LZI METHAMPHETAMINE ENZYME MMUNOASSAY, LZI METHAMPHETAMINE CALIBRATORS, LZI METHAMPHETAMINE CONTROLS

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k113661 B. Purpose for Submission: New Device C. Measurand: Methamphetamine D. Type of Test: Qualitative and Semi-Quantitative Enzyme Immunoassay E. Applicant: Lin-Zhi International, Inc. F. Proprietary and Established Names: LZI Methamphetamine Immunoassay LZI Methamphetamine Drug of Abuse Calibrators LZI Methamphetamine Drug of Abuse Controls G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | LAF | Class II | 21 CFR § 862.3610 Methamphetamine test system | Toxicology (91) | | DLJ | Class II | 21 CFR § 862.3200 Clinical toxicology calibrator | Toxicology (91) | | LAS | Class I, Reserved | 21 CFR § 862.3280 Clinical toxicology control material | Toxicology (91) | {1} H. Intended Use: 1. Intended use(s): See indications for use below. 2. Indication(s) for use: The LZI Methamphetamine Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of d-methamphetamine in human urine, at the cutoff value of 500 ng/mL. The assay is designed for professional use with a number of automated chemistry analyzers. The semi-quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of specimen for confirmation by a confirmatory method such as GCMS or LCMS or (2) permitting laboratories to establish quality control procedures. The LZI Methamphetamine Drugs of Abuse (DAU) Calibrators are for use as calibrators in the qualitative and semi-quantitative calibration of the LZI Methamphetamine Enzyme Immunoassay at a cutoff value of 500 ng/mL. The LZI Methamphetamine Drugs of Abuse (DAU) Controls are for use as assayed quality control materials to monitor the precision of the LZI Methamphetamine Enzyme Immunoassay at a cutoff value of 500 ng/mL. The assay provides only a 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 spectrometry (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 only. 4. Special instrument requirements: Performance data was provided for Hitachi 717 analyzer. The assay can be used on a clinical chemistry analyzer capable of measuring absorbance at 340 nanometers. I. Device Description: The LZI Methamphetamine Enzyme Immunoassay is a kit comprised of two reagents, {2} separately packed, but sold together in one kit (R1 and R2). | Reagent | Description | | --- | --- | | R1 | Contains mouse monoclonal anti-methamphetamine antibody, glucose-6-phosphate (G6P), nicotinamide adenine dinucleotide (NAD), stabilizers, and sodium azide as a preservative. | | R2 | Contains d-methamphetamine labeled with glucose-6-phosphate dehydrogenase (G6PDH) in buffer with sodium azide as a preservative. | The LZI Methamphetamine Enzyme Immunoassay calibrators and controls designated for use at 500 ng/mL cutoff contain 0, 250, 375, 500, 625, 1,000, and 2,000 ng/mL of d-methamphetamine in human urine with less than 0.1% sodium azide as preservative. ## J. Substantial Equivalence Information: 1. Predicate device name(s): - LZI Amphetamines 500 Homogenous Enzyme Immunoassay - LZI Amphetamines 500 Drugs of Abuse Calibrators - LZI Amphetamines 500 Drugs of Abuse Controls 2. Predicate 510(k) number(s): - k102210 3. Comparison with predicate: Similarities and Differences – Test Device | Item | Candidate Device | Predicate Device | | --- | --- | --- | | | LZI Methamphetamine Enzyme Immunoassay | LZI Amphetamines 500 Homogenous Enzyme Immunoassay (k102210) | {3} | Intended Use | The Methamphetamine Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of d-methamphetamine in human urine, at the cutoff value of 500 ng/mL. The assay is designed for professional use with a number of automated chemistry analyzers.The semi-quantitative mode is for purposes of (1) enabling laboratories to determine an appropriate dilution of specimen for confirmation by a confirmatory method such as GCMS or LCMS or (2) permitting laboratories to establish quality control procedures.The Methamphetamine Drugs of Abuse (DAU) Calibrators are for use as calibrators in the qualitative and semi-quantitative calibration of the Methamphetamine Enzyme Immunoassay