LZI OPIATE 2000 HOMOGENEOUS ENZYME IMMUNOASSAY, LZI OPIATE 2000 CALIBRATORS, LZI OPIATE 2000 CONTROLS

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k120761 B. Purpose for Submission: New Device C. Measurand: Opiates D. Type of Test: Qualitative and semi-quantitative enzyme immunoassay E. Applicant: Lin-Zhi International, Inc. F. Proprietary and Established Names: LZI Opiate 2000 Enzyme Immunoassay LZI Opiate 2000 Enzyme Calibrators LZI Opiate 2000 Enzyme Controls G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | DJG | Class II | 21CFR 862.3650 Opiate Test System | Toxicology (91) | | DLJ | Class II | 21CFR 862.3200 Clinical Toxicology Calibrator | Toxicology (91) | | LAS | Class I, Reserved | 21CFR 862.3280 Clinical Toxicology Control Material | Toxicology (91) | {1} H. Intended Use: 1. Intended use(s): Please see indications for use below. 2. Indication(s) for use: The LZI Opiate 2000 Immunoassay is intended for the qualitative and semi-quantitative determination of morphine in human urine at a cutoff value of 2000 ng/mL. The assay is designated for professional use with a number of automated clinical chemistry analyzers. The semi-quantitative mode in for purpose of (1) enabling laboratories to determine an appropriate dilution for the specimen for confirmation by a confirmatory method such as GC/MS and LC/MS or (2) permitting laboratories to establish quality control procedures. LZI Opiate 2000 Enzyme Calibrators are for use as calibrators in the qualitative and semi-quantitative calibrations of the LZI Opiate 2000 Immunoassay at a cut off value of 2000 ng/mL. LZI Opiate 2000 Enzyme Controls are for use as assayed quality control material to monitor the precision of the LZI Opiate 2000 Immunoassay at a cut off value of 2000 ng/mL. The assay provides only preliminary analytical result. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or liquid chromatography/mass spectroscopy (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 test result is positive. 3. Special conditions for use statement(s): For prescription use. The assay provides only preliminary analytical test results. A more specific alternative chemical method must be used in order to obtain a confirmed analytical result. Gas or liquid chromatography/mass spectroscopy (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 test result is positive. 4. Special instrument requirements: Testing was performed on the Hitachi 717 automatic analyzer. 2 {2} I. Device Description: The LZI Opiate 2000 assay is a homogeneous enzyme immunoassay with two ready-to-use liquid reagents, and R1 and R2. These reagents are bottled separately but sold together within a kit. R1 solution contains 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-dehidrogenase (G6PDH) labeled with morphine in buffer with sodium azide (0.09%) as a preservative. The LZI Opiate 2000 Immunoassay calibrators designated for use with the 2000ng/mL cutoff contains 5 levels, 0, 1000, 2000, 4000 and 6000 ng/mL of morphine in human urine with sodium azide (0.09%) as a preservative. These are sold as individual bottles. The LZI Opiate 2000 Immunoassay controls designated for use with the 2000ng/mL cutoff contains 2 levels, 1500 and 2500 ng/mL of morphine in human urine with sodium azide (0.09%) as a preservative. These are sold as individual bottles. J. Substantial Equivalence Information: 1. Predicate device name(s): LZI Opiate Enzyme Immunoassay Opiate Calibrators and Control 2. Predicate 510(k) number(s): k020368 and k020769 3. Comparison with predicate: | Item | LZI Opiate 2000 Immunoassay (Candidate Device) | LZI Opiate Enzyme Immunoassay (Predicate - k020368) | | --- | --- | --- | | Indication for Use | The LZI Opiate 2000 Immunoassay is intended for the qualitative and semi-quantitative determination of morphine in human urine at a cutoff value of 2000 ng/mL. The assay is designated for professional use with a number of automated clinical chemistry analyzers. | Same | | Test System | Hitachi 717 automatic analyzer | Same | {3} | Analyte | Morphine | Same | | --- | --- | --- | | Cutoff Concentration | 2000 ng/mL | 300 ng/mL | | Test Principle | Enzyme Immunoassay (EIA) | Same | | Matrix | Urine | Same | | Confirmation Method | LC/MS/MS | Same | | Storage | 2-8 °C until expiration