VITROS CHEMISTRY PRODUCTS THC REAGENT, CALIBRATOR KIT 30, FS CALIBRATOR 1, AND DAT PERFORMANCE VERIFIERS I, II, III, IV

{0} 1 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k063164 B. Purpose for Submission: New device C. Measurand: Cannabinoids in urine D. Type of Test: Homogenous enzyme immunoassay with associated calibrators and controls E. Applicant: Ortho Clinical Diagnostics, inc. F. Proprietary and Established Names: Vitros Chemistry Products THC Reagent VITROS Chemistry Products Calibrator Kit 30 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 862.3870 Cannabinoid test system 21 CFR 862.3200 Clinical toxicology calibrator 21 CFR 862.3280 Clinical toxicology control material 2. Classification: {1} Reagent and calibrators are Class II, control materials are class I 3. Product code: THC reagent: LDJ – Enzyme immunoassay, cannabinoids Calibrator kit 30: DLJ – Calibrators drug specific Calibrator 1: DKB – Calibrators drug mixture Control material: DIF – Drug mixture control materials 4. Panel: 91 - toxicology H. Intended Use: 1. Intended use(s): See indications for use below. 1. Indication(s) for use: VITROS Chemistry Products THC Reagent: For in vitro diagnostic use only. VITROS Chemistry Products THC Reagent is used on VITROS 5,1 FS Chemistry Systems for the semi-quantitative or qualitative determination of cannabinoids (THC) in human urine using a cutoff of either 20 ng/mL or 50 ng/mL. Measurements obtained with the VITROS THC method are used in the diagnosis and treatment of cannabinoid use or overdose. The VITROS Chemistry Products THC 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 30: For in vitro diagnostic use only. VITROS Chemistry Products Calibrator Kit 30 is used to calibrate VITROS 5,1 FS Chemistry Systems for the qualitative or semi-quantitative measurement of cannabinoids (THC). 2 {2} VITROS Chemistry Products FS Calibrator 1: For in vitro diagnostic use only. VITROS Chemistry Products FS Calibrator 1 is used in conjunction with VITROS Chemistry Products Calibrator Kits to calibrate VITROS 5,1 FS Chemistry Systems. VITROS Chemistry Products DAT Performance Verifiers I, II, III, IV and V: For in vitro diagnostic use only. 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 prescription use only 4. Special instrument requirements: VITROS 5,1 FS Chemistry System I. Device Description: The VITROS THC assay is a homogeneous enzyme immunoassay that is performed using the VITROS THC Reagent with the VITROS Calibrator Kit 30, VITROS FS Calibrator 1 and VITROS FS Diluent Pack 4 (DAT Diluent / DAT Diluent 2) on VITROS 5,1 FS Chemistry Systems. The VITROS THC Reagent is a dual chambered package containing ready-to-use liquid reagents that are used to detect cannabinoids in urine. Sample, calibrators, and controls are automatically treated with surfactant (DAT Diluent 2) prior to addition of reagents. Treated sample is added to Reagent 1 containing antibodies reactive to delta-9-tetrahydrocannabinol (delta-9-THC), glucose-6-phosphate and nicotinamide adenine dinucleotide (NAD+), followed by Reagent 2 containing delta-9-THC labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH). The assay is based on competition between delta 9-THC metabolites in the treated urine sample and the delta 9 -THC labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH) for antibody binding sites. Enzyme activity decreases upon binding to the antibody, therefore the concentration of delta-9-THC metabolites in the urine sample is directly proportional to measured enzyme activity. Active enzyme converts oxidized nicotinamide adenine dinucleotide (NAD+) to NADH, resulting in an absorbance change that is measured spectrophotometrically at 340 nm. VITROS Calibrator Kit 30 is prepared from human urine to which analyte, surfactant, and preservatives have been added. The VITROS FS Calibrator 1 is composed of processed water and 0.9% w/v sodium chloride (Saline). These calibrators are used to calibrate VITROS 5,1 FS Chemistry Systems for the qualitative or semiquantitative measurement of cannabinoids (THC). 