LZI COCAINE METABOLITE HOMOGENEOUS ENZYME IMMUNOASSAY,CALIBRATORS,CONTROLS

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k113139 B. Purpose for Submission: New device C. Measurand: Cocaine D. Type of Test: Qualitative and semi-quantitative enzyme immunoassay E. Applicant: Lin-Zhi International, Inc. F. Proprietary and Established Names: Cocaine Metabolite Enzyme Immunoassay Cocaine Metabolite Drugs of Abuse (DAU) Calibrators Cocaine Metabolite Drugs of Abuse (DAU) Controls G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | DIO – Cocaine and cocaine metabolite test system | II | 862.3250 | 91-Toxicology | | DLJ -Clinical toxicology calibrator | II | 862.3200 | 91- Toxicology | | LAS -Clinical toxicology control material | I, reserved | 862.3280 | 91- Toxicology | {1} H. Intended Use: 1. Intended use(s): See indications for use below 2. Indication(s) for use: The Cocaine Metabolite Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of benzoylecgonine in human urine, at a cutoff value of 150 ng/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 Chromatograph/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 result is positive. The Cocaine Metabolite Drugs of Abuse (DAU) Calibrators are for use as calibrators in the qualitative and semi-quantitative calibration of the Cocaine Metabolite Enzyme Immunoassay at a cutoff value of 150 ng/mL. The Cocaine Metabolite Drugs of Abuse (DAU) Controls are for use as assayed quality control materials to monitor the precision of the Cocaine Metabolite Enzyme Immunoassay at a cutoff value of 150 ng/mL. 3. Special conditions for use statement(s): For professional use only 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 testing was conducted on the Roche Diagnostics Hitachi 717 analyzer. 2 {2} I. Device Description: The assay consists of ready-to-use liquid reagents. Reagent 1 contains a mouse monoclonal anti-benzoylecgonine antibody, glucose-6-phosphate (G6P) nicotinamide adenine dinucleotide (NAD), stabilizers and sodium azide (0.09%) as a preservative. Reagent 2 contains glucose-6-phosphate dehydrogenase (G6PDH) labeled with benzoylecgonine in buffer with sodium azide (0.09%) as a preservative. The calibrators and controls are sold separately. The calibrator has 5 levels and the control has 2 levels. They consist of human urine samples containing benzoylecgonine with sodium azide (0.09%) as a preservative. J. Substantial Equivalence Information: 1. Predicate device name(s): Lin-Zhi International Inc., Cocaine Metabolite Enzyme Immunoassay Lin-Zhi International Inc., Cocaine Metabolite calibrators and controls 2. Predicate 510(k) number(s): k020763 and k020769, respectively 3. Comparison with predicate: | Similarities/Differences | | | | --- | --- | --- | | Item | Device | Predicates | | Intended Use | Intended for the qualitative and semi-quantitative determination of benzoylecgonine in human urine. The Calibrators are for use as calibrators in the qualitative and semi-quantitative calibration of the Cocaine Metabolite Enzyme Immunoassay. The Controls are for use as assayed quality control materials to monitor the precision of the Cocaine Metabolite Enzyme Immunoassay. | Same | | Analyte | Benzoylecgonine | Same | | Cutoff | 150 ng/mL | 300 ng/mL | | Matrix | Urine | Same | | Calibrators | 5 levels | 5 levels | {3} | Similarities/Differences | | | | --- | --- | --- | | Item | Device | Predicates | | | (0, 75, 150, 300, 1000 ng/mL) | (0, 150,300,1000, 3000 ng/mL) | | Controls | 2 levels (112.5 ng/mL, 187.5 ng/mL) | 2 levels (225 ng/mL, 375 ng/mL) | | Storage | 2-8 °C until expiration date | 2-8 °C until expiration date | ## K. Standard/Guidance Document Referenced (if applicable): CLSI EP5-A, Evaluation of Precision Performance of Clinical Chemistry Devices ## L. Test Principle: The Cocaine Metabolite assay is a homogenous enzyme immunoassay with ready-to-use liquid reagent. The assay is based on competition between drug in the sample and dug labeled with 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, benzoylecgonine-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 benzoylecgonine-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 nm. ## M. Performance Characteristics (if/when applicable): ### 1. Analytical performance: #### a. Precision/Reproducibility: Precision was determined by spiking benzoylecgonine into drug free urine at various concentrations (zero, -75%, -50%, -25%, at the cutoff, 125%, 150%, 175% and 200% of the cutoff). 