date | Same | | Item | LZI Opiate 2000 Enzyme Calibrators (Candidate Device) | LZI Opiate Enzyme Calibrators (Predicate- k020769) | | --- | --- | --- | | Indication for Use | LZI Opiate 2000 Enzyme Calibrators are for use as calibrators in the qualitative and semi-quantitative calibrations of the LZI Opiate 2000 Immunoassay at a cut off value of 2000 ng/mL. | Same | | Levels | 0, 1000, 2000, 4000 and 6000 ng/mL | 0, 150, 300, 600 and 1000 ng/mL | | Item | LZI Opiate 2000 Enzyme Controls (Candidate Device) | LZI Opiate Enzyme Controls (Predicate- k020769) | | Indication for Use | LZI Opiate 2000 Enzyme Control are for use as assayed quality control material to monitor the precision of the LZI Opiate 2000 Immunoassay at a cut off value of 2000 ng/mL. | Same | | Levels | 1500 and 2500 ng/mL | 225 and 375 ng/mL | # K. Standard/Guidance Document Referenced (if applicable): CLSI Evaluation Protocols: EP5-A Evaluation of precision performance of clinical chemistry devices. # L. Test Principle: The LZI Opiate 2000 Immunoassay is based on competition between drug in the sample and the 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 the drug in the sample, morphine-labeled G6PDH conjugate is bound to the antibody, and the enzyme activity is inhibited. When free drug is present in the sample, the antibody binds to the free drug; the unbound morphine-labeled G6PDH then exhibits maximal enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at $340\mathrm{nm}$ . {4} M. Performance Characteristics (if/when applicable): 1. Analytical performance: a. Precision/Reproducibility: 1. Intra-assay precision around the cutoff Urine samples containing morphine at concentrations 0, 500, 1000, 1500, 2000, 2500, 3000, 3500 and 4000 ng/mL were tested using two replicates per run, two runs per day for 22 days on the Hitachi 717 automatic analyzer. One experienced technician performed the analyses. The data is presented below. Qualitative: | Sample concentration (ng/mL) | Within Run | | Between Run | | | --- | --- | --- | --- | --- | | | No. Observations | Immunoassay Results | No. Observations | Immunoassay Results | | 0 | 22 | 22 Negative | 88 | 88 Negatives | | 500 | 22 | 22 Negative | 88 | 88 Negatives | | 1000 | 22 | 22 Negative | 88 | 88 Negatives | | 1500 | 22 | 22 Negative | 88 | 88 Negatives | | 2000 (cutoff) | 22 | 9 Positives / 13 Negatives | 88 | 33 Positives / 55 Negatives | | 2500 | 22 | 22 Positives | 88 | 88Positives | | 3000 | 22 | 22 Positives | 88 | 88Positives | | 3500 | 22 | 22 Positives | 88 | 88Positives | | 4000 | 22 | 22 Positives | 88 | 88Positives | Semi-Quantitative: | Sample concentration (ng/mL) | Within Run | | Between Run | | | --- | --- | --- | --- | --- | | | No. Observations | Immunoassay Results | No. Observations | Immunoassay Results | | 0 | 22 | 22 Negative | 88 | 88 Negatives | | 500 | 22 | 22 Negative | 88 | 88 Negatives | | 1000 | 22 | 22 Negative | 88 | 88 Negatives | | 1500 | 22 | 22 Negative | 88 | 88 Negatives | | 2000 (cutoff) | 22 | 18 Positives / 4 Negatives | 88 | 59 Positives / 29 Negatives | | 2500 | 22 | 22 Positives | 88 | 88Positives | | 3000 | 22 | 22 Positives | 88 | 88Positives | | 3500 | 22 | 22 Positives | 88 | 88Positives | | 4000 | 22 | 22 Positives | 88 | 88Positives | {5} b. Linearity/assay reportable range: Linearity across the range was confirmed by using a morphine standard obtained from a commercial vendor and diluted several times using a pool of processed negative urine to create a stock solution with a concentration 6000 ng/mL of morphine. This stock was then diluted into twelve levels as presented in the table below. Each sample was tested in replicates of 10 on the Hitachi 717 analyzer. | % Dilution | Expected Concentration (ng/mL) | Observed Concentration (ng/mL) | % Recovery | | --- | --- | --- | --- | | 100 | 6000 | 6811.48 | 113.5 | | 90 | 5400 | 5837.71 | 108.1 | | 80 | 4800 | 4881.50 | 101.7 | | 70 | 4200 | 4239.37 | 100.9 | | 60 | 3600 | 3622.10 | 100.6 | | 50 | 3000 | 3007.99 | 100.3 | | 40 | 2400 | 2380.17 | 99.2 | | 30 | 1800 | 1842.90 | 102.4 | | 20 | 1200 | 1252.90 | 104.4 | | 10 | 600 | 596.78 | 99.5 | | 3.33 | 200 | 214.43 | 107.2 | | 0 | 0 | 80.72 | not applicable | c. Traceability, Stability, Expected values (controls, calibrators, or methods): Five