3 {3} VITROS DAT Performance Verifiers I, II, III, IV and V are prepared from a human urine pool to which analytes, surfactant, and preservative have been added. These are assayed controls used to monitor performance of the VITROS THC assay on VITROS 5,1 FS Chemistry Systems. ## J. Substantial Equivalence Information: 1. Predicate device name(s): SYVA® EMIT® II Plus Cannabinoid Assay BIO-RAD® LiquichekTM Urine Toxicology Controls 2. Predicate 510(k) number(s): k011300, k022707 respectively 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | VITROS THC assay (device) | SYVA EMIT II Plus Cannabinoid assay and Liquichek Controls (predicate) | | Test Principle | Homogeneous enzyme immunoassay | Homogeneous enzyme immunoassay | | Sample Type | Human Urine | Human Urine | | Reagent Format | Liquid ready to use | Liquid ready to use | | Antibody | Monoclonal anti delta 9-tetrahydrocannabinol | Monoclonal anti delta 9-tetrahydrocannabinol | | Calibrator | 11-nor-delta-9-THC-9-COOH in human urine | 11-nor-delta-9-THC-9-COOH in human urine | | Calibrator and control format | Refrigerated: Liquid, ready to use | Refrigerated: Liquid, ready to use | | Control Matrix | Human urine | Human urine | | Differences | | | | --- | --- | --- | | Item | VITROS THC assay (device) | SYVA EMIT II Plus Cannabinoid assay and Liquichek Controls (predicate) | | Number of Calibrators | Six | Three for Qualitative Four for Semi-Quantitative | | Instrumentation | VITROS 5,1 FS Chemistry Systems | Multiple clinical chemistry systems | {4} | Differences | | | | --- | --- | --- | | Item | VITROS THC assay (device) | SYVA EMIT II Plus Cannabinoid assay and Liquichek Controls (predicate) | | | | | | Cutoff Values | 20 and 50 ng/mL | 20, 50, and 100 ng/mL | | Control claimed analytes | Cocaine metabolites (benzoylecgonine), benzodiazepines (lormetazepam), methadone, amphetamines (dmethamphetamine), Opiates (morphine), cannabinoids (11-nor-delta-9-THC-9-COOH), phencyclidine and barbiturates (secobarbital). | Methamphetamine, secobarbital, lormetazepam, Tetrahydrocannabinol(THC), benzoylecgonine, ethanol, lysergic acid diethylamide (LSD), methadone, methaqualone, morphine, (Free), phencyclidine, propoxyphene, nortriptyline and addition of creatinine, pH, specific gravity. | | Control: Number of levels | Five | Two | K. Standard/Guidance Document Referenced (if applicable): Not applicable L. Test Principle: Sample, calibrators, and controls are automatically treated with surfactant (DAT Diluent 2) prior to addition of reagents. Treated sample is added to Reagent 1 containing antibodies reactive to delta-9-tetrahydrocannabinol (delta-9-THC), glucose-6-phosphate and nicotinamide adenine dinucleotide $(\mathrm{NAD}+)$ , followed by Reagent 2 containing delta-9-THC labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH). The assay is based on competition between delta-9-THC metabolites in the treated urine sample and the delta-9 -THC labeled with the enzyme glucose-6-phosphate dehydrogenase (G6P-DH) for antibody binding sites. Enzyme activity decreases upon binding to the antibody, therefore the concentration of delta-9-THC metabolites in the urine sample is directly proportional to measured enzyme activity. Active enzyme converts oxidized nicotinamide adenine dinucleotide $(\mathrm{NAD}+)$ to NADH, resulting in an absorbance change that is measured spectrophotometrically at $340~\mathrm{nm}$ . M. Performance Characteristics (if/when applicable): {5} 6 1. Analytical performance: a. Precision/Reproducibility: The evaluation