at a cutoff value of 500 ng/mL.The Methamphetamine Drugs of Abuse (DAU) Controls are for use as assayed quality control materials to monitor the precision of the Methamphetamine Enzyme Immunoassay at a cutoff value of 500 ng/mL.The assay provides only a 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 spectrometry (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. | Same | | --- | --- | --- | | Analytes | d-methamphetamine | d-amphetamine and d-methamphetamine | | Assay Type | Qualitative and Semi-Quantitative | Same | | Cutoff value | 500 ng/mL | Same | | Sample | Urine | Same | {4} | Methodology | Enzyme Immunoassay (EIA) | Enzyme Linked Immunoassay(ELISA) | | --- | --- | --- | | Analyzer | Clinical chemistry analyzer capable of measuring absorbance at 340 nanometers | Same | | Detection Wavelength | 340 nanometers | Same | | Calibrators | Five Levels (0, 250, 500, 1000, and 2,000) | Same | | Controls | Two Levels (375 and 625 ng/mL) | Same | # K. Standard/Guidance Document Referenced (if applicable): CLSI EP5-A, Evaluation of Precision Performance of Clinical Chemistry Devices # L. Test Principle: The assay is an Enzyme Immunoassay (EIA) 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, methamphetamine-labeled G6PDH conjugate is bound to antibody, and the enzyme activity is inhibited. On the other hand, when free drug is present in the sample, antibody would bind to free drug, the unbound methamphetamine-labeled G6PDH then exhibits its maximal enzyme activity. Active enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at $340~\mathrm{nm}$ . # M. Performance Characteristics (if/when applicable): # 1. Analytical performance: # a. Precision/Reproducibility: Precision studies were conducted on the Hitachi 717 analyzer using samples containing methamphetamine. Studies were performed according to CLSI-EP5. Samples were prepared by spiking a negative human urine pool with d-methamphetamine at the following concentrations: zero drug $(-100\%)$ , $-75\%$ , $-50\%$ , and $-25\%$ below the cutoff, cutoff, and $+25\%$ , $+50\%$ , $+75\%$ , and $+100\%$ above the cutoff. Samples were tested in 2 replicates per run, 2 runs per day for 22 days, total $n = 88$ . Samples concentrations were confirmed by GC/MS. Results of the studies are presented below: {5} | Semi-Quantitative | | Within Run | | Total Precision | | | --- | --- | --- | --- | --- | --- | | Sample Concentration (ng/mL) | % of Cutoff | Number of Determinations | Immunoassay Result | Number of Determinations | Immunoassay Result | | 0 | -100% | 22 | 22 Negative | 88 | 88 Negative | | 125 | -75% | 22 | 22 Negative | 88 | 88 Negative | | 250 | -50% | 22 | 22 Negative | 88 | 88 Negative | | 375 | -25% | 22 | 22 Negative | 88 | 88 Negative | | 500 | Cutoff | 22 | 22 Positive | 88 | 85 Positive 3 Negative | | 625 | +25% | 22 | 22 Positive | 88 | 88 Positive | | 750 | +50% | 22 | 22 Positive | 88 | 88 Positive | | 875 | +75% | 22 | 22 Positive | 88 | 88 Positive | | 1000 | +100% | 22 | 22 Positive | 88 | 88 Positive | | Qualitative | | Within Run | | Total Precision | | | --- | --- | --- | --- | --- | --- | | Sample Concentration (ng/mL) | % of Cutoff | Number of Determinations | Immunoassay Result | Number of Determinations | Immunoassay Result | | 0 | -100% | 22 | 22 Negative | 88 | 88 Negative | | 125 | -75% | 22 | 22 Negative | 88 | 88 Negative | | 250 | -50% | 22 | 22 Negative | 88 | 88 Negative | | 375 | -25% | 22 | 22 Negative | 88 | 88 Negative | | 500 | Cutoff | 22 | 16 Positive 6 Negative | 88 | 62 Positive 26 Negative | | 625 | +25% | 22 | 22 Positive | 88 | 88 Positive | | 750 | +50% | 22 | 22 Positive | 88 | 88 Positive | | 875 | +75% | 22 | 22 Positive | 88 | 88 Positive | | 1000 | +100% | 22 | 22 Positive | 88 | 88 Positive | b. Linearity/assay reportable range: Linearity across the range was confirmed by serially diluting a spike urine pool containing d-methamphetamine to obtain the levels listed in the table below. Each sample was assayed in 10 replicates on the Hitachi 717 analyzer in semi-quantitative mode. The results were averaged and compared to the expected result