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: | | | Within Run | | Between Run | | --- | --- | --- | --- | --- | | Sample concentration (ng/mL) | No. Observations | | No. Observations | | | | | # Neg/#Pos | | # Neg/#Pos | | 0 (negative) | 22 | 22/0 | 88 | 88/0 | | 37.5 (-75% c/o) | 22 | 22/0 | 88 | 88/0 | | 75 (-50% c/o) | 22 | 22/0 | 88 | 88/0 | | 112.5 (-25% c/o) | 22 | 22/0 | 88 | 88/0 | | 150 (cutoff) | 22 | 19/3 | 88 | 73/15 | | 187.5 (+25% c/o) | 22 | 0/22 | 88 | 0/88 | | 225 (+50% c/o) | 22 | 0/22 | 88 | 0/88 | | 267.5 (+50% c/o) | 22 | 0/22 | 88 | 0/88 | | 300 (+100% c/o) | 22 | 0/22 | 88 | 0/88 | Semi-Quantitative: | | | Within Run | | Between Run | | --- | --- | --- | --- | --- | | Sample concentration (ng/mL) | No. Observations | | No. Observations | | | | | # Neg/#Pos | | # Neg/#Pos | | 0 (negative) | 22 | 22/0 | 88 | 88/0 | | 37.5 (-75% c/o) | 22 | 22/0 | 88 | 88/0 | | 75 (-50% c/o) | 22 | 22/0 | 88 | 88/0 | | 112.5 (-25% c/o) | 22 | 22/0 | 88 | 88/0 | | 150 (cutoff) | 22 | 18/4 | 88 | 59/29 | | 187.5 (+25% c/o) | 22 | 0/22 | 88 | 0/88 | | 225 (+50% c/o) | 22 | 0/22 | 88 | 0/88 | | 267.5 (+50% c/o) | 22 | 0/22 | 88 | 0/88 | | 300 (+100% c/o) | 22 | 0/22 | 88 | 0/88 | b. Linearity/assay reportable range: Linearity across the range was confirmed by serially diluting a spiked urine pool containing $1000\mathrm{ng / mL}$ of benzoylecgonine in to concentration levels listed in the table below. Each sample was assayed 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: {5} | Expected Concentration (ng/mL) | Mean Observed Concentration (ng/mL) | Recovery (%) | | --- | --- | --- | | Negative | 4.0 | not applicable | | 48.1 | 58.1 | 120.8 | | 96.2 | 96.6 | 100.4 | | 144.3 | 151.4 | 104.9 | | 192.4 | 200.5 | 104.2 | | 288.6 | 312.1 | 108.1 | | 384.8 | 403.1 | 104.8 | | 481 | 488.3 | 101.5 | | 577.2 | 588.2 | 101.9 | | 673.4 | 694.8 | 103.2 | | 769.6 | 812 | 105.5 | | 865.8 | 917.5 | 106 | | 962 | 1017.8 | 105.8 | c. Traceability, Stability, Expected values (controls, calibrators, or methods): Five levels of calibrators (0, 75, 150, 300, 1000 ng/mL) and two levels of control material (112.5 ng/mL, 187.5 ng/mL) are available for use with the Cocaine Metabolite Enzyme Immunoassay. A commercially available benzoylecgonine standard solution from Cerilliant Analytical Reference Standards 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: Real time and accelerated 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 product is stable until expiration date which is 18 months. On board stability is good for 14 days when stored at 2-8 °C. d. Detection limit: Performance at low drug concentrations in the semi-quantitative assay was {6} characterized by determination of recovery (see section b above). # e. Analytical specificity: Cross-reactivity was established by spiking various concentrations of structurally related into drug-free urine. 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 are presented below: Structurally related | Compound | Equivalent to 150 ng/mL | % Cross reactivity | | --- | --- | --- | | Benzoylecgonine | 150 | 96.03% | | Cocaethylene | 4,000 | 4.58% | | Cocaine | 25,000 | 0.62% | | Ecgonine | 400,000 | 0.03% | | Ecgonine, Methyl Ester | 500,000 | 0% | | Norcocaine | 30,000 | 0.68% | | Atropine | 500,000 | 0% | Structurally unrelated | Compound | Tested concentrations ng/mL | -25% Benzoylecgonine | +25% Benzoylecgonine | | --- | --- | --- | --- | | Acetaminophen | 500,000 | Negative | Positive | | Acetylsalicylic Acid | 500,000 | Negative | Positive | | Amobarbital | 500,000 | Negative | Positive | | Amoxicillin | 500,000 | Negative | Positive | | Amphetamine | 500,000 | Negative | Positive | | Bupropion | 500,000 | Negative | Positive | | Catptopril | 500,000 | Negative | Positive | | Caffeine | 500,000 | Negative | Positive | | Chlordiazepoxide | 500,000 | Negative | Positive | | Chlorpheniramine | 500,000 | Negative | Positive | | Chlorpomazine | 500,000 | Negative | Positive | | Codeine | 500,000 | Negative | Positive | | Dextromethorphan | 500,000 | Negative | Positive | | Diazepam | 500,000 | Negative | Positive | | Digoxin | 500,000 | Negative | Positive | | Enalapril | 500,000 | Negative | Positive | | Fluoxetine | 100,000 | Negative | Positive | | Glyburide | 500,000 | Negative | Positive | | Ibuprofen | 500,000 | Negative | Positive | | Lidocaine | 500,000 | Negative | Positive | {7} | Compound | Tested concentrations ng/mL | -25% Benzoylecgonine | +25% Benzoylecgonine | | --- | --- | --- | --- | | Meperidine | 500,000 | Negative | Positive | | Methadone | 100,000 | Negative | Positive | | Methamphetamine | 500,000 | Negative | Positive | | Methaqualone | 500,000 | Negative | Positive | | Morphine | 500,000 | Negative | Positive | | Nicodine | 500,000 | Negative | Positive | | Nifedipine | 100,000 | Negative | Positive | | Oxazepam | 100,000 | Negative | Positive | | Phencyclidine | 500,000 | Negative | Positive | | Phenobarbital | 500,000 | Negative | Positive | | Propoxyphene | 100,000 | Negative | Positive | | Ranitidine | 500,000 | Negative | Positive | | Salicyluric acid | 500,000 | Negative | Positive | | Secobarbital | 500,000 | Negative | Positive | | 11-nor-THC-COOH | 500,000 | Negative | Positive | | Valproic Acid | 500,000 | Negative | Positive | | Verapamil | 500,000 | Negative | Positive | # Endogenous Compounds The following endogenous compounds were added into drug-free urine, urine sample spiked to $-25\%$ of benzoylecgonine and one urine spiked to $+25\%$ of benzoylecgonine at various concentrations. The substances listed in the table below were determined not to interfere at the concentration shown: | Interfering Substance | Concentration Tested mg/dL | -25% Benzoylecgonine | +25% Benzoylecgonine | | --- | --- | --- | --- | | Acetone | 1000 | Negative | Positive | | Ascorbic Acid | 400 | Negative | Positive | | Creatinine | 500 | Negative | Positive | | Ethanol | 1000 | Negative | Positive | | Galactose | 10 | Negative | Positive | | r-Globulin | 500 | Negative | Positive | | Glucose | 1500 | Negative | Positive | | Hemoglobin | 300 | Negative | Positive | | Human Serum Albumin | 500 | Negative | Positive | | Oxalic Acid | 100 | Negative | Positive | | Sodium Chloride | 3000 | Negative | Positive | | Urea | 2000 | Negative | Positive | {8} 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, 7, 8, 9, 10 and 11 were used. Each of these samples were divided into two aliquots for each drug and spiked to -25% of the cutoff and +25% of the cutoff. No positive or negative interference due to pH was observed. To test for possible positive and/or negative interference from specific gravity urine samples having specific gravity from 1.000, 1.002, 1.005, 1.007, 1.010, 1.015, 1.017, 1.025 and 1.030 were used. Each of these samples were divided into two aliquots for each drug and 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: Eighty unaltered clinical urine samples (40 negative and 40 positive) were evaluated by the Lin-Zhi Cocaine Metabolite Enzyme Immunoassay and compared to LC/MS. Results from the study are presented below: Qualitative | Candidate Device Results | Negative | Less than half the cutoff concentration by GC/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 | 2 | 5 | 31 | | Negative | 20 | 4 | 14 | 4 | 0 | % Agreement among positives is 90% (36/40) % Agreement among negatives is 95% (38/40) {9} 10 Discordant | Cutoff Value (ng/mL) | LZI Cocaine Assay (POS/NEG) | Drug/Metabolite LC/MS value (ng/mL) | | --- | --- | --- | | 150 | Positive | 141 | | 150 | Positive | 147 | | 150 | Negative | 150 | | 150 | Negative | 152 | | 150 | Negative | 167 | | 150 | Negative | 176 | Semi-quantitative | Candidate Device Results | Negative | Less than half the cutoff concentration by GC/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 | 1 | 5 | 31 | | Negative | 20 | 4 | 15 | 4 | 0 | % Agreement among positives is 90% (36/40) % Agreement among negatives is 98% (39/40) Discordant | Cutoff Value (ng/mL) | LZI 6-Acetylmorphine Assay (POS/NEG) | Drug/Metabolite LC/MS value (ng/mL) | | --- | --- | --- | | 150 | Positive | 141 | | 150 | Negative | 150 | | 150 | Negative | 167 | | 150 | Negative | 167 | | 150 | Negative | 176 | b. Matrix comparison: Test is for urine samples only 3. Clinical studies: a. Clinical Sensitivity: Not applicable b. Clinical specificity: {10} 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. 11