specificity: The cross-reactivity of parent drug, metabolites, and drugs commonly found in specimens was evaluated by adding known amounts of each substance to methadone metabolite-free urine. A compound producing negative results compared to the 100 ng/mL and 300 ng/mL cutoff calibrator was considered to have no cross-reactivity. The results of the study are presented below. Methadone, its metabolite, and structurally related compounds produced a negative result at the concentrations listed below. | Compound | Concentration, ng/mL | | --- | --- | | Methadone | 500,000 | | EMDP | 200,000 | | LAAM | 100,000 | | Nor-LAAM | 10,000 | Structurally unrelated compounds and/or concurrently used drugs produced a negative result at the concentrations listed below. | Compound | Conc. ng/mL | Compound | Conc. ng/mL | | --- | --- | --- | --- | | 6-AM | 500,000 | Imipramine | 1,000,000 | | Acetaminophen | 1,000,000 | Ketamine | 400,000 | | Acetylsalicylic acid | 1,000,000 | Levorphanol | 200,000 | | Amitriptyline | 100,000 | Levothyroxine | 500,000 | | Amoxicillin | 500,000 | Maprotiline | 1,000,000 | | Amphetamine | 1,000,000 | Meperidine | 1,000,000 | | Benzoylecgonine | 1,000,000 | d-Methamphetamine | 100,000 | | Caffeine | 100,000 | l-Methamphetamine | 100,000 | | Captopril | 500,000 | Metronidazole | 250,000 | | Carbamazepine | 500,000 | Morphine | 1,000,000 | | Chlordiazepoxide | 100,000 | Nalbuphine | 1,000,000 | | Chlorpromazine | 100,000 | Naloxone | 3,000,000 | | Cimetidine | 500,000 | Naltrexone | 3,000,000 | | Clomipramine | 100,000 | Norcodeine | 1,000,000 | | Cocaine | 200,000 | Normorphine | 1,000,000 | {6} The potential effect of endogenous and exogenous urine substances, pH, and specific gravity on the recovery of methadone metabolite using DRI Methadone Metabolite (100/300) Assay was assessed by spiking known amounts of potentially interfering substances into the negative and positive levels (± 25% of cutoffs of 100 and 300 ng/mL cutoff) for both cutoffs. The compounds were determined to not interfere at the concentrations shown below for the 100 and 300 ng/mL cutoffs: | Compound | Cmpd. Conc. | | --- | --- | | Acetaminophen | 100 ng/mL | | Acetone | 1 g/dL | | Ascorbic acid | 250 mg/dL | | Aspirin | 100 μg/mL | | Caffeine | 100 μg/mL | | Creatinine | 500 mg/dL | | Ethanol | 1 g/dL | | Galactose | 10 mg/dL | | γ-globulin | 500 mg/dL | | Glucose | 3 g/dL | | Hemoglobin | 150 mg/dL | | Human serum albumin | 500 mg/dL | | Ibuprofen | 100 μg/mL | | Oxalic Acid | 100 mg/dL | | pH range | 4-11 | | Specific gravity range | 1.004-1.035 | | Riboflavin | 7.5 mg/dL | | Sodium chloride | 900 mg/dL | | Urea | 1.25 g/dL | | Codeine | 1,000,000 | Nortriptyline | 500,000 | | --- | --- | --- | --- | | Desipramine | 1,000,000 | Oxazepam | 500,000 | | Dextromethorphan | 30,000 | Oxycodone | 500,000 | | Diazepam | 30,000 | Phencyclidine | 50,000 | | Dihydrocodeine | 1,000,000 | Phenobarbital | 1,000,000 | | Diphenhydramine | 500,000 | Phentermine | 1,000,000 | | Disopyramide | 1,000,000 | Promethazine | 100,000 | | Doxepine | 200,000 | Propoxyphene | 50,000 | | Doxylamine | 500,000 | Ranitidine | 500,000 | | Ephedrine | 2,000,000 | Salicyluric acid | 500,000 | | Fentanyl | 200,000 | Secobarbital | 1,000,000 | | Fluoxetine | 1,000,000 | Talwin | 500,000 | | Fluphenazine | 500,000 | 11-Nor-Δ⁹-THC-9- | 10,000 | | Heroin | 1,000,000 | COOH | | | Hydrocodone | 200,000 | Thebaine | 100,000 | | Hydromorphone | 200,000 | Thioridazine | 150,000 | | Ibuprofen | 1,000,000 | Tramadol | 500,000 | {7} f. Assay cut-off: Analytical performance of the device around the claimed cutoffs is described in precision section (1 a.) above 2. Comparison studies: a. Method comparison with predicate device: One hundred unaltered clinical specimens were tested using the DRI Methadone Metabolite (100/300) Assay in both the qualitative and semi-quantitative modes, and GC/MS. The results are presented as follows: Qualitative Stratified Results | DRI | Low Negative by GC/MS (less than -50%) | Near Cutoff Negative by GC/MS (between -50% and cutoff) | Near Cutoff Positive by GC/MS (between cutoff and +50%) | High Positive by GC/MS (greater than +50%) | Percent Agreement with GC/MS | | --- | --- | --- | --- | --- | --- | | 100 ng/mL Cutoff | | | | | | | Positive | 0 | 1 | 6 | 50 | 93% | | Negative | 30 | 9 | 4 | 0 | 98% | | 300 ng/mL Cutoff | | | | | | | Positive | 0 | 0 | 9 | 30 | 100% | | Negative | 50 | 10 | 1 | 0 | 98% | GC/MS Summary of Discordant Qualitative Results | Cutoff Value (ng/mL) | DRI Result | GC/MS (ng/mL) | Major Drug Present by GC/MS | | --- | --- | --- | --- | | 100 | POS | 96 | EDDP | | 100 | NEG | 130 | EDDP | | 100 | NEG | 131 | EDDP | | 100 | NEG | 125 | EDDP | | 100 | NEG | 128 | EDDP | | 300 | NEG | 310 | EDDP | {8} Semi-quantitative Stratified Results | DRI | Low Negative by GC/MS (less than -50%) | Near Cutoff Negative by GC/MS (between -50% and cutoff) | Near Cutoff Positive by GC/MS (between cutoff and +50%) | High Positive by GC/MS (greater than +50%) | Percent Agreement with GC/MS | | --- | --- | --- | --- | --- | --- | | 100 ng/mL Cutoff | | | | | | | Positive | 0 | 0 | 9 | 50 | 98% | | Negative | 30 | 10 | *1 | 0 | 100% | | 300 ng/mL Cutoff | | | | | | | Positive | 0 | 0 | 10 | 30 | 100% | | Negative | 50 | 10 | 0 | 0 | 100% | *GC/MS Summary of Discordant Semi-quantitative Results | Cutoff Value (ng/mL) | DRI Result | GC/MS (ng/mL) | Major Drug Present by GC/MS | | --- | --- | --- | --- | | 100 | neg | 130 | EDDP | b. Matrix comparison: Not applicable; this device is for use with urine only. 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 Part 809.10. O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision.