LZI Methadone Metabolite Enzyme Immunoassay, LZI Methadone Metabolite (100 and 300) Calibrators

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k170416 B. Purpose for Submission: New Device C. Measurand: EDDP (2-ethylidene-1,5-dimethyl-3,3- diphenylpyrrolidine) D. Type of Test: Qualitative and Semi-Quantitative Assay E. Applicant: Lin-Zhi International, Inc. F. Proprietary and Established Names: LZI Methadone Metabolite (EDDP) Enzyme Immunoassay LZI Methadone Metabolite (EDDP) (100 and 300) Calibrators G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | DJR | Class II | 862.3620 | 91-Toxicology | | DLJ | Class II | 862.3200 | 91-Toxicology | H. Intended Use: 1. Intended use(s): See indication for use below. {1} 2. Indication(s) for use: The LZI Methadone Metabolite (EDDP) Enzyme Immunoassay is an in vitro diagnostic test intended for the qualitative and semi-quantitative determination of Methadone Metabolite in human urine. The cutoff for both the qualitative and semi-quantitative modes of the assay are 100 ng/mL and 300 ng/mL for Methadone Metabolite. The assay is designed for prescription use on 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 Gas or liquid chromatography/mass spectrometry (GC/MS or LC/MS) or (2) permitting laboratories to establish quality control procedures. The assay provides only a preliminary analytical result. A more specific alternative analytical chemistry 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. The LZI Methadone Metabolite (EDDP) (100 and 300) Calibrators are for use as calibrators in the qualitative and semi-quantitative calibration of the LZI Methadone Metabolite (EDDP) Enzyme Immunoassay at the cutoff values of 100 ng/mL and 300 ng/mL. 3. Special conditions for use statement(s): For prescription use only 4. Special instrument requirements: Performance data was obtained using the Beckman Coulter AU480 Analyzer. I. Device Description: The LZI Methadone Metabolite (EDDP) Enzyme Immunoassay is comprised of two reagents, R1 and R2, which are provided as ready to use liquids. The R1 solution contains mouse monoclonal anti-methadone metabolite antibody, glucose-6-phosphate (G6P), nicotinamide adenine dinucleotide (NAD), stabilizers, and sodium azide (0.09 %) as a preservative. The R2 solution contains glucose-6-phosphate dehydrogenase (G6PDH) labeled with methadone metabolite in buffer with sodium azide (0.09 %) as a preservative. The LZI Methadone Metabolite (EDDP) 100 Calibrators contain 0, 50, 100, 250, and 500 ng/mL of methadone metabolite (EDDP) in human urine with sodium azide as a preservative. These five calibrators are sold as liquid, ready to use individual bottles. 2 {2} The LZI Methadone Metabolite (EDDP) 300 Calibrators contain 0, 150, 300, 650, and 1000 ng/mL of methadone metabolite (EDDP) in human urine with sodium azide as a preservative. These calibrators are sold as liquid, ready to use individual bottles. ## J. Substantial Equivalence Information: 1. Predicate device name(s): Lin-Zhi International, Inc Methadone Metabolite Enzyme Immunoassay 2. Predicate 510(k) number(s): k031797 3. Comparison with predicate: | Device Similarities | | | | --- | --- | --- | | Item | LZI Methadone Metabolite (EDDP) Enzyme Immunoassay (k170416) | Predicate Device The Lin-Zhi International, Inc. (LZI) Methadone Metabolite (EDDP) Enzyme Immunoassay (k031797) | | Intended use | The LZI Methadone Metabolite (EDDP) Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of methadone metabolite (EDDP) in human urine at cutoff values of 100 or 300 ng/mL. The assay is designed for professional use with a number of automated clinical chemistry analyzers. This assay provides a rapid screening procedure for determining the presence of methadone metabolite in urine. 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 | The Lin-Zhi International, Inc. (LZI) Methadone Metabolite (EDDP) Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of methadone metabolite in human urine at a cutoff value of 300 ng/mL. The assay is designed for professional use with a number of automated clinical chemistry analyzers. This assay provides a rapid screening procedure for determining the presence of methadone metabolite in urine. 