CEDIA Heroin Metabolite (6-AM) Assay

{0} FDA U.S. FOOD &amp; DRUG ADMINISTRATION # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY ## I Background Information: A 510(k) Number K231007 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: New device. B Measurand: 6-Acetylmorphine (6-AM). C Type of Test: Qualitative and Semi-quantitative Homogeneous Enzyme Immunoassay ## III Intended Use/Indications for Use: A Intended Use(s): 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 Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993-0002 www.fda.gov {1} 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/ 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. ## B Indication(s) for Use: See Intended use above ## C Special Conditions for Use Statement(s): Rx - For Prescription Use Only ## D Special Instrument Requirements: Performance characteristics studies were conducted on the Horiba Yumizen C1200 clinical chemistry analyzer. ## IV Device/System Characteristics: ### A Device Description: CEDIA™ Heroin Metabolite (6-AM) assay is supplied as a two liquid (1 and 2) and two lyophilized reagents (1a and 2a) kit that is available in a kit configuration. 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 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 unique homogeneous enzyme immunoassay system. The assay is based on the bacterial enzyme β-galactosidase, which has been genetically engineered into two inactive fragments. These K231007 - Page 2 of 10 {2} 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 $\mathrm{CEDIA}^{\mathrm{TM}}$ Heroin Metabolite (6-AM), 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 $\beta$ -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): K192943 C Comparison with Predicate(s): | Device & Predicate Device(s): | K231007 | K192943 | | --- | --- | --- | | 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 | | Analyte | 6-Acetylmorphine (6-AM) | 6-Acetylmorphine (6-AM) | | General Device Characteristic Differences | | | | Instrument | Horiba Yumizen C1200 | Horiba Pentra C400 | K231007 - Page 3 of 10 {3} VI Standards/Guidance Documents Referenced: CLSI EP05-A3 – Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline – Third Edition. VII Performance Characteristics (if/when applicable): A Analytical Performance: All analytical performance studies were conducted on the Horiba Yumizen C1200 clinical chemistry analyzer. 1. Precision/Reproducibility: The precision study was performed based upon recommendations in CLSI Guideline EP05-A3, at one site with one analyzer and two lots of reagents. 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 6- Acetylmorphine analyte to final concentrations of -100%, -75%, -50%, -25%, below cutoff, cutoff, and +25%, +50%, +75% and +100%, above cutoff, and the concentrations were confirmed by LC-MS/MS or GC/MS. For qualitative and semi-quantitative mode results from a representative lot are summarized in the tables below. Precision Data in Qualitative Mode | % of Cutoff | Spiked Conc. (ng/mL) | # of Determinants | # Negative/# Positive | | --- | --- | --- | --- | | -100 | 0 | 80 | 80 / 0 | | -75 | 2.5 | 80 | 80 / 0 | | -50 | 5 | 80 | 80 / 0 | | -25 | 7.5 | 80 | 80 / 0 | | 100 | 10 | 80 | 36 / 44 | | +25 | 12.5 | 80 | 0 / 80 | | +50 | 15 | 80 | 0 / 80 | | +75 | 17.5 | 80 | 0 / 80 | | +100 | 20 | 80 | 0 / 80 | Precision Data in Semi-Quantitative Mode | % of Cutoff | Spiked Conc. (ng/mL) | # of Determinants | # Negative/# Positive | Within-run CV (%) | Total-run CV (%) | | --- | --- | --- | --- | --- | --- | | -100 | 0 | 80 | 80 / 0 | N/A | N/A | K231007 - Page 4 of 10 {4} K231007 - Page 5 of 10 | -75 | 2.5 | 80 | 80 / 0 | 9.5 | 14.5 | | --- | --- | --- | --- | --- | --- | | -50 | 5 | 80 | 80 / 0 | 3.9 | 6.2 | | -25 | 7.5 | 80 | 80 / 0 | 3.8 | 5.8 | | 100 | 10 | 80 | 31 / 49 | 3.6 | 5.4 | | +25 | 12.5 | 80 | 0 / 80 | 3.0 | 4.9 | | +50 | 15 | 80 | 0 / 80 | 3.4 | 4.9 | | +75 | 17.5 | 80 | 0 / 80 | 2.7 | 3.7 | | +100 | 20 | 80 | 0 / 80 | 1.9 | 3.3 | ## 2. 