VITROS CHEMISTRY PRODUCTS AMPH REAGENT; CALIBRATOR KIT 26; FS CALIBRATOR 1; DAT PERFORMANCE VERIFIERS

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k062077 B. Purpose for Submission: Clearance of new device C. Measurand: Amphetamine D. Type of Test: Qualitative and semi-quantitative Enzyme Immunoassay E. Applicant: Ortho-Clinical Diagnostics, Inc. F. Proprietary and Established Names: VITROS Chemistry Products AMPH Reagent VITROS Chemistry Products Calibrator Kit 26 VITROS Chemistry Products FS Calibrator 1 VITROS Chemistry Products Dat Performance Verifiers I, II, III, IV and V G. Regulatory Information: 1. Regulation section: 21 CFR §862.3100, Amphetamine test system 21 CFR §862.3200, Clinical toxicology calibrators 21 CFR §862.3280, Clinical toxicology control material 2. Classification: Class II, II and Class I (reserved) {1} 3. Product code: DKZ, DLJ, DIF, respectively 4. Panel: Toxicology (91) H. Intended Use: 1. Intended use(s): See indications for use below. 2. Indication(s) for use: For in vitro diagnostic use only. VITROS Chemistry Products AMPH Reagent is used on VITROS 5,1 FS Chemistry Systems for the semi-quantitative and qualitative determination of amphetamines (AMPH) in human urine using a cutoff of either 500, or 1000 ng/mL. Measurements obtained with the VITRO AMPH method are used in the diagnosis and treatment of amphetamines use or overdose. The VITRO Chemistry Products AMPH assay is intended for use by professional laboratory personnel. It provides only a preliminary test result. A more specific alternative chemical method must be used to confirm a result obtained with this assay. Gas Chromatography/Mass Spectrometry (GC/MS) is the preferred confirmatory method. Clinical consideration and professional judgment should be applied to any drug-of-abuse test result, particularly when evaluating a preliminary positive result. VITROS Chemistry Products Calibrator Kit 26 is used to calibrate VITROS 5,1 FS Chemistry Systems for the qualitative and semi-quantitative measurement of drugs of abuse. VITROS Chemistry Products FS Calibrator 1 used in conjunction with VITROS Chemistry Products Calibrator Kits to calibrate VIROS 5,1 FS Chemistry Systems. VITROS Chemistry Products DAT Performance Verifiers are assayed controls used to monitor performance of urine drugs of abuse screening assays on VITROS 5,1 FS Chemistry Systems. 3. Special conditions for use statement(s): For use by professional laboratory personnel. For in vitro diagnostic use only. 2 {2} 4. Special instrument requirements: VITROS 5,1 FS Chemistry Systems (k031924) I. Device Description: The VITROS AMPH Reagent consists of a dual chambered package containing two liquid ready-to-use reagents used in a two-step reaction. Reagent 1 consists of murine monoclonal antibodies reactive to d-amphetamine and d-methamphetamine, NAD, and Glucose-6-phosphate. Reagent 2 consists of d-amphetamine and d-methamphetamine labeled with glucose-6-phosphate dehydrogenase. Calibrator Kit 26 is a standard that is sold separately. It is a one level of aqueous solution containing d-methamphetamine (AMPH), with a concentration of 2000 ng/mL. The standard is diluted with the FS Calibrator 1 to construct the standard curve which is used to calculate the concentration of the unknown samples. Verifiers I, II, III, IV and V are a set of 6 assayed controls (6 vials – 20 mL each) that are run with the samples to monitor the performance of the assay. They consist of human urine which drugs of abuse, metabolites of drugs of abuse, organic salt, surfactants and preservative have been added. J. Substantial Equivalence Information: 1. Predicate device name(s): Syva EMIT II Plus Amphetamines assay and Bio-Rad Liquicheck