levels of calibrators (0, 1000, 2000, 4000 and 6000 ng/mL) and two levels of control material (1500 and 2500 ng/mL) are available for use with the LZI Opiate 2000 Immunoassay. A commercially available morphine standard solution is used and traceable to NIST standard. This standard solution is made into a secondary (lower concentration) stock solution. The secondary stock solution is then spiked into the calibrators and controls to the desired concentration. The concentrations are confirmed by GC/MS. Stability Studies: Accelerated and long term studies for both controls and calibrators have been conducted. Protocols and acceptance criteria were described and found to be acceptable. The manufacturer claims the following expiration date for both controls and calibrators: When stored at 2-8 °C unopened or opened product is stable until expiration date which is 18 months. {6} 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: Cross-reactivity was established for the qualitative and semi-quantitative mode by spiking various concentrations of each substance into drug-free urine and then calibrated with the assay's calibrated dose-response curve. Results are expressed as a minimum concentration of metabolite or compound required to produce a response approximately equivalent to the cutoff concentration of the assay. The percent cross-reactivity of those compounds is presented below: Structurally related compounds | Compound | Target Concentration (ng/mL) | % Cross-reactivity | | --- | --- | --- | | 6-Monoacetyl Morphine | 2,000 | 111.74 | | Codeine | 1,900 | 113.92 | | Dihydrocodeine | 7,000 | 31.81 | | Heroin | 2,600 | 79.97 | | Hydrocodone | 15,000 | 15.64 | | Hydromorphone | 12,500 | 18.83 | | Levorphanol | 56,000 | 3.88 | | Morphine | 2,000 | 101.84 | | Morphine-3-Glucuronide (M3G) | 5,000 | 40.06 | | Morphine-6-Glucuronide (M6G) | 2,500 | 83.93 | | Norcodeine | 305,000 | 0.66 | | Oxycodone | 600,000 | 0.37 | | Oxymorphone | 1,020,000 | 0.20 | | Thebaine | 15,000 | 13.49 | | Codeine-6-B-Glucoronide | 2,000 | 107.95 | | Norhydrocodone | 1,000,000 | 0.15 | | JHydromorphone-3B-D-Glucuronide | 50,000 | 4.14 | {7} Structurally unrelated | Compound | Target Concentration (ng/mL) | -25% Morphine Cutoff | +25% Morphine Cutoff | | --- | --- | --- | --- | | Acetaminophen | 3,000,000 | Negative | Positive | | Acetylsalicylic Acid | 2,000,000 | Negative | Positive | | Albuterol | 70,000 | Negative | Positive | | Amitriptyline | 25,000 | Negative | Positive | | Amobarbital | 300,000 | Negative | Positive | | d-Amphetamine | 3,000,000 | Negative | Positive | | Benzoylecgonine | 3,000,000 | Negative | Positive | | Bupropion | 3,000,000 | Negative | Positive | | Caffeine | 1,500,000 | Negative | Positive | | Carbamazepine | 3,000,000 | Negative | Positive | | Chlorpomazine | 5,000 | Negative | Positive | | Clomipramine | 500,000 | Negative | Positive | | Desipramine | 1,000 | Negative | Positive | | Dextromethorphan | 400,000 | Negative | Positive | | Doxepine | 50,000 | Negative | Positive | | Ecgonine | 3,000,000 | Negative | Positive | | Ephedrine | 3,000,000 | Negative | Positive | | Fentanyl | 300 | Negative | Positive | | Fluoxetine | 60,000 | Negative | Positive | | Fluphenazine | 750,000 | Negative | Positive | | Ibuprofen | 3,000,000 | Negative | Positive | | Imipramine | 200,000 | Negative | Positive | | Lidocaine | 60,000 | Negative | Positive | | Maprotiline | 75,000 | Negative | Positive | | Meperidine | 30,000 | Negative | Positive | | Methadone | 400,000 | Negative | Positive | | Methapyrilene | 600,000 | Negative | Positive | | Methaqualone | 51,000 | Negative | Positive | | Metronidazole | 700,000 | Negative | Positive | | Nalbuphine | 30,000 | Negative | Positive | | Naloxone | 9,000 | Negative | Positive | | Naltrexone | 1,200,000 | Negative | Positive | | Nicotine | 10,000 | Negative | Positive | | Normorphine | 30,000 | Negative | Positive | | Nortriptyline | 360,000 | Negative | Positive | | Oxazepam | 3,000,000 | Negative | Positive | | Phencyclidine | 360,000 | Negative | Positive | | Phenobarbital | 120,000 | Negative | Positive | | Propoxyphene | 110,000 | Negative | Positive | | Ranitidine | 318,000 | Negative | Positive | | Secobarbital | 100,000 | Negative | Positive | {8} 9 | Compound | Target Concentration (ng/mL) | -25% Morphine Cutoff | +25% Morphine Cutoff | | --- | --- | --- | --- | | Talwin | 100,000 | Negative | Positive | | Thioridazine | 6,000 | Negative | Positive | | Tramadol | 330,000 | Negative | Positive | | Valproic Acid | 2,000,000 | Negative | Positive | ## Endogenous Compounds The following endogenous compounds were added into drug-free urine, urine sample spiked to -25% of morphine and spiked to +25% of Morphine at various concentrations. The substances listed in the table below were determined not to interfere at the concentration shown: | Interfering Substance | Concentration Tested ng/mL | -25% Morphine Cutoff | +25% Morphine Cutoff | | --- | --- | --- | --- | | Acetone | 1000 | Negative | Positive | | Ascorbic Acid | 1500 | Negative | Positive | | Creatinine | 500 | Negative | Positive | | Ethanol | 1000 | Negative | Positive | | Galactose | 10 | Negative | Positive | | γ-Globulin | 500 | Negative | Positive | | Glucose | 3000 | Negative | Positive | | Hemoglobin | 300 | Negative | Positive | | Human Serum Albumin | 500 | Negative | Positive | | Oxalic Acid | 100 | Negative | Positive | | Riboflavin | 0.3 | Negative | Positive | | Sodium Chloride | 6000 | Negative | Positive | | Urea | 6000 | Negative | Positive | There is the possibility that other substances and/or factors not listed above may interfere with the test and cause false results, e.g., technical or procedural errors. ## pH and Specific Gravity To test for possible positive and/or negative interference from pH urine samples having pH from 3, 4, 5, 6, 7, 8, 9, 10 and 11 were used. Each of these samples were spiked with commercially available standard to -25% of the cutoff and +25% of the cutoff and evaluated against the assay's calibration curve. No positive or negative interference due to pH was observed. To test for possible positive and/or negative interference from specific gravity urine {9} samples having specific gravity from 1.000, 1.002, 1.005, 1.007, 1.010, 1.012, 1.015, 1.017, 1.020 and 1.022 were used. The samples were spiked to $-25\%$ of the cutoff and $125\%$ of the cutoff. No positive or negative interference due to specific gravity was observed. # f. Assay cut-off: Analytical performance of the device around the claimed cutoff is described in precision section (1 a.) above # 2. Comparison studies: # a. Method comparison with predicate device: One hundred and fifty unaltered clinical urine samples were evaluated by the Lin-Zhi Opiate 2000 Enzyme Immunoassay on the Hitachi 717 analyzer and compared to LC/MS. Results from the study are presented below: Semi-Quantitative: | Candidate Device Results | Negative | < 50% of cutoff concentration by LC/MS analysis | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration) | High Positive (greater than 50% above the cutoff concentration) | | --- | --- | --- | --- | --- | --- | | Positive | 0 | 0 | 8 | 19 | 33 | | Negative | 20 | 43 | 27 | 0 | 0 | $\%$ Agreement among positives is $100.00\%$ $\%$ Agreement among negatives is $91.67\%$ Discordant | Cutoff Value (ng/mL) | LZI Opiate Assay (POS/NEG) | Drug/Metabolite LC/MS value (ng/mL) | | --- | --- | --- | | 2000 | Positive | 1160 | | 2000 | Positive | 1708 | | 2000 | Positive | 1715 | | 2000 | Positive | 1726 | | 2000 | Positive | 1745 | | 2000 | Positive | 1854 | | 2000 | Positive | 1853 | | 2000 | Positive | 1869 | {10} 11 Qualitative | Candidate Device Results 2000 ng/mL Cutoff | Negative | < 50% of cutoff concentration by LC/MS analysis | Near Cutoff Negative (Between 50% below the cutoff and the cutoff concentration) | Near Cutoff Positive (Between the cutoff and 50% above the cutoff concentration) | High Positive (greater than 50% above the cutoff concentration) | | --- | --- | --- | --- | --- | --- | | Positive | 0 | 0 | 4 | 19 | 33 | | Negative | 20 | 43 | 31 | 0 | 0 | % Agreement among positives is 100.00% % Agreement among negatives is 95.83% Discordant | Cutoff Value (ng/mL) | LZI Opiate Assay (POS/NEG) | Drug/Metabolite LC/MS value (ng/mL) | | --- | --- | --- | | 2000 | Positive | 1708 | | 2000 | Positive | 1726 | | 2000 | Positive | 1853 | | 2000 | Positive | 1869 | b. Matrix comparison: Not applicable. 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 {11} 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