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-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: $$ y = 0.9008x + 0.3445; R^2 = 0.997 $$ The recovery of the samples from the linear range of the assay (2 ng/mL to 20 ng/mL) prepared by dilution ranged from 90.5% to 111.0%. ## 3. Analytical Specificity/Interference: A. Cross-reactivity (specificity) 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 was calculated as follows: % Cross-reactivity = (Cutoff concentration / Lowest concentration of cross reactant that gives a positive result) x 100 Cross-Reactivity of 6-Acetylmorphine and Heroin: | 6-Acetylmorphine and Heroin | Tested Concentration (ng/mL) | Assay Result | Cross-Reactivity (%) | | --- | --- | --- | --- | | 6-Acetylmorphine | 10 | Pos | 100% | | Heroin | 300 | Pos | 3% | Cross-Reactivity of Opiates and Structurally Related Compounds: {5} K231007 - Page 6 of 10 | Structurally related compounds and other opiates | Tested Concentration (ng/mL) | Assay Result | Cross-Reactivity (%) | | --- | --- | --- | --- | | 6-Acetylcodeine | 50,000 | Pos | 0.02% | | Buprenorphine | 100,000 | Neg | No Cross-Reactivity | | Buprenorphine-3β-D-glucuronide | 100,000 | Neg | No Cross-Reactivity | | Codeine | 100,000 | Neg | No Cross-Reactivity | | Dextromethorphan | 90,000 | Pos | 0.01% | | Dihydrocodeine | 100,000 | Neg | No Cross-Reactivity | | EDDP | 100,000 | Neg | No Cross-Reactivity | | EMDP | 100,000 | Neg | No Cross-Reactivity | | Ethylmorphine | 100,000 | Neg | No Cross-Reactivity | | Fentanyl | 100,000 | Neg | No Cross-Reactivity | | Hydrocodone | 100,000 | Neg | No Cross-Reactivity | | Hydromorphone | 20,000 | Pos | 0.04% | | Hydromorphone-3β-D-glucuronide | 100,000 | Neg | No Cross-Reactivity | | Levorphanol | 15,000 | Pos | 0.05% | | Methadone | 100,000 | Neg | No Cross-Reactivity | | Meperidine | 100,000 | Neg | No Cross-Reactivity | | Mitragynine | 100,000 | Neg | No Cross-Reactivity | | 7-Hydroxymitragynine | 100,000 | Neg | No Cross-Reactivity | | Morphine | 13,500 | Pos | 0.07% | | Morphine-3β-D-Glucuronide | 100,000 | Neg | No Cross-Reactivity | | Morphine-6β-D-Glucuronide | 100,000 | Neg | No Cross-Reactivity | | Nalorphine | 10,500 | Pos | 0.10% | | Naloxone | 100,000 | Neg | No Cross-Reactivity | | Naltrexone | 100,000 | Neg | No Cross-Reactivity | | Norbuprenorphine | 100,000 | Neg | No Cross-Reactivity | | Norbuprenorphine Glucuronide | 100,000 | Neg | No Cross-Reactivity | | Norcodeine | 100,000 | Neg | No Cross-Reactivity | | Norhydrocodone | 100,000 | Neg | No Cross-Reactivity | | Normorphine | 50,000 | Pos | 0.02% | | Norpropoxyphene | 100,000 | Neg | No Cross-Reactivity | | Noroxycodone | 100,000 | Neg | No Cross-Reactivity | | Noroxymorphone | 100,000 | Neg | No Cross-Reactivity | | Oxymorphone-3β-D-glucuronide | 100,000 | Neg | No Cross-Reactivity | | Oxycodone | 100,000 | Neg | No Cross-Reactivity | {6} B. Interference: The interference study was performed using one lot of reagents, calibrators and controls. Structurally unrelated compounds were evaluated by adding each substance to 6-Acetylmorphine spiked at low (7.5 ng/mL) and high (12.5 ng/mL) controls at the concentrations indicated. As shown in the tables below, all the pharmacologic compounds evaluated exhibited no cross-reactivity at the concentrations tested. - Samples were tested in replications of five (n=5) in both qualitative and semiquantitative modes. | Interferents | Spiked Concentration (ng/mL) | Spiked 6-Acetylmorphine Level | | | --- | --- | --- | --- | | | | Low Control | High Control | | 10,11 Dihydrocarbamazepine | 85,000 | Neg | Pos | | 11-nor-delta9-THC-COOH | 10,000 | Neg | Pos | | Acetaminophen | 500,000 | Neg | Pos | | Acetylsalicylic Acid | 500,000 | Neg | Pos | | Amitriptyline | 50,000 | Neg | Pos | | Amoxicillin | 500,000 | Neg | Pos | | Amphetamine | 100,000 | Neg | Pos | | Amisulpride | 100,000 | Neg | Pos | | Benztropine Mesylate | 50,000 | Neg | Pos | | Benzoylecgonine | 100,000 | Neg | Pos | | Brompheniramine | 75,000 | Neg | Pos | | Caffeine | 500,000 | Neg | Pos | | Captopril | 500,000 | Neg | Pos | | Chlordiazepoxide | 100,000 | Neg | Pos | | Chlorpromazine | 10,000 | Neg | Pos | | Clomipramine | 150,000 | Neg | Pos | | Chloroquine | 500,000 | Neg | Pos | | Cimetidine | 500,000 | Neg | Pos | | Desipramine | 50,000 | Neg | Pos | | Diazepam | 100,000 | Neg | Pos | | Digoxin | 100,000 | Neg | Pos | | Diphenhydramine | 20,000 | Neg | Pos | | Doxepin HCl | 10,000 | Neg | Pos | | Enalapril | 500,000 | Neg | Pos | | Fluoxetine | 500,000 | Neg | Pos | | Fluophenazine | 500,000 | Neg | Pos | | Haloperidol | 50,000 | Neg | Pos | K231007 - Page 7 of 10 {7} Interference with Endogenous Substances | Compound | Spiked Concentration (mg/dL) | Spiked 6-Acetylmorphine Level | | | --- | --- | --- | --- | | | | Low Control | High Control | | Acetone | 1000 | Neg | Pos | | Ascorbic acid | 1500 | Neg | Pos | | Creatinine | 500 | Neg | Pos | | Ethanol | 1000 | Neg | Pos | | Galactose | 10 | Neg | Pos | | Y-globulin | 500 | Neg | Pos | | Glucose | 1000 | Neg | Pos | | Hemoglobin | 300 | Neg | Pos | | Human serum albumin | 500 | Neg | Pos | | Oxalic acid | 100 | Neg | Pos | | Riboflavin | 7.5 | Neg | Pos | | Sodium Chloride | 6000 | Neg | Pos | | Urea | 2000 | Neg | Pos | | Hydroxychlroquine | 100,000 | Neg | Pos | | --- | --- | --- | --- | | Hydroxyzine | 125,000 | Neg | Pos | | Ibuprofen | 500,000 | Neg | Pos | | Imipramine | 20,000 | Neg | Pos | | Levothyroxine | 50,000 | Neg | Pos | | Methamphetamine | 100,000 | Neg | Pos | | Maprotiline | 500,000 | Neg | Pos | | Nalbuphine | 100,000 | Neg | Pos | | Naproxen | 500,000 | Neg | Pos | | Nortriptyline | 175,000 | Neg | Pos | | Nifedipine | 500,000 | Neg | Pos | | Nordiazepam | 60,000 | Neg | Pos | | Oxazepam | 100,000 | Neg | Pos | | Perphenazine | 150,000 | Neg | Pos | | Phencyclidine | 7,500 | Neg | Pos | | Phenobarbital | 100,000 | Neg | Pos | | Procyclidine | 150,000 | Neg | Pos | | Propoxyphene | 25,000 | Neg | Pos | | Protriptyline | 10,000 | Neg | Pos | | Ranitidine | 500,000 | Neg | Pos | | Salicyluric Acid | 500,000 | Neg | Pos | | Secobarbital | 100,000 | Neg | Pos | | Sulpiride | 500,000 | Neg | Pos | | Thioridazine | 250,000 | Neg | Pos | | Triprolidine | 125,000 | Neg | Pos | | Verapamil | 500,000 | Neg | Pos | K231007 - Page 8 of 10 {8} Effect of pH: Drug free urine spiked with 7.5 or 12.5 ng/mL of 6-Acetylmorphine was adjusted to the indicated pH (3, 4, 5, 6, 7, 8, 9, 10, and 11). Samples were tested in replicates of five (n=5) in both qualitative and semiquantitative modes. The results demonstrated that the tested range of pH did not affect the performance of the candidate test. ## Effect of Specific Gravity: Urine that spanned a specific gravity range of 1.000–1.030 were spiked with 7.5 or 12.5 ng/mL of 6-Acetylmorphine (6-AM). Samples were tested in replicates of five (n=5) in both qualitative and semiquantitative modes. The results demonstrated that the specific gravity range evaluated did not affect the performance of the candidate test. 4. Assay Reportable Range: Not applicable. 5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods): The device is traceable to a commercially available standard that was verified by LC/MS-MS or GC/MS. 6. Detection Limit: Not applicable. 7. Assay Cut-Off: See section VII A. 1 ## B Comparison Studies: Method Comparison with Predicate Device: A Method comparison study was performed using one hundred and twenty-three unaltered clinical samples analyzed using the candidate test in one replicate, in both qualitative and semi-quantitative modes. The results were compared to LC-MS/MS (Liquid chromatography-tandem mass spectroscopy). The results obtained in the qualitative and semi-quantitative modes are summarized below. Qualitative Mode: | CEDIA Heroin Metabolite (6-AM) Assay | < 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 – 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 | 6 | 53 | | Negative | 57 | 7 | 0 | 0 | K231007 - Page 9 of 10 {9} Semi-Quantitative Mode: | CEDIA Heroin Metabolite (6-AM) Assay | < 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 – 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 | 1 | 5 | 53 | | Negative | 57 | 6 | 1 | 0 | 1. 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. K231007 - Page 10 of 10