Urine Toxicology Controls 2. Predicate 510(k) number(s): k031004, k022707 respectively {3} 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | Device | Predicate | | Indications for Use | For in vitro diagnostic use only. The assay is intended for use in the qualitative and semi-quantitative analysis of amphetamines in human urine. The controls are assayed controls used to monitor the performance of Chemistry systems. | Same | | Test Principle | Homogeneous enzyme immunoassay | Same | | Specimen type | Human Urine | Same | | Reagent format | Liquid ready-to-use | Same | | Antibody source | Mouse monoclonal antibodies reactive to d-amphetamine and d-methamphetamine | Same | | Differences | | | | --- | --- | --- | | Item | Device | Predicate | | Calibrator levels | 6 levels | Qualitative – two levels Semi-quantitative – 300 ng/mL cutoff Value – four levels, 500 and 1000 ng/mL Cutoff values – five levels | | Calibrator format | Frozen liquid ready to use | Refrigerated liquid ready to use | | Control levels | Five | Two | | Control Analytes | Cocaine metabolites (benzoylecgonine), benzodiazepines (lormetazepam), methadone, amphetamines (d-methamphetamine), opiates (morphine), cannabinoids (11-nor-delta-THC-9-COOH), phencyclidine and barbiturates (secobarbital). | Methamphetamine, seco-barbital, lormetazepam, tetrahydrocannabinol (THC), benzoylecgonine, ethanol, lysergic acid diethylamide (LSD), methadone, methaqualone, morphine (FREE), phencyclidine, propoxyphene, nortriptyline and addition of creatinine, pH, specific gravity. | | Control Analytes cont. | | | {4} 5 K. Standard/Guidance Document Referenced (if applicable): CLSI EP5-A: Evaluation of Precision Performance of Clinical Chemistry Devices; Approved Guideline CLSI EP7-P; Interference Testing in Clinical Chemistry; Proposed Guideline CLSI EP6-P; Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Proposed Guideline CLSI EP12-A; User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline CLSI EP9-A2; Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline-Second Edition CLSI EP17-A; Protocols for Demonstration, Verification and Evaluation of Limits of Detection and Quantitation; Approved Guideline L. Test Principle: The test is an enzyme immunoassay for use on the Vitro 5,1 FS Chemistry System. Calibrators ranging in concentration from 0 to 2000 ng/mL are run with the assay. The Vitros AMPH assay is a homogenous enzyme immunoassay technique used for the qualitative and semi-quantitative analysis of amphetamine in human urine. In the performance of the Vitros AMPH assay, samples, calibrators and controls are treated with surfactant (DAT Diluent 2) prior to addition of the reagents. The treated sample is mixed with Reagent 1 which contains antibodies reactive to d-amphetamine and d-methamphetamine, glucose-6-phosphate and nicotinamide adenine dinucleotide (NAD+). Subsequently Reagent 2 containing d-amphetamine and d-methamphetamine, labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH) is added. Amphetamine in the treated sample and the d-amphetamine and d-methamphetamine labeled G6PDH compete for the antibody binding sites. Enzyme activity decreases upon binding to the antibody, so amphetamine concentration in the sample can be measured in terms of enzyme activity. Enzyme activity converts NAD+ to NADH resulting in an absorbance change that is measured spectrophotometrically at 340 nm. M. Performance Characteristics (if/when applicable): 1. Analytical performance: a. Precision/Reproducibility: The samples used for testing were the sponsor's controls, Verifiers I, II, III, IV and V. The samples were run in duplicate, twice a day for twenty-two days using two lots and four instruments. The results are