was conducted on the VITROS 5,1 FS Chemistry System on each of 22 calendar days using three lots of reagents on two different systems. Each run consisted of seven control fluids to challenge system precision at approximately ± 25% of the cut-off values (20 and 50 ng/mL), at the cut-off values, and at a high THC level (75 ng/mL). Control fluids are drug-spiked, pooled human urine with concentrations verified by GC/MS. Four calibrations were performed over the 22-day testing period. Multiple runs within a day were separated by at least two hours. All fluids were run in duplicate in each run. Final results and the calculation of within lab imprecision are based on weekly calibrations. The results of the test samples were analyzed by ANOVA as detailed in NCCLS EP5-A2. The mean within lab CV of the three lots tested was 3.5% (range 1.7 – 9.0%). Qualitative precision was estimated by determining the percentage of negative controls that produced a negative result versus the cutoff value and the percentage of positive controls that produced a positive result versus the cutoff value. | Cut-off Level | Cut-off | Number Tested | Correct Results | Confidence Level | | --- | --- | --- | --- | --- | | 20 ng/mL | 0.75x | 262 | 262 | >95% negative reading | | 20 ng/mL | 1.25x | 264 | 263 | >95% positive reading | | 50 ng/mL | 0.75x | 264 | 264 | >95% negative reading | | 50 ng/mL | 1.25x | 264 | 264 | >95% positive reading | One result pair (0.75x) 20ng/mL cutoff was excluded because the wrong fluid was tested. The results demonstrated acceptable imprecision on the VITROS THC Reagent lots and VITROS 5,1 FS Systems evaluated. b. Linearity/assay reportable range: The reportable range of the assay is from 5.0 to 80 ng/mL. A linearity study was performed according to NCCLS EP6-A. Urine pools containing low and high concentrations of 11-nor-delta-9-THC-9-COOH were mixed together resulting in 16 samples with 11-nor-delta-9-THC-9-COOH concentrations that spanned the measuring range of the assay. The linearity was evaluated with multiple reagent lots and assessed by comparing the measured VITROS THC assay values against the percent high pool to determine if the data conformed to a straight line. The linear regression calculations gave R² values of 0.999 and all samples were within the acceptance criteria when measured results {6} were compared to expected results across the sample range tested (2.5 – 82.5 ng/mL). c. Traceability, Stability, Expected values (controls, calibrators, or methods): The values assigned to the VITROS calibrators and controls for 11-nor-delta-9-THC-9-COOH are traceable to NIST standard SRM 1507b, verified by GC/MS. Working Calibrators that are value assigned by a GC/MS method with reference to NIST SRM 1507b are used to assign values to master calibrators using the Vitros 5,1 Chemistry system. These master calibrators are then used to assign values to final product calibrators. Controls are manufactured and verified by GC/MS traceable to NIST SRM 1507b. Final control values are assigned using the Vitros 5,1 Chemistry system. Real time stability studies are used to support both long term and open vial storage dates. d. Detection limit: The Limit of Blank (LOB), Limit of Detection (LOD) and Limit of Quantitation (LOQ) were determined following NCCLS EP17-A. The Limit of Blank, Limit of Detection and Limit of Quantitation were calculated for three reagent lots and determined to be 1.7 ng/mL, 4.9 ng/mL, and 5.0 ng/mL, respectively. The limit of quantitation is used when determining the reportable range of the assay. e. Analytical specificity: Determination of Cross-reactivity Cross-reactivity was examined by adding potential cross-reactant compounds into a drug free calibrator. The compounds were evaluated using the VITROS 5,1 FS Chemistry System to determine the VITROS THC Assay value equivalent to that of the calibration 11-nor-delta-9-THC-9-COOH analyte, at the cutoff values of 20 ng/mL and 50 ng/mL. Screened compounds exceeding the cutoff responses were further diluted with drug free calibrator to determine the approximate compound concentration required to generate a VITROS THC Assay value equivalent to 11-nor-delta-9-THC-9-COOH at the cutoff values. 