OPIATE ENZYME IMMUNOASSAY

{0} 1 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k110298 B. Purpose for Submission: New Device C. Measurand: Opiates D. Type of Test: Competitive enzyme immunoassay; qualitative and semi-quantitative E. Applicant: Lin-Zhi International, Inc. F. Proprietary and Established Names: Opiate Enzyme Immunoassay G. Regulatory Information: 1. Regulation section: 21 CFR § 862.3650 Opiate Test System 2. Classification: Class II 3. Product code: DJG, Enzyme Immunoassay, Opiate 4. Panel: 91 (Toxicology) H. Intended Use: {1} 1. Intended use(s): See Indications for use, below. 2. Indication(s) for use: The Opiate Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of opiates in human urine, at a cutoff value of 300 ng/mL when calibrated against morphine. The assay is designed for prescription use with a number of automated clinical chemistry analyzers. The semi-quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GCMS or (2) permitting laboratories to establish quality control procedures. This assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or liquid chromatography (GC/MS or LC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. 3. Special conditions for use statement(s): The assay is for prescription use 4. Special instrument requirements: Clinical chemistry analyzers capable of maintaining a constant temperature, pipetting sample, mixing reagent, measuring enzymatic rates at 340 nm and timing the reaction accurately can be used to perform this homogeneous immunoassay. Performance studies in this submission were performed on the Hitachi 717. I. Device Description: The Opiate Enzyme Immunoassay is a homogeneous enzyme immunoassay ready-to-use liquid reagent. The assay is comprised of two ready-to-use liquid reagents, R1 (1 x 100 mL or 1 x 1000 mL) and R2 (1 x 37.5 or 1 x 375 mL). R1 solution contains a mouse monoclonal anti-morphine antibody, glucose-6-phosphate (G6P), nicotinamide adenine dinucleotide (NAD), stabilizers, and sodium azide (0.09%) as a preservative. R2 solution contains glucose-6-phosphate dehydrogenase (G6PDH) labeled with morphine in buffer, with sodium azide (0.09%) as a preservative. The kits will be provided in two configurations, with R1 x 100 mL; R2 x 37.5 mL and with R1 x 1000 mL; R2 x 375 mL. {2} Previously cleared (k020769) Calibrators and Controls are sold separately and contain negative human urine with sodium azide as preservative. J. Substantial Equivalence Information: 1. Predicate device name(s): Opiate Enzyme Immunoassay 2. Predicate 510(k) number(s): k020368 3. Comparison with predicate: | Similarities and Differences | | | | --- | --- | --- | | Item | Candidate device (Opiate Enzyme Immunoassay) | Predicate device (k020368) | | Intended Use | The Opiate Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of opiates in human urine, at a cut off value of 300 mg/mL. The assay is designed for professional use with a number of automated clinical chemistry analyzers. The semi-quantitative mode is for purposes of: 1) enabling laboratories to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as GCMS or 2) permitting laboratories to establish quality control procedures. This assay provides only a preliminary analytical test result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or liquid chromatography (GC/MS or LC/MS) is the preferred | Same | {3} | Similarities and Differences | | | | --- | --- | --- | | | confirmatory method. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary result is positive. | | | Sample Types | Urine | Same | | Test Principle | Competitive enzyme immunoassay | Same | | Cutoff | 300 ng/mL | Same | K. Standard/Guidance Document Referenced (if applicable): No Standards/Guidance Referenced L. Test Principle: The Opiate Enzyme Immunoassay is a competitive enzyme immunoassay based on competition between drug in the sample and drug labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. Enzyme activity decreases upon binding to the antibody, and the drug concentration in the sample is measured in terms of enzyme activity. In the absence of drug in the sample, morphine-labeled G6PDH conjugate is bound to antibody and enzyme activity is inhibited. When free drug is present in the sample antibody binds to the free drug, the unbound morphine-labeled G6PDH then exhibits its maximum enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH resulting in an absorbance change measured spectrophotometrically at 340 nm. M. Performance Characteristics (if/when applicable): 1. Analytical performance: Performance was evaluated on the Hitachi 717 a. Precision/Reproducibility: Precision was determined by spiking morphine standards into drug free urine pool at various concentrations surrounding the cutoff: -100%, -75%, -50%, -25%, cutoff, +100%, +75%, +50%, +25% +100%. Concentrations were confirmed by GC/MS. Testing for both the with-in run and between-run studies were performed by testing each sample in replicate, with two runs per day, for 22 days. The qualitative and semi-quantitative results are presented below: {4} Qualitative Precision Data: | 300 ng/mL Cutoff Result | | Within Run | | Total Precision | | | --- | --- | --- | --- | --- | --- | | Sample Concentration (ng/mL) | % of Cutoff | Number of Determinations | Immunoassay Results | Number of Determinations | Immunoassay Results | | 0 | -100% | 22 | 22 Negative | 88 | 88 Negative | | 75 | -75% | 22 | 22 Negative | 88 | 88 Negative | | 150 | -50% | 22 | 22 Negative | 88 | 88 Negative | | 225 | -25% | 22 | 22 Negative | 88 | 88 Negative | | 300 | Cut off | 22 | 16 Pos/6 Neg | 88 | 51 Pos/37 Neg | | 375 | +25% | 22 | 22 Positive | 88 | 88 Positive | | 450 | +50% | 22 | 22 Positive | 88 | 88 Positive | | 525 | +75% | 22 | 22 Positive | 88 | 88 Positive | | 600 | +100% | 22 | 22 Positive | 88 | 88 Positive | Semi-Quantitative Precision Data: | 300 ng/mL Cutoff Result | | Within Run | | Total Precision | | | --- | --- | --- | --- | --- | --- | | Sample Concentration (ng/mL) | % of Cutoff | Number of Determinations | Immunoassay Results | Number of Determinations | Immunoassay Results | | 0 | -100% | 22 | 22 Negative | 88 | 88 Negative | | 75 | -75% | 22 | 22 Negative | 88 | 88 Negative | | 150 | -50% | 22 | 22 Negative | 88 | 88 Negative | | 225 | -25% | 22 | 22 Negative | 88 | 88 Negative | | 300 | Cut off | 22 | 6 Pos/16 Neg | 88 | 27 Pos/61 Neg | | 375 | +25% | 22 | 22 Positive | 88 | 88 Positive | | 450 | +50% | 22 | 22 Positive | 88 | 88 Positive | | 525 | +75% | 22 | 22 Positive | 88 | 88 Positive | | 600 | +100% | 22 | 22 Positive | 88 | 88 Positive | b. Linearity/assay reportable range: Linearity across the range was confirmed by serially diluting a spiked urine pool containing 1000 ng/mL of morphine to obtain the concentrations listed in the table below. Each sample was assayed in replicates of 10 on the Hitachi 717 analyzer in the semi-quantitative mode. The results were averaged and compared to the expected result and the percent recovery was calculated. Results are presented in the table below: | Expected Value (ng/mL) | Mean Observed Value (ng/mL) | Recovery (%) | | --- | --- | --- | | 1000 | 1099.09 | 109.9 | | 900 | 957.84 | 106.4 | {5} 6 | 800 | 832.13 | 104.0 | | --- | --- | --- | | 700 | 710.24 | 101.5 | | 600 | 619.26 | 103.2 | | 500 | 523.29 | 104.7 | | 400 | 425.71 | 106.4 | | 300 | 303.31 | 101.1 | | 200 | 225.04 | 112.5 | | 100 | 116.31 | 116.3 | | 20 | 20.81 | 104.1 | | 0 | 0 | Not Applicable | c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability: Five levels of calibrators (0, 150, 300, 600, 1000 ng/mL) and two levels of control material (225 and 375 ng/mL) are available for use with the Opiate Enzyme Immunoassay. These calibrator and control solutions are human urine based and were previously cleared under k020769 Stability and Value assignment: Stability and value assignment of the calibrators and controls was evaluated in k020769. d. Detection limit: Performance at low drug concentrations in the semi-quantitative assay was characterized by determination of recovery (see section b above). e. Analytical specificity: The cross-reactivity of various potential interfering drugs was tested by spiking a final concentration of up to 3,000,000 ng/mL of each substance into drug-free urine. The structurally un-related compounds were spiked into samples containing morphine at concentrations ±25% of the cutoff. Compounds that interfered with the performance of the device were further tested at ±50% of the cutoff concentration. Samples evaluated in both semi-quantitative and qualitative mode. Results summarize the approximate quantity of each compound that is equivalent in assay reactivity to the 300 ng/mL morphine cutoff and are presented below: Structurally Related Compounds | Compound | Compound Concentration | Cross-reactivity (%) | | --- | --- | --- | {6} 7 | | (ng/mL) | | | --- | --- | --- | | 6-Monoacetyl Morphine | 400 | 85.85 | | Codeine | 200 | 169.00 | | Dihydrocodeine | 700 | 43.44 | | Heroin | 300 | 107.88 | | Hydrocodone | 2300 | 13.74 | | Hydromorphone | 1900 | 17.23 | | Levorphanol | 8000 | 4.01 | | Morphine | 300 | 101.15 | | Morphine-3-Glucuronide | 800 | 39.12 | | Morphine-6-Glucuronide | 300 | 109.03 | | Nalbuphine | 3,000,000 | 0.006 | | Naloxone | 2,700,000 | 0.046 | | Naltrexone | 800,000 | 0.011 | | Normorphine | 30,000 | 0.316 | | Norcodeine | 130,000 | 0.24 | | Oxycodone | 60,000 | 0.51 | | Oxymorphone | 140,000 | 0.23 | | Thebaine | 2,000 | 16.12 | | Codeine-6-β-Glucuronide | 250 | 124.38 | **Structurally Unrelated Compounds** | Compound | Concentration (ng/mL) | -25% Cutoff | | +25% Cutoff | | | --- | --- | --- | --- | --- | --- | | | | Semi-quant | Qualitative | Semi-quant | Qualitative | | Acetaminophen | 500,000 | NEG | NEG | POS | POS | | Acetylsalicylic Acid | 3,000,000 | NEG | NEG | POS | POS | | Albuterol | 3,000,000 | NEG | NEG | POS | POS | | Amitriptyline | 50,000 | NEG | NEG | POS | POS | | Amobarbital | 3,000,000 | NEG | NEG | POS | POS | | d-Amphetamine | 3,000,000 | NEG | NEG | POS | POS | | Benzoylecgonine | 3,000,000 | NEG | NEG | POS | POS | | Bupropion | 1,000,000 | NEG | NEG | POS | POS | | Caffeine | 3,000,000 | NEG | NEG | POS | POS | | Carbamazepine | 500,000 | NEG | NEG | POS | POS | | Chlorpromazine | 80,000 | NEG | NEG | POS | POS | | Clomipramine | 30,000 | NEG | NEG | POS | POS | {7} The following compounds interfered with the performance of the assay at $+/- 25\%$ of the cutoff concentration of morphine: Imipramine, Methapyrilene, Ranitidine, Tramadol. These compounds were further tested at $+/- 50\%$ of the cutoff and were shown to have no detectable interference