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 | {3} | Device Similarities | | | | --- | --- | --- | | Item | LZI Methadone Metabolite (EDDP) Enzyme Immunoassay (k170416) | Predicate Device The Lin-Zhi International, Inc. (LZI) Methadone Metabolite (EDDP) Enzyme Immunoassay (k031797) | | | 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. | 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. | | Analyte | Methadone Metabolite (EDDP) | Same | | Matrix | Urine | Same | | Storage | 2-8°C until expiration date | Same | | Device Differences | | | | --- | --- | --- | | Item | LZI Methadone Metabolite (EDDP) Enzyme Immunoassay (k170416) | Predicate Device The Lin-Zhi International, Inc. (LZI) Methadone Metabolite (EDDP) Enzyme Immunoassay (k031797) | | Cutoff | 100 or 300 ng/ml | 300 ng/mL | {4} | Calibrators Similarities and Differences | | | | --- | --- | --- | | Item | LZI Methadone Metabolite (EDDP) Calibrators (k170416) | Predicate Device The Lin-Zhi International, Inc. (LZI) Single Analyte (EDDP) Drug of Abuse Calibrators (k031797) | | Intended use | The LZI Methadone Metabolite (EDDP) (100 and 300) Calibrators are for use as calibrators in the qualitative and semiquantitative calibration of the LZI Methadone Metabolite (EDDP) Enzyme Immunoassay at the cutoff values of 100 ng/mL and 300 ng/mL | The EDDP (methadone metabolite) Drugs of Abuse Calibrators are intended for in vitro diagnostic use for the calibration of the methadone metabolite enzyme immunoassays to detect methadone metabolite (EDDP) in human urine. | K. Standard/Guidance Document Referenced (if applicable): EP5-A, Evaluation of Precision Performance of Clinical Chemistry Devices, Vol. 19, No.2, February 1999. L. Test Principle: The LZI Methadone Metabolite assay is a homogeneous enzyme immunoassay based on competition between EDDP in the sample and EDDP labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for a fixed amount of antibody in the reagent. In the absence of EDDP in the sample, EDDP-labeled G6PDH conjugate is bound to antibody, and the G6PDH enzyme activity is inhibited. When free EDDP is present in the sample, it competes with EDDP-labeled G6PDH for binding to the antibody allowing for an amount of active G6PDH enzyme that is proportional to EDDP concentration. Active G6PDH enzyme converts nicotinamide adenine dinucleotide (NAD) to NADH, resulting in an absorbance change that can be measured spectrophotometrically at 340 nm. M. Performance Characteristics (if/when applicable): 1. Analytical performance: a. Precision/Reproducibility: Precision studies were carried out in qualitative and semi-quantitative mode. All methadone metabolite samples were tested in replicates of 2, two runs per day for 22 days on one Beckman Coulter AU480 automatic clinical analyzer for a total of 88 sample results. The samples were prepared by spiking drug into negative urine with the concentrations provided in the below tables. Each drug concentration was {5} confirmed by LC/MS. The results are summarized in the tables below, and were identical for each mode. Results are provided below. Qualitative and Semi-Quantitative results at 100 ng/mL Cutoff: | Methadone Metabolite (EDDP) concentration (ng/mL) | LC/MS Value (ng/ml) | % of cutoff | Within Run (N=22) | Total Precision (N=88) | | --- | --- | --- | --- | --- | | | | | Immunoassay Result | Immunoassay Result | | 0 | 0 | -100% | 22 Negative | 88 Negative | | 25 | 39 | -75% | 22 Negative | 88 Negative | | 50 | 57.2 | -50 % | 22 Negative | 88 Negative | | 75 | 76.6 | -25 % | 22 Negative | 88 Negative | | 100 | 103.5 | 0% | 11 Neg/11 Pos | 40 Pos/48 Neg | | 125 | 132 | +25% | 22 Positive | 88 Positive | | 150 | 165 | +50% | 22 Positive | 88 Positive | | 175 | 179 | +75% | 22 Positive | 88 Positive | | 200 | 192 | +100% | 22 Positive | 88 Positive | Qualitative and Semi-Quantitative results at 300 ng/mL Cutoff: | Methadone Metabolite (EDDP) concentration (ng/mL) | LC/MS Value (ng/ml) | % of cutoff | Within Run (N=22) | Total Precision | | --- | --- | --- | --- | --- | | | | | Immunoassay Result | Immunoassay Result | | 0 | 0 | -100% | 22 Negative | 88 Negative | | 75 | 76.6 | -75% | 22 Negative | 88 Negative | | 150 | 165 | -50 % | 22 Negative | 88 Negative | | 225 | 233 | -25 % | 22 Negative | 88 Negative | | 300 | 312 | 0% | 6 Neg/ 16 Pos | 36 Neg/ 52 Pos | | 375 | 391 | +25% | 22 Positive | 88 Positive | | 450 | 466 | +50% | 22 Positive | 88 Positive | | 525 | 541 | +75% | 22 Positive | 88 Positive | | 600 | 657 | +100 | 22 Positive | 88 Positive | b. Linearity/assay reportable range: A linearity study in the semi-quantitative mode was conducted by spiking a drug free urine pool to create a high concentration EDDP, and preparing serial dilutions with negative urine to achieve the EDDP concentrations provided in the tables below. Each level was tested in triplicate on the Beckman Coulter AU480 automated clinical chemistry analyzer. The concentrations were confirmed by LC/MS. The results of the linearity/recovery study are summarized below: {6} | Linearity: 100 ng/mL Cutoff | | | | --- | --- | --- | | Expected Concentration (ng/mL) | Observed Concentration (ng/mL) | Recovery (%) | | 0 | -3.1 | NA | | 10 | 5.2 | 51.7 | | 50 | 49.0 | 97.7 | | 100 | 98.8 | 98.8 | | 150 | 153.7 | 102.5 | | 200 | 203.0 | 101.5 | | 250 | 245.8 | 98.3 | | 300 | 312.7 | 104.2 | | 350 | 369.1 | 105.5 | | 400 | 412.9 | 103.2 | | 450 | 443.0 | 98.4 | | 500 | 478.2 | 95.6 | | Linearity: 300 ng/mL Cutoff | | | | --- | --- | --- | | Expected Concentration (ng/mL) | ObservedConcentration (ng/mL) | Recovery (%) | | 0 | -6.4 | NA | | 20 | 5.8 | 29.0 | | 100 | 83.1 | 83.1 | | 200 | 196.2 | 98.1 | | 300 | 295.9 | 96.6 | | 400 | 422.5 | 105.6 | | 500 | 519.0 | 103.8 | | 600 | 595.3 | 99.2 | | 700 | 709.4 | 101.3 | | 800 | 807.1 | 100.9 | | 900 | 878.4 | 97.6 | | 1000 | 940.9 | 94.1 | c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability and value assignment Calibrators are prepared from a standard solution of EDDP purchased from a commercial vendor. This standard solution is diluted with drug-free synthetic urine matrix. The concentrations of the prepared solutions are confirmed by LC/MS. {7} 8 # Stability Accelerated and real-time stability studies were conducted and the stability protocols and acceptance criteria were reviewed and found to be acceptable. The study results support the stability claim of eighteen (18) months when the LZI Methadone Metabolites EDDP Calibrators are stored at 2 to 8 °C. ## d. Detection limit: Not applicable. ## e. Analytical specificity: Compounds sharing structural similarity with EDDP were tested for cross-reactivity with the candidate device. The structurally related compounds that exhibited cross-reactivity with the candidate device were titrated to determine the lowest concentration that generated a positive result. The concentration (ng/mL) of cross-reactant that gives a response equivalent to the cutoff, and the calculated percent cross-reactivity are presented in the table below. Methadone Metabolite and Structurally Related Compounds for 100 ng/mL Cutoff: | Drug | Concentration (ng/mL) | Cross Reactivity (%) | | --- | --- | --- | | EDDP | 100 | 100 % | | EMDP | 100,000 | <0.1 % | | Methadone | 300,000 | <0.1 % | | LAAM HCl | 500,000 | <0.1 % | | (±)-α-Methadol | 300,000 | <0.1 % | | (-)-Isomethadone HCl | 60,000 | <0.1 % | | (-)-α-Noracetylmethadol (Nor-LAAM) HCl | 300,000 | <0.1 % | Methadone Metabolite and Structurally Related Compounds for 300 ng/mL Cutoff: | Drug | Concentration (ng/mL) | Cross Reactivity (%) | | --- | --- | --- | | EDDP | 300 | 100 % | | EMDP | 100,000 | <0.1 % | | Methadone | 500,000 | <0.1 % | | LAAM HCl | 500,000 | <0.1 % | | (±)-α-Methadol | 300,000 | <0.1 % | | (-)-Isomethadone HCl | 200,000 | <0.1 % | | (-)-α-Noracetylmethadol (Nor-LAAM) HCl | 300,000 | <0.1 % | {8} Potential interfering substances found in human urine of physiological or pathological conditions were added to drug-free urine. Samples were tested with Methadone Metabolite (EDDP) concentrations of 0 ng/mL, -25% of cut off and +25% of cut off. No interference was observed at the concentrations tested. Results are summarized below. Structurally Unrelated Pharmacological Compounds for 100 ng/mL Cutoff: | Interfering Substances | Maximun concentration tested without interference (ng/mL) | | --- | --- | | Acetaminophen | 100,000 | | 6-Acetylmorphine | 10,000 | | Acetylsalicylic Acid | 100,000 | | Alimemazine | 1,000 | | Amitriptyline | 100,000 | | Amlodipine | 100,000 | | Amoxicillin | 100,000 | | d-Amphetamine | 100,000 | | Atorvastatin | 20,000 | | Benzoylecgonine | 100,000 | | Buprenorphine | 15,000 | | Bupropion | 100,000 | | Caffeine | 100,000 | | Carbamazepine | 100,000 | | Cetirizine | 100,000 | | Chlorpheniramine | 100,000 | | Chlorpromazine | 50,000 | | Clomipramine | 100,000 | | Codeine | 100,000 | | Cyamemazine | 12,000 | | Desipramine | 100,000 | | Diphenhydramine | 100,000 | | Doxylamine | 100,000 | {9} | Interfering Substances | Maximum concetration tested without interference (ng/mL) | | --- | --- | | Duloxetin | 20,000 | | Fentanyl | 10,000 | | Fluoxetine | 100,000 | | Fluphenazine | 100,000 | | Gabapentin | 100,000 | | Hemoblogin | 300,000 | | Hydrocodone | 100,000 | | Hydromorphone | 100,000 | | Ibuprofen | 100,000 | | Imipramine | 100,000 | | Levomepromazine | 40,000 | | Lisinopril | 100,000 | | Losartan | 10,000 | | Loratidine | 100,000 | | MDA (3,4-methylene-dioxyamphetamine) | 100,000 | | MDEA | 100,000 | | MDMA (3,4-methylene-dioxymethylamphetamine) | 100,000 | | Meperidine | 100,000 | | Metformin | 100,000 | | Methylphenidate | 100,000 | | Metoprolol | 100,000 | | d-Methamphetamine | 100,000 | | Methapyrilene | 10,000 | | Methaqualone | 100,000 | | Metronidazole | 100,000 | | Morphine | 100,000 | | Nicotine | 100,000 | 10 {10} 11 | Interfering Substances | Maximun concentration tested without interference (ng/mL) | | --- | --- | | Nortriptyline | 100,000 | | Omeprazole | 100,000 | | Oxazepam | 100,000 | | Oxycodone | 100,000 | | Oxymorphone | 100,000 | | Phencyclidine | 10,000 | | Phenobarbital | 100,000 | | Promethazine | 5,000 | | (1S,2S)-(+) Pseudoephedrine | 100,000 | | Quetiapine | 100,000 | | Ranitidine | 100,000 | | Salbutamol (Albuterol) | 100,000 | | Sertraline | 5,000 | | THC-COOH (11-Nor-Delta-9-THC-9-carboxylic acid) | 1,000 | | Thioridazine | 20,000 | | L-Thyroxine | 10,000 | | Tramadol | 100,000 | | Verapamil | 100,000 | | Zolpidem | 10,000 | Structurally Unrelated Pharmacological Compounds for 300 ng/mL Cutoff: | Interfering Substances | Maximun concentration tested without interference (ng/mL) | | --- | --- | | Acetaminophen | 100,000 | | 6-Acetylmorphine | 10,000 | | Acetylsalicylic Acid | 100,000 | | Alimemazine | 4,000 | {11} | Interfering Substances | Maximun concetration tested without interference (ng/mL) | | --- | --- | | Amitriptyline | 100,000 | | Amlodipine | 100,000 | | Amoxicillin | 100,000 | | d-Amphetamine | 100,000 | | Atorvastatin | 20,000 | | Benzoylecgonine | 100,000 | | Buprenorphine | 15,000 | | Bupropion | 100,000 | | Caffeine | 100,000 | | Carbamazepine | 100,000 | | Cetirizine | 100,000 | | Chlorpheniramine | 100,000 | | Chlorpromazine | 100,000 | | Clomipramine | 100,000 | | Codeine | 100,000 | | Cyamemazine | 25,000 | | Desipramine | 100,000 | | Diphenhydramine | 100,000 | | Doxylamine | 100,000 | | Duloxetin | 60,000 | | Fentanyl | 10,000 | | Fluoxetine | 100,000 | | Fluphenazine | 100,000 | | Gabapentin | 100,000 | | Hemoglobin | 300,000 | | Hydrocodone | 100,000 | | Hydromorphone | 100,000 | | Ibuprofen | 100,000 | 12 {12} | Interfering Substances | Maximum concetration tested without interference (ng/mL) | | --- | --- | | Imipramine | 100,000 | | Levomepromazine | 100,000 | | Lisinopril | 100,000 | | Losartan | 10,000 | | Loratidine | 100,000 | | MDA (3,4-methylene-dioxyamphetamine) | 100,000 | | MDEA | 100,000 | | MDMA (3,4-methylene-dioxymethylamphetamine) | 100,000 | | Meperidine | 100,000 | | Metformin | 100,000 | | Methylphenidate | 100,000 | | Metoprolol | 100,000 | | d-Methamphetamine | 100,000 | | Methapyrilene | 40,000 | | Methaqualone | 100,000 | | Metronidazole | 100,000 | | Morphine | 100,000 | | Nicotine | 100,000 | | Nortriptyline | 100,000 | | Omeprazole | 100,000 | | Oxazepam | 100,000 | | Oxycodone | 100,000 | | Oxymorphone | 100,000 | | Phencyclidine | 20,000 | | Phenobarbital | 100,000 | | Promethazine | 15,000 | | (1S,2S)-(+) | 100,000 | | Pseudoephedrine | | 13 {13} | Interfering Substances | Maximun concetration tested without interference (ng/mL) | | --- | --- | | Quetiapine | 100,000 | | Ranitidine | 100,000 | | Salbutamol (Albuterol) | 100,000 | | Sertraline | 15,000 | | THC-COOH (11-Nor-Delta-9-THC-9-carboxylic acid) | 1,000 | | Thioridazine | 90,000 | | L-Thyroxine | 10,000 | | Tramadol | 100,000 | | Verapamil | 100,000 | | Zolpidem | 10,000 | ## Effect of Urine Specific Gravity To investigate the effect of urine specific gravity, urine samples with 1.004 to 1.025 specific gravity were spiked with EDDP at -25% of cut off and +25% of cut-off. Samples were then evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes. No interference was observed. ## Effect of pH Urine samples with pH 3 to 11 were spiked with EDDP at -25% of cut off and +25% of cut-off. Samples were then evaluated against the assay's calibration curve in both qualitative and semi-quantitative modes. No interference was observed. ## f. Assay cut-off: Characterization of how the device performs analytically around the claimed cutoff concentration is described in the precision section, M.1.a. above. ## 2. Comparison studies: a. Method comparison with predicate device: Method comparison studies for the LZI Methadone Metabolite Enzyme Immunoassay were conducted on the Beckman Coulter AU480 automated clinical analyzer. All samples were confirmed with LC/MS for both methadone and methadone metabolite {14} concentrations. For the 100 ng/mL cutoff, a total total of eighty-seven (87) unaltered clinical samples were tested. The results were identical when determined in qualitative or semi-quantitative mode, and are summarized in the following table: | Candidate Device Result | EDDP Concentration by LC/MS (ng/mL) | | | | | | --- | --- | --- | --- | --- | --- | | | Drug-free | Low Negative (<50% the cutoff conc) | Near Cutoff Negative (Between <50% below up to the cutoff conc) | Near Cutoff Positive (Between the cutoff and 50% above cutoff conc) | High Positive (>50% above the cutoff conc) | | Positive | 0 | 0 | 0 | 2 | 40 | | Negative | 23 | 11 | 9 | 2* | 0 | *Samples have discordant negative immunoassay results when compared to LC/MS, as summarized in the following table: | | Candidate Device Result | | EDDP LC/MS result (ng/mL) | | --- | --- | --- | --- | | Sample ID | Qualitative | Semi-quantitative | | | 45 | NEG | NEG | 103 | | 46 | NEG | NEG | 126 | For the 300 ng/mL cutoff, a total total of eighty-seven (84) unaltered clinical samples were tested. The results were identical when determined in qualitative or semi-quantitative mode, and are summarized in the following table: | Candidate Device Result | EDDP Concentration by LC/MS (ng/mL) | | | | | | --- | --- | --- | --- | --- | --- | | | Drug-free | Low Negative (<50% the cutoff conc) | Near Cutoff Negative (Between <50% below up to the cutoff conc) | Near Cutoff Positive (Between the cutoff and 50% above | High Positive (>50% above the cutoff conc) | {15} 16 | | | | | cutoff conc) | | | --- | --- | --- | --- | --- | --- | | Positive | 0 | 0 | 0 | 4 | 38 | | Negative | 21 | 15 | 6 | 0 | 0 | b. Matrix comparison: Not applicable 3. Clinical studies: a. Clinical Sensitivity: Not applicable b. Clinical specificity: No 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.