and the percent recovery was calculated. Results are presented below: {6} | Expected Value (ng/mL) | Observed Value (ng/mL) | % Recovery | | --- | --- | --- | | 0 | 0 | | | 25 | 22.5 | 86.0 | | 150 | 130.6 | 87.0 | | 300 | 291.6 | 97.2 | | 400 | 403.1 | 100.8 | | 500 | 497.1 | 99.4 | | 600 | 591.6 | 98.6 | | 750 | 726.2 | 96.8 | | 1000 | 966.6 | 96.7 | | 1400 | 1345.1 | 96.1 | | 2000 | 1971.4 | 98.6 | Linear regression analysis of the results yielded the following: $$ y = 0.9791x - 2.8289, R^2 = 0.9996 $$ c. Traceability, Stability, Expected values (controls, calibrators, or methods): Calibrators and controls were previously cleared (k102210) under a different trade name. The calibrators and controls of this current submission and the predicate only differ in trade name. See k102210 for calibrators and controls traceability and stability information. Labeling indicates to not use assay reagents, calibrators, or control materials beyond the indicated expiration dates on labeled vials. d. Detection limit: Performance at low drug concentrations in the semi-quantitative assay was characterized by determination of recovery (see section M1b above). e. Analytical specificity: Possible interference from endogenous compounds was evaluated using pooled urine samples spiked with d-methamphetamine at levels $\pm 25\%$ of the $500~\mathrm{ng/mL}$ cutoff. Interference was evaluated on the Hitachi 717 analyzer. No positive or negative interference was observed when the following endogenous compounds were tested at the physiological levels indicated in the table: {7} | Compound | Concentration (mg/dL) | -25% d-methamphetamine (375 ng/mL) | +25% d-methamphetamine (625 ng/mL) | | --- | --- | --- | --- | | Acetone | 1000 | Negative | Positive | | Ascorbic Acid | 500 | Negative | Positive | | Creatinine | 500 | Negative | Positive | | Ethanol | 1000 | Negative | Positive | | Galactose | 10 | Negative | Positive | | γ-Globulin | 500 | Negative | Positive | | Glucose | 1500 | Negative | Positive | | Hemoglobin | 100 | Negative | Positive | | Human Serum Albumin | 500 | Negative | Positive | | Oxalic Acid | 100 | Negative | Positive | | Riboflavin | 2.5 | Negative | Positive | | Sodium Chloride | 2000 | Negative | Positive | | Urea | 2000 | Negative | Positive | Possible interference from specific gravity was evaluated using urine samples containing d-methamphetamine at levels $\pm 25\%$ of the $500~\mathrm{ng/mL}$ cutoff with specific gravities ranging from 1.002 to 1.030. No positive or negative interference due to specific gravity was observed. To test for potential positive or negative interference from pH using urine samples containing d-methamphetamine at levels $\pm 25\%$ of the $500~\mathrm{ng/mL}$ cutoff were evaluated at pH values of 3, 4, 5, 6, 7, 8, 9, 10, and 11. No positive or negative interference due to pH was observed. Cross reactivity of various potential interfering structurally related drugs was tested by spiking a final concentration of up to $500,000~\mathrm{ng/mL}$ of each substance into drug-free urine. Final concentrations of drug compounds tested were equivalent to the $500~\mathrm{ng/mL}$ d-methamphetamine cutoff. Cross-reactivity was evaluated with the assay's calibrated dose-response curve. Results from these studies are summarized below: {8} | Structurally Related Compounds | Target Concentration (ng/mL) | Response equivalent to cutoff (ng/mL) | % Cross Reactivity | | --- | --- | --- | --- | | d-Amphetamine | 10,000 | 253.85 | 2.54% | | l-Amphetamine | 12,000 | 122.20 | 1.02% | | Atomoxetine | 500,000 | 129.60 | 0.03% | | Benzphetamine | 500,000 | 172.85 | 0.03% | | d-Ephedrine | 150,000 | 487.00 | 0.32% | | d,l-Ephedrine | 200,000 | 417.95 | 0.21% | | l-Ephedrine | 100,000 | 350.20 | 0.35% | | Fenfluramine | 4,000 | 433.30 | 10.83% | | 4-Fluoromethcathinone (Flephedrone; 4-MC) | 200,000 | 311.05 | 0.16% | | 3-Hydroxy-Tyramine | 500,000 | 250.55 | 0.04% | | Isoxsuprine | 500,000 | 147.85 | 0.03% | | Mephentermine | 25,000 | 107.50 | 0.43% | | l-Methamphetamine | 5,000 | 486.45 | 9.73% | | para-Methoxyamphetamine (PMA) | 400 | 12.75 | 