7 {7} 8 # Cross Reacting Substances | Cross-reactant | Quantity (ng/mL) equivalent to 20 ng/mL of 11-nor-D9-THC-9-COOH | % Cross-reactivity* | Quantity (ng/mL) equivalent to 50 ng/mL of 11-nor-D9-THC-9-COOH | % Cross-reactivity* | | --- | --- | --- | --- | --- | | 8- *b*-11-dihydroxy-Δ⁹-THC | 24 | 83.3% | 60 | 83.3% | | Δ⁹-THC | 30 | 66.7% | 86 | 58.1% | | cannabinol | 36 | 55.6% | 121 | 41.3% | | 11-hydroxy-Δ⁹-THC | 38 | 52.6% | 105 | 47.6% | | cannabidiol | 6550 | 0.3% | 24500 | 0.2% | *The VITROS THC assay cutoff value (ng/mL) divided by the amount of cross-reactant (ng/mL) that produces a value equivalent to the cutoff value, multiplied by 100. # Exogenous Compounds Exogenous compounds were evaluated with a single test fluid spiked with the featured compound at a high concentration level (100,000 ng/mL). These compounds were spiked into commercially available calibrators with THC concentrations of 20 ng/mL or 50 ng/mL. Exogenous compounds demonstrating biases that exceeded the acceptance criteria were considered to interfere with the VITROS THC Assay at the cutoff value tested. When interference was observed, on-board dilutions of the test fluid were performed to estimate the highest interferent concentration that did not exceed the acceptance criteria at the cutoff value. | Compounds tested that do not interfere | | | | --- | --- | --- | | Compound | Concentration Tested Conventional | Concentration Tested SI | | ammonia | 570 mg/dL | 316 mmol/L | | amobarbitol | 10 mg/dL | 442 mmol/L | | ascorbic acid | 500 mg/dL | 28.4 mmol/L | | bilirubin | 26 mg/dL | 444 mmol/L | | brompheneramine | 10 mg/dL | 313 mmol/L | | calcium | 30 mg/dL | 7.5 mmol/L | | ciprofloxacin | 10 mg/dL | 302 mmol/L | | citric acid | 100 mg/dL | 5.2 mmol/L | | cloxacillin | 10 mg/dL | 230 mmol/L | {8} | Compounds tested that do not interfere | | | | --- | --- | --- | | Compound | Concentration Tested Conventional | Concentration Tested SI | | creatinine | 300 mg/dL | 26.5 mmol/L | | desipramine HCl | 10 mg/dL | 330 mmol/L | | dextromethorphan | 10 mg/dL | 369 mmol/L | | dicyclomine HCl | 10 mg/dL | 289 mmol/L | | diethylproprione | 10 mg/dL | 487 mmol/L | | doxylamine | 10 mg/dL | 370 mmol/L | | ethacrynic acid | 10 mg/dL | 330 mmol/L | | ethanol | 780 mg/dL | 169 mmol/L | | glucose | 4000 mg/dL | 222 mmol/L | | hemoglobin | 500 mg/dL | 5.00 g/L | | human IgG | 200 mg/dL | 2.00 g/L | | human serum albumin | 200 mg/dL | 2.00 g/L | | imipramine | 10 mg/dL | 357 mmol/L | | indomethacin | 10 mg/dL | 279 mmol/L | | iron | 0.1 mg/dL | 17.9 mmol/L | | L-hyoscyamine | 10 mg/dL | 346 mmol/L | | magnesium | 60 mg/dL | 24.7 mmol/L | | meperidine | 10 mg/dL | 404 mmol/L | | methoxyphenamine | 10 mg/dL | 558 mmol/L | | metronidazole | 10 mg/dL | 584 mmol/L | | nylidrine | 10 mg/dL | 334 mmol/L | | ofloxacin | 10 mg/dL | 277 mmol/L | | oxalic acid | 300 mg/dL | 23.8 mmol/L | | pH=9 | N/A | N/A | | phenylbutazone | 10 mg/dL | 324 mmol/L | | phenyltoloxamine | 10 mg/dL | 391 mmol/L | | phosphate | 950 mg/dL | 100 mmol /L | | potassium | 587 mg/dL | 150 mmol /L | | promethazine HCl | 10 mg/dL | 312 mmol/L | | propranolol | 10 mg/dL | 386 mmol/L | | pyruvate | 200 mg/dL | 22.8 mmol/L | | ranitidine | 10 mg/dL | 318 mmol/L | | riboflavin | 2 mg/dL | 53 mmol/L | | sodium | 6000 mg/dL | 2608 mmol/L | | tetrahydrozaline | 10 mg/dL | 499 mmol/L | | tolmetin/tolectin | 10 mg/dL | 390 mmol/L | | trihexylphenidyl HCl | 10 mg/dL | 296 mmol/L | {9} 10 | Compounds tested that do not interfere | | | | --- | --- | --- | | Compound | Concentration Tested Conventional | Concentration Tested SI | | trimethobenzamide | 10 mg/dL | 257 mmol/L | | tripelannamine | 10 mg/dL | 392 mmol/L | | tripolidine | 10 mg/dL | 359 mmol/L | | tyramine HCl | 10 mg/dL | 576 mmol/L | | urea | 3000 mg/dL | 500 mmol/L | | uric acid | 120 mg/dL | 7.1 mmol/L | ## Endogenous Compounds Solutions for the evaluation of potential endogenous interferents were prepared by addition of endogenous compounds a commercial calibrator containing THC. The test fluids and their control blanks were run on a VITROS 5,1 FS Chemistry. The effects of variations in pH were tested by adjusting the calibrator pH. Endogenous compounds demonstrating biases that exceeded the acceptance criteria were considered to interfere with the VITROS THC Assay for the appropriate cutoff value tested. When interference was observed, on-board dilutions of the test fluid were performed to estimate the highest interferent concentration that did not exceed the acceptance criteria at the cutoff value. ## Known Interfering Substances | Cutoff Value (ng/mL or μg/L) | Interferent | Interferent Concentration* | | Bias** (ng/mL or μg/L) | | --- | --- | --- | --- | --- | | | | Conventional | SI | | | 20 | pH | =4.0 | =4.0 | -3.9 | *The degree of interference at concentrations other than those listed might not be predictable from these results. Other interfering substances may be encountered in the patient population. **The bias is an estimate of the maximum difference observed. ## f. Assay cut-off: The cutoffs identified for this assay are 20 and 50 ng/mL. Performance around the cutoff was found to be acceptable and is shown in section 1.a., above. ## 2. Comparison studies: a. Method comparison with predicate device: A total of 113 unaltered human urine samples were assayed using the {10} VITROS THC Reagent, and the predicate device. Percent agreement was evaluated at each of the assay cutoff values (20 and 50 ng/mL). | | | Commercial Method | | | | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Cutoff Value | | Low Negative | Near Cutoff Negative | Near Cutoff Positive | High Positive | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 20 ng/mL | | (< -50%) < 10 ng/mL | (-50% to cutoff) 10-20 ng/mL | (cutoff to + 50%) 20-30 ng/mL | (> + 50%) > 30 ng/mL | 97.6% | 95.8% | 96.5% | | | VITROS positive | 0 | 1 | 15 | 53 | | | | | | VITROS negative | 30 | 11 | 3 | 0 | | | | | 50 ng/mL | | (< -50%) < 25 ng/mL | (-50% to cutoff) 25-50 ng/mL | (cutoff to + 50%) 50-75 ng/mL | (> + 50%) > 75 ng/mL | 100% | 87.0% | 94.7% | | | VITROS positive | 0 | 0 | 4 | 36 | | | | | | VITROS negative | 54 | 13 | 6 | 0 | | | | A total of 113 human urine samples were assayed using the VITROS THC Reagent and a GC/MS reference method specific for 11-nor-delta-9-THC-9-COOH. Percent agreement was evaluated at assay cutoff values of 20 and 50 ng/mL. | | | GC/MS Reference Method | | | | % Agreement | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | Cutoff Value | | Low Negative | Near Cutoff Negative | Near Cutoff Positive | High Positive | % Agreement Negative | % Agreement Positive | % Agreement Overall | | 20 ng/mL | | (< -50%) < 10 ng/mL | (-50% to cutoff) 10-20 ng/mL | (cutoff to + 50%) 20-30 ng/mL | (> + 50%) > 30 ng/mL | 62.9% | 100.0% | 77.0% | | | VITROS positive | 9 | 17 | 10 | 33 | | | | | | VITROS negative | 43 | 1 | 0 | 0 | | | | | 50 ng/mL | | (< -50%) < 25 ng/mL | (-50% to cutoff) 25-50 ng/mL | (cutoff to + 50%) 50-75 ng/mL | (> + 50%) > 75 ng/mL | 84.9% | 100.0% | 88.5% | | | VITROS positive | 5 | 8 | 11 | 16 | | | | | | VITROS negative | 70 | 3 | 0 | 0 | | | | {11} 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. 12