presented in the table below: {5} | Control Mean Concentration (ng/mL) | Within Day SD | Within Lab SD | Within Lab CV% | | --- | --- | --- | --- | | 217 | 14.1 | 33.9 | 15.6 | | 370 | 12.1 | 23.6 | 6.4 | | 642 | 12.1 | 25.4 | 4.0 | | 784 | 11.8 | 28.1 | 3.6 | | 1255 | 27.1 | 71.6 | 5.7 | An additional study was performed using three quality control materials targeted at the assay's cutoff concentrations. The controls were run 20 replicates per day for five days using a single lot of reagent. The results are presented in the table below: | Cutoff Value (ng/mL) | Mean Measured Concentration | Within Lab SD | Within Lab CV% | | --- | --- | --- | --- | | 500 | 511 | 8.5 | 1.7 | | 1000 | 1041 | 15.1 | 1.5 | Qualitative imprecision was assessed by taking samples with target values $\pm 25\%$ of the cutoff concentration. The samples were run in duplicates one to two times a day for twenty-two days using one lot number of reagent and one analyzer. The results are in the table below: | Cutoff Value | Sample at ± 25% Cutoff | Number of observations | Number of correct Results | Confidence Level | | --- | --- | --- | --- | --- | | 500 ng/mL | 370 ng/mL | 84 | 84 | >95% negative reading | | | 642 ng/mL | 86 | 86 | >95% positive reading | | 1000 ng/mL | 784 ng/mL | 86 | 86 | >95% negative reading | | | 1255 ng/mL | 84 | 84 | >95% positive reading | # b. Linearity/assay reportable range: Linearity fluids were prepared from two pools of urine with amphetamine concentrations near the extremes of the calibration range (low pool $0\mathrm{ng / mL}$ and high pool $1800\mathrm{ng / mL}$ . The two pools were mixed to create 19 additional pools of intermediate concentrations. {6} Three determinations of each pool and three determinations of Verifiers were tested with three lots of reagent and one analyzer. A linear regression analysis was performed by the method of least squares. The plotted curve conforms to a straight line, supporting the reportable range 100-1450 ng/mL. Recovery study: Fourteen admixtures were prepared from two human urine pools and the concentrations were verified by GC/MS. Results are presented in the table below: | GC/MS ng/mL | VITROS AMPH Assay ng/mL | % Recovery | | --- | --- | --- | | 141 | 134 | 95.3 | | 211 | 216 | 102.3 | | 281 | 284 | 100.8 | | 422 | 412 | 97.7 | | 563 | 566 | 100.6 | | 704 | 751 | 106.8 | | 844 | 886 | 104.9 | | 985 | 1046 | 106.2 | | 1126 | 1137 | 101.0 | | 1196 | 1213 | 101.5 | | 1266 | 1267 | 100.1 | | 1301 | 1336 | 102.6 | | 1337 | 1349 | 100.9 | | 1407 | 1417 | 100.7 | c. Traceability, Stability, Expected values (controls, calibrators, or methods): A primary calibrator is prepared through gravimetric addition of USP (U.S. Pharmacopoeia) d-methamphetamine reference standard into drug free human urine and the concentration is confirmed by Gas Chromatography/Mass Spectrometry (GC/MS). Next working calibrators are prepared by diluting the primary calibrator with drug free human urine matrix. The value assigned to the working calibrator is verified by GC/MS. The VITROS DAT Performance Verifiers I,II,III,IV and V are prepared through the gravimetric addition of Sigma catalog number M8750 to drug free human urine matrix and the values are assigned by a GC/MS. Stability Studies: Real time and accelerated studies have been conducted. Protocols and acceptance criteria were described and found to be acceptable. The {7} manufacturer claims the following expiration date: When stored at 2-8 °C the assay reagent is good until the expiration date. When stored at ≤-18 °C the calibrator is good until the expiration date. Open stored at 2-8 °C it is good for ≤ 28 days. When stored at 2-8 °C the controls are good until the expiration date. Open stored