3.19% | | para-Methoxymethylamphetamine (PMMA) | 500 | 374.70 | 74.94% | | Methylenedioxyamphetamine (MDA) | 1,400 | 28.60 | 2.04% | | Methylenedioxyethylamphetamine (MDEA) | 10,000 | 350.15 | 3.50% | | Methylenedioxymethylamphetamine (MDMA) | 1,000 | 389.10 | 38.91% | | 4-Methylmethcathinone (Mephedrone; 4-MMC; PMMC) | 100,000 | 393.40 | 0.39% | | Phendimetrazine | 150,000 | 300.35 | 0.20% | | Phenethylamine | 25,000 | 294.55 | 1.18% | | Phenmetrazine | 40,000 | 418.45 | 1.05% | | Phentermine | 20,000 | 40.20 | 0.20% | | phenylephrine | 300,000 | 467.60 | 0.16% | | d,l-Phenylpropanolamine | 150,000 | 92.30 | 0.06% | | d-Pseudoephedrine | 112,500 | 441.25 | 0.39% | | l-Pseudoehedrine | 200,000 | 178.25 | 0.09% | | Tranylcypromine | 50,000 | 286.70 | 0.57% | | Tyramine | 400,000 | 350.05 | 0.09% | Cross-reactivity of non-structurally related compounds was evaluated by testing compounds spiked into urine samples containing d-methamphetamine at levels $\pm 25\%$ of the $500~\mathrm{ng/mL}$ cutoff. Non-structurally related compounds were tested at the concentration levels indicated in the table below. Cross-reactivity was evaluated with the assay's calibrated dose-response curve. Results from these studies are summarized below: {9} | Non-Structurally Related Compounds | Target Concentration (ng/mL) | Response equivalent to cutoff (ng/mL) | % Cross Reactivity | | --- | --- | --- | --- | | Acetaminophen | 400,000 | 2.9 | 0.001 | | Acetylsalicylic acid | 500,000 | 7.4 | 0.001 | | Amoharbital | 250,000 | 72.3 | 0.029 | | Benzoylecgonine | 250,000 | 75.2 | 0.030 | | Bromopheniramine | 250,000 | 100.3 | 0.040 | | Burpropion | 100,000 | 90.6 | 0.091 | | Buspiron | 125,000 | 86.1 | 0.069 | | Caffeine | 500,000 | 9.1 | 0.002 | | Chlorpheniramine | 250,000 | 47.2 | 0.019 | | Chlorpromazine | 250,000 | 99.6 | 0.040 | | Codeine | 250,000 | 79.5 | 0.032 | | Dextromethorphan | 500,000 | 11.4 | 0.002 | | Doxepine | 200,000 | 11.5 | 0.006 | | Meperidine | 250,000 | 79.6 | 0.032 | | Methadone | 250,000 | 96.5 | 0.039 | | Methapyrilene | 100,000 | 89.8 | 0.090 | | Methaqualone | 250,000 | 81.7 | 0.033 | | Morphine | 500,000 | 8.5 | 0.002 | | Oxazepam | 250,000 | 85.5 | 0.034 | | Phencyclidine | 500,000 | 101.1 | 0.020 | | Phenobarbital | 250,000 | 74.5 | 0.030 | | Phenothiazine | 50,000 | 24.5 | 0.049 | | Procainamide | 30,000 | 105.3 | 0.351 | | Promethazine | 250,000 | 44.6 | 0.018 | | Propoxyphene | 250,000 | 76.8 | 0.031 | | Propranolol | 250,000 | 85.2 | 0.034 | | Ranitidine | 5,000 | 178.0 | 3.559 | | Scopolamine | 250,000 | 77.5 | 0.031 | | Secobarbital | 250,000 | 74.9 | 0.030 | | Sertraline | 125,000 | 106.9 | 0.085 | | Thioridazine | 250,000 | 99.8 | 0.040 | | Trazodone | 50,000 | 81.8 | 0.164 | | Trifluoperazine | 125,000 | 67.9 | 0.054 | | Trifluopromazine | 125,000 | 65.2 | 0.052 | | Valproic Acid | 500,000 | 14.8 | 0.003 | f. Assay cut-off: There is a 500 ng/mL cutoff concentration claimed for this assay. See section M1a above for performance data around the cutoff. 2. Comparison studies: a. Method comparison with predicate device: The sponsor conducted a method comparison study to evaluate the performance of the device for detection of d-methamphetamine by testing 95 unaltered samples (48 negative and 47 positive samples) on the Hitachi 717 analyzer. Results were compared to results obtained with GC/MS or LC/MS. Studies were conducted in semi-quantitative and qualitative modes. Equivalent results were obtained for both modes. Study results are summarized below: {10} 11 | Method Comparison Results (500 ng/mL Cutoff) | | | | | | | --- | --- | --- | --- | --- | --- | | Candidate Device Results | Negative | < 50% of the cutoff (1-250 ng/mL) | Near Negative Cutoff (250-500 ng/mL) | Near Positive Cutoff (500-750 ng/mL) | High Positive (>750 ng/mL) | | Positive (47 samples) | 0 | 0 | 0 | 9 | 37 | | Negative (48 samples) | 20 | 16 | 12 | 1 | 0 | | Discordant Results | | | --- | --- | | Candidate Device Result | GC/MS or LC/MS Result | | Negative | 654 ng/mL | 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. 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.