with the assay. # Endogenous Compounds: The following endogenous compounds were spiked into urine spiked with morphine to $\pm 25\%$ of cutoff (225 or $375~\mathrm{ng / mL}$ ). The spiked solutions were evaluated on the Hitachi 717. The substances listed in the following table were determined not to interfere at the concentrations tested: | Compound | Concentration (ng/mL) | -25% Cutoff | | +25% Cutoff | | | --- | --- | --- | --- | --- | --- | | | | Qual. | Semi- | Qual. | Semi- | | Imipramine | 225 | 375 | 375 | 375 | 375 | | Dextramethorphan | 40,000 | NEG | NEG | POS | POS | | Doxepine | 175,000 | NEG | NEG | POS | POS | | Ecgonine | 3,000,000 | NEG | NEG | POS | POS | | Ephedrine | 1,400,000 | NEG | NEG | POS | POS | | Fentanyl | 300,000 | NEG | NEG | POS | POS | | Fluoxetine | 800,000 | NEG | NEG | POS | POS | | Fluphenazine | 3,000,000 | NEG | NEG | POS | POS | | Ibuprofen | 500,000 | NEG | NEG | POS | POS | | Imipramine | 20,000 | POS | POS | POS | POS | | Lidocaine | 3,000,000 | NEG | NEG | POS | POS | | Maprotiline | 600,000 | NEG | NEG | POS | POS | | Meperidine | 25,000 | NEG | NEG | POS | POS | | Methadone | 700,000 | NEG | NEG | POS | POS | | Methapyrilene | 300,000 | POS | POS | POS | POS | | Methaqualone | 3,000,000 | NEG | NEG | POS | POS | | Metronidazole | 700,000 | NEG | NEG | POS | POS | | Nicotine | 800,000 | NEG | NEG | POS | POS | | Nortriptyline | 110,000 | NEG | NEG | POS | POS | | Oxazepam | 3,000,000 | NEG | NEG | POS | POS | | Phencyclidine | 900,000 | NEG | NEG | POS | POS | | Phenobarbital | 3,000,000 | NEG | NEG | POS | POS | | Propoxyphene | 260,000 | NEG | NEG | POS | POS | | Ranitidine | 3,000,000 | POS | NEG | POS | POS | | Secobarbital | 3,000,000 | NEG | NEG | POS | POS | | Talwin | 100,000 | NEG | NEG | POS | POS | | Thioridazine | 70,000 | NEG | NEG | POS | POS | | Tramadol | 500,000 | POS | NEG | POS | POS | | Valproic Acid | 3,000,000 | NEG | NEG | POS | POS | {8} | | | | quant | | quant | | --- | --- | --- | --- | --- | --- | | Acetone | 1000 | NEG | NEG | POS | POS | | Ascorbic Acid | 1500 | NEG | NEG | POS | POS | | Creatinine | 500 | NEG | NEG | POS | POS | | Ethanol | 1000 | NEG | NEG | POS | POS | | Galactose | 10 | NEG | NEG | POS | POS | | β-Globulin | 500 | NEG | NEG | POS | POS | | Glucose | 3000 | NEG | NEG | POS | POS | | Hemoglobin | 300 | NEG | NEG | POS | POS | | Human Serum Albumin | 500 | NEG | NEG | POS | POS | | Oxalic Acid | 100 | NEG | NEG | POS | POS | | Riboflavin | 0.3 | NEG | NEG | POS | POS | | Sodium Chloride | 6000 | NEG | NEG | POS | POS | | Urea | 6000 | NEG | NEG | POS | POS | The package insert includes a complete list of all structurally related, structurally un-related and endogenous compounds tested. **Specific gravity:** Eight drug-free urine samples with specific gravity ranging from of 1.002 to 1.027 (1.002, 1.005, 1.008, 1.010, 1.012, 1.018, 1.02, 1.027 were spiked with morphine to achieve concentrations of $\pm 25\%$ of the cut-off (225 ng/mL or 375 mg/mL). The original and the spiked samples were tested with Opiate Enzyme Immunoassay on the Hitachi 717 in the qualitative and semi-quantitative modes. The results indicate that there is no positive or negative interference due to specific gravity. **pH:** The effect of pH was tested across the range of 3 to 11 (pH 3, 4, 5, 6, 7, 8, 9, 10, and 11). Drug free urine sample pools were pH adjusted then divided into two aliquots and then spiked with morphine to achieve $\pm 25\%$ of the cutoff. No positive or negative interference due to pH was observed. **f. Assay