at 2-8 °C they are good for 4 weeks. d. Detection limit: Seven human samples at various concentration below the lowest calibrator (150 ng/mL) were assayed in 10 replicates once a day for 5 days using three different lot numbers of reagent. A fully nested ANOVA was used to determine the total variability of each sample and a pooled SD was calculated for each of the three reagent lots used. The sponsor followed CLSI EP-17A to calculate the limit of quantitation which is 75 ng/mL for this assay. e. Analytical specificity: Cross-reactivity was established by spiking various concentrations of similarly structured compounds into drug-free calibrator. The quantity of a compound that produces a value equivalent to the d-methamphetamine (ng/mL) at each cutoff is listed below: | Compound | Quantity equivalent to 500 ng/mL cutoff | % Cross-reactivity | Quantity equivalent to 1000 ng/mL cutoff | % Cross-reactivity | | --- | --- | --- | --- | --- | | d-methamphetamine | 500 | 100 | 1,000 | 100 | | d-Amphetamine | 500 | 100 | 1,100 | 90.9 | | Benzphetamine | 513 | 97.5 | 1,032 | 96.9 | | l-Methamphetamine | 780 | 64.1 | 2,300 | 43.5 | | l-Amphetamine | 3000 | 16.7 | 10,000 | 10 | | Methylenedioxy-amphetamine | 1800 | 27.8 | 4,200 | 23.8 | | Methylenedioxy-ethylamine | 3700 | 13.5 | 17,500 | 5.7 | | Methylenedioxy-methamphetamine | 3400 | 14.7 | 20,500 | 4.9 | | 4-chloramphetamine | 4200 | 11.9 | 21,000 | 4.8 | | Mephentermine | 6300 | 7.9 | 45,000 | 2.2 | {8} | Compound | Quantity equivalent to 500 ng/mL cutoff | % Cross-reactivity | Quantity equivalent to 1000 ng/mL cutoff | % Cross-reactivity | | --- | --- | --- | --- | --- | | p-hydroxy-amphetamine | 10,000 | 5.0 | 65,000 | 1.5 | | Phentermine | 7500 | 6.7 | 35,000 | 2.9 | | Fenfluramine | 32,000 | 1.6 | >100,000 | <1.0 | | Methoxyphenamine | 100,000 | 0.5 | >100,000 | <1.0 | | Tranylcypromine | 32,000 | 1.6 | >100,000 | <1.0 | | Propanolol | 90,000 | 0.6 | >100,000, | <1.0 | | Tyramine | >100,000 | <0.5 | >100,000 | <1.0 | | Buproprion | >100,000 | <0.5 | >100,000 | <1.0 | | l-ephedrine | >100,000 | <0.5 | >100,000 | <1.0 | | d-ephedrine | >100,000 | <0.5 | >100,000 | <1.0 | | d-pseudoephedrine | >100,000 | <0.5 | >100,000 | <1.0 | | l-pseudoephedrine | >100,000 | <0.5 | >100,000 | <1.0 | | Nor-pseudoephedrine | >100,000 | <0.5 | >100,000 | <1.0 | | Phenylpropanolamine | >100,000 | <0.5 | >100,000 | <1.0 | | Chloroquine | >100,000 | <0.5 | >100,000 | <1.0 | | Phenothiazine | >100,000 | <0.5 | >100,000 | <1.0 | To evaluate interference the sponsor spiked potentially interfering compounds into drug-free calibrator. The compounds listed in the table below were found not to interfere according to the sponsor's criterion for bias $&lt; 95.6\mathrm{ng / mL}$ at $500\mathrm{ng / mL}$ and $&lt; 191\mathrm{ng / mL}$ at $1000\mathrm{ng / mL}$ amphetamine: | Compound | Concentration tested ng/mL | Compound | Concentration tested (ng/mL) | | --- | --- | --- | --- | | Albuterol | 100,000 | Meperidine | 100,000 | | Ammonia | 570 | Methylphenidate | 100,000 | | Ascorbic acid | 500 | Metronidazole | 100,000 | | Bilirubin | 26 | NaCl | 6000 | | Brompheniramine | 100,000 | Nylidrine | 100,000 | | Calcium | 30 | Ofloxacin | 100,000 | | Ciprofloxacin | 100,000 | Oxalic acid | 400 | | Citric acid | 100 | pH=4 | | | Cloxacillin | 100,000 | pH=9 | | | Creatinine | 300 | Phenothiazine | 100,000 | | Dextromethorphan | 100,000 | Phenyltoloxamine | 100,000 | | Dicyclomine | 100,000 | Phenylbutazone | 100,000 | {9} | Compound | Concentration tested ng/mL | Compound | Concentration tested (ng/mL) | | --- | --- | --- | --- | | Diethylproprione | 100,000 | Phosphate | 1420 | | Desipramine | 100,000 | Procainamide | 100,000 | | Doxylamine | 100,000 | Promethazine | 100,000 | | Ethacrynic acid | 100,000 | Pyruvate | 100 | | Ethanol | 780 | Quinacrine | 100,000 | | Glucose | 4000 | Ranitidine | 100,000 | | Hemoglobin | 500 | Riboflavin | 2 | | Human IgG | 200 | Setraline | 100,000 | | Human serum albumin | 200 | Tolmetin/tolectin | 100,000 | | Imipramine | 100,000 | Trihexylphenidyl | 100,000 | | Indomethacin | 100,000 | Trimethobenzamide | 100,000 | | Iron | 0.1 | Tripelannamine | 100,000 | | KCL | 1118 | Triprolidine | 100,000 | | l-hyoscyamine | 100,000 | Urea | 3000 | | Magnesium | 60 | Uric acid | 120 | Testing of high concentrations of NaCl, albumin and glucose showed no interference for a high specific gravity. f. Assay cut-off: Analytical performance of the device around the cutoff is described in Section1.M.d above. The test will yield a positive result when a given drug exceeds this concentration in the urine sample. 2. Comparison studies: a. Method comparison with predicate device: One hundred and six unaltered urine samples were assayed using the Vitros Chemistry products AMP reagent. The results were compared with the predicate device and Gas Chromatography/Mass Spectrometry (GC/MS) at the 500 ng/mL cutoff and the 1000 ng/mL cutoff. The results are presented below: {10} Comparison of Vitros AMP assay to the Predicate | | | Commercial Method | | | | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Cutoff Value | | Low Negative | Near Cutoff Negative | Near Cutoff Positive | High Positive | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 500 ng/mL | | (<50%) <250 ng/mL | (-50% to cutoff) 250-500 ng/mL | (cutoff to +50%) 500-750 ng/mL | (>+50%) >750 ng/mL | 96.3 | 98.1 | 97.2 | | | Vitros Positive | 0 | 2* | 5 | 46 | | | | | | Vitros negative | 36 | 16 | 1* | 0 | | | | | 1000 ng/mL | | (<50%) <500 ng/mL | (-50% to cutoff) 500-1000 ng/mL | (cutoff to +50%) 100-1500 ng/mL | (>+50%) >1500 ng/mL | 97.4 | 93.3 | 96.2 | | | Vitros Positive | 0 | 2* | 7 | 21 | | | | | | Vitros negative | 54 | 20 | 2* | 0 | | | | * See Summary of discordant results below: | Cutoff Value | Vitros AMPH Assay ng/mL | Commercial Method ng/mL | | --- | --- | --- | | 500 ng/mL | 491 | 542 | | | 521 | 401 | | | 739 | 477 | | 1000 ng/mL | 929 | 1232 | | | 998 | 1079 | | | 1021 | 947 | | | 1061 | 922 | {11} Comparison of Vitros AMP assay to the GC/MS | | | Commercial Method | | | | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Cutoff Value | | Low Negative | Near Cutoff Negative | Near Cutoff Positive | High Positive | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 500 ng/mL | | (<50%) <250 ng/mL | (-50% to cutoff) 250-500 ng/mL | (cutoff to +50%) 500-750 ng/mL | (>+50%) >750 ng/mL | 96.2 | 96.2 | 96.2 | | | Vitros Positive | 1* | 1* | 1 | 50 | | | | | | Vitros negative | 42 | 9 | 2* | 0 | | | | | 1000 ng/mL | | (<50%) <500 ng/mL | (-50% to cutoff) 500-1000 ng/mL | (cutoff to +50%) 100-1500 ng/mL | (>+50%) >1500ng/mL | 98.5 | 74.4 | 89.6 | | | Vitros Positive | 0 | 1* | 5 | 24 | | | | | | Vitros negative | 53 | 13 | 10* | 0 | | | | *See Summary of Discordant results below | Cutoff Value | Vitros AMPH Assay ng/mL | GC/MS ng/mL | Major Drug Identified by GC/MS | | --- | --- | --- | --- | | 500 ng/nL | 465 | 555 | amphetamine | | | 491 | 529 | amphetamine | | | 521 | 0 | none | | | 739 | 409 | methamphetamine | | 1000 ng/mL | 783 | 1009 | amphetamine | | | 816 | 1078 | amphetamine | | | 857 | 1048 | amphetamine | | | 861 | 1104 | amphetamine | | | 892 | 1139 | amphetamine | | | 893 | 1181 | amphetamine | | | 903 | 1061 | amphetamine | | | 921 | 1220 | amphetamine | | | 929 | 1144 | amphetamine | | | 998 | 1348 | amphetamine | | | 1303 | 912 | methamphetamine | {12} 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. 13