cut-off:** See Detection Limit Section, above. **2. Comparison studies:** **a. Method comparison with predicate device:** A total of 88 unaltered clinical samples (44 positive and 44 negative) were tested with the Opiate Enzyme Immunoassay on the Hitachi 717 analyzer. The opiate concentrations for each sample were confirmed by GC/MS or LC/MS. Results are shown in the tables below: {9} Qualitative: | Candidate Device Results | Negative | Low Negative (<50% of the cutoff by GC/MS) | Near Cutoff Negative (50% below the cutoff to the cutoff by GC/MS) | Near Cutoff Positive (50% above the cutoff to the cutoff by GC/MS) | High Positive (>50 above the cutoff by GC/MS) | Percent Agreement with GCMS | | --- | --- | --- | --- | --- | --- | --- | | Positive | 0 | 1 | 3 | 9 | 34 | 90.9% | | Negative | 3 | 25 | 12 | 1 | 0 | 97.7% | Summary of Discordant Results (Qualitative) | Opiate Assay (POS/NEG) | Drug/Metabolite GC/MS or LC/MS value based on cross reactivity profile | | --- | --- | | Positive | 40.6 ng/mL (Morphine) | | Positive | 244.62 ng/mL (Morphine) | | Positive | 252.7 ng/mL (Morphine) | | Positive | 241.9 ng/mL (Morphine) | | Negative | 320.7 ng/mL (Morphine) | Semi-Quantitative: | Candidate Device Results | Negative | Low Negative (<50% of the cutoff by GC/MS) | Near Cutoff Negative (50% below the cutoff to the cutoff by GC/MS) | Near Cutoff Positive (50% above the cutoff to the cutoff by GC/MS) | High Positive (>50 above the cutoff by GC/MS) | Percent Agreement with GCMS | | --- | --- | --- | --- | --- | --- | --- | | Positive | 0 | 0 | 3 | 9 | 34 | 93.18% | | Negative | 3 | 26 | 12 | 1 | 0 | 97.6% | Summary of Discordant Results (Semi-Quantitative) | Opiate Assay (POS/NEG) | Drug/Metabolite GC/MS or LC/MS value based on cross reactivity profile | | --- | --- | | Positive | 244.62 ng/mL (Morphine) | | Positive | 252.7 ng/mL (Morphine) | | Positive | 241.9 ng/mL (Morphine) | | Negative | 320.74 ng/mL (Morphine) | The tables above demonstrate that in this study qualitatively discrepant results were observed only for near-cutoff samples (+/- 50% of the cutoff concentration), {10} with the exception of 1 low negative sample that resulted in a positive result in the qualitative mode. A supplementary study, with an additional 42 unaltered patient samples was performed by the sponsor to support method comparison. Results from the additional study are presented below: Qualitative: | Candidate Device Results | Negative | Low Negative (<50% of the cutoff by GC/MS) | Near Cutoff Negative (50% below the cutoff to the cutoff by GC/MS) | Near Cutoff Positive (50% above the cutoff to the cutoff by GC/MS) | High Positive (>50 above the cutoff by GC/MS) | Percent Agreement with GCMS | | --- | --- | --- | --- | --- | --- | --- | | Positive | 0 | 0 | 0 | 7 | 6 | 100% | | Negative | 20 | 4 | 5 | 0 | 0 | 100% | Semi-Quantitative: | Candidate Device Results | Negative | Low Negative (<50% of the cutoff by GC/MS) | Near Cutoff Negative (50% below the cutoff to the cutoff by GC/MS) | Near Cutoff Positive (50% above the cutoff to the cutoff by GC/MS) | High Positive (>50 above the cutoff by GC/MS) | Percent Agreement with GCMS | | --- | --- | --- | --- | --- | --- | --- | | Positive | 0 | 0 | 0 | 7 | 6 | 100% | | Negative | 20 | 4 | 5 | 0 | 0 | 100% | No additional discrepant results were observed in the supplementary study. b. Matrix comparison: Not Applicable, urine is the only indicated matrix. 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. {11} 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. 12