ARK Oxcarbazepine Metabolite Assay, ARK Oxcarbazepine Metabolite Calibrator, ARK Oxcarbazepine Metabolite Control

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k153596 B. Purpose for Submission: New Device C. Measurand: Oxcarbazepine Metabolite D. Type of Test: Homogenous enzyme immunoassay E. Applicant: ARK Diagnostics, Inc. F. Proprietary and Established Names: ARK Oxcarbazepine Metabolite Assay G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | POX | II | 862.3645 - Neuroleptic drugs radioreceptor assay test system. | 91 - Toxicology | | DLJ | II | 862.3200 - Clinical toxicology calibrator | 91 - Toxicology | | LAS | I, reserved | 862.3280 - Clinical toxicology control material | 91 - Toxicology | H. Intended Use: 1. Intended use(s): See indications for use. {1} 2. Indication(s) for use: The ARK Oxcarbazepine Metabolite Assay is a homogeneous enzyme immunoassay intended for the quantitative determination of Oxcarbazepine Metabolite in human serum on automated clinical chemistry analyzers. The measurements obtained are used in monitoring levels of Oxcarbazepine Metabolite to help ensure appropriate therapy. The ARK Oxcarbazepine Metabolite Calibrator is intended for use in calibration of the ARK Oxcarbazepine Metabolite Assay. The ARK Oxcarbazepine Metabolite Control is an assayed quality control material intended for use in quality control of the ARK Oxcarbazepine Metabolite Assay. 3. Special conditions for use statement(s): For prescription use only 4. Special instrument requirements: The assay was validated on the Beckman Coulter AU480 automated clinical chemistry analyzer. I. Device Description: The ARK Oxcarbazepine Metabolite Assay is a homogeneous immunoassay based on competition between drug in the specimen and Oxcarbazepine Metabolite labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for binding to the antibody reagent. The ARK Oxcarbazepine Metabolite Assay consists of reagents R1 (anti-Oxcarbazepine Metabolite polyclonal antibody with substrate) and R2 (Oxcarbazepine Metabolite labeled with bacterial G6PDH enzyme). ARK Oxcarbazepine Metabolite Calibrator is comprised of a synthetic protein matrix and consists of a six-level set to calibrate the assay. The ARK Oxcarbazepine Metabolite Control is comprised of a synthetic protein matrix and consists of a three level set used for quality control of the assay. J. Substantial Equivalence Information: 1. Predicate device name(s): Emit 2000 Carbamazepine Assay, ARK Topiramate Calibrator, ARK Topiramate Control 2. Predicate 510(k) number(s): k010814 (assay); k083799 (calibrator and control) {2} 3. Comparison with predicate: | Assay Similarities | | | | --- | --- | --- | | Item | Candidate Device (ARK Oxcarbazepine Assay) | Predicate Device (Emit 2000 Carbamazepine Assay, k010814) | | Intended Use/Indications for Use | The ARK Oxcarbazepine Metabolite Assay is intended for the quantitative determination of Oxcarbazepine Metabolite in human serum on automated clinical chemistry analyzers. The measurements obtained are used in monitoring levels of Oxcarbazepine Metabolite to help ensure appropriate therapy. | The Emit® 2000 Carbamazepine Assay is a homogeneous enzyme immunoassay intended for use in the quantitative analysis of carbamazepine in human serum or plasma. The results obtained helps physicians individualize dosage regimens. | | Sample Type | Serum | Serum or plasma | | Methodology | Homogenous enzyme immunoassay (EIA) | Same | | Reagent components | Two (2) reagent system: Anti-Oxcarbazepine Metabolite Antibody/Substrate Reagent (R1) containing rabbit polyclonal antibodies to Oxcarbazepine Metabolite, glucose-6-phosphate, nicotinamide adenine dinucleotide, bovine serum albumin, sodium azide, and stabilizersEnzyme Reagent (R2) containing Oxcarbazepine Metabolite labeled with bacterial G6PDH, buffer, bovine serum albumin, sodium azide, and stabilizers | Two (2) reagent system: Antibody/Substrate Reagent (R1) containing mouse monoclonal antibodies to carbamazepine, glucose-6-phosphate, nicotinamide adenine dinucleotide.Enzyme Reagent (R2) containing carbamazepine labeled with bacterial G6PDH, buffer.Sodium azide, buffer, preservatives, and stabilizers | | Testing environment | Routine clinical laboratory | Same | | Reagent condition and | Liquid, 2-8°C | Same | {3} 4 | Assay Similarities | | | | --- | --- | --- | | Item | Candidate Device (ARK Oxcarbazepine Assay) | Predicate Device (Emit 2000 Carbamazepine Assay, k010814) | | storage | | | | Assay Differences | | | | --- | --- | --- | | Item | Candidate Device (ARK Oxcarbazepine Assay) | Predicate Device (ARK Topiramate Assay, k083799) | | Analyte | Oxcarbazepine metabolite | Carbamazepine | | Controls: Similarities and Differences | | | | --- | --- | --- | | Item | Candidate Device (ARK Oxcarbazepine Control) | Predicate Device (ARK Topiramate Control; k083799) | | Intended Use/Indications for Use | The ARK Oxcarbazepine Metabolite control is an assayed quality control material intended for use in quality control of the ARK Oxcarbazepine Metabolite Assay. | The ARK Topiramate Control is intended for use in quality control of the ARK Topiramate Assay. | | Matrix | Synthetic protein matrix (buffer, bovine serum albumin, preservatives) | Same | | Levels | 3 (LOW, MID, HIGH) | Same | | Calibrators: Similarities and Differences | | | | --- | --- | --- | | Item | Candidate Device (ARK Oxcarbazepine Calibrator) | Predicate Device (ARK Topiramate Calibrator; k083799) | | Intended Use/Indications for Use | ARK Oxcarbazepine Metabolite Calibrator is intended for use in calibration of the ARK Oxcarbazepine Metabolite Assay. | The ARK Topiramate Calibrator is intended for use in calibration of the ARK Topiramate Assay. | | Matrix | Synthetic protein matrix (buffer, bovine serum albumin, preservatives) | Same | | Levels | 6 | Same | {4} K. Standard/Guidance Document Referenced (if applicable): - Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline- Third Edition (EP05-A3) - Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition (EP07-A2) - Measurement Procedure Comparison and Bias Estimation Using Patient Samples; Approved Guideline - Third Edition (EP09-A3) - Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline (EP6-A) - Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second Edition (EP17-A2) L. Test Principle: The ARK Oxcarbazepine Metabolite Assay is a homogeneous immunoassay based on competition between drug in the specimen and Oxcarbazepine Metabolite labeled with the enzyme glucose-6-phosphate dehydrogenase (G6PDH) for binding to the antibody reagent. As the latter binds antibody, enzyme activity decreases. In the presence of drug from the specimen, enzyme activity increases and is directly proportional to the drug concentration. Active enzyme converts the coenzyme nicotinamide adenine dinucleotide (NAD) to NADH that is measured spectrophotometrically as a rate of change in absorbance. Endogenous serum G6PDH does not interfere with the results because the coenzyme NAD functions only with the bacterial enzyme used in the assay. M. Performance Characteristics (if/when applicable): 1. Analytical performance: a. Precision/Reproducibility: Data for the precision evaluation studies (total and within-laboratory precision) were collected on a single Beckman Coulter AU480 automated clinical chemistry analyzer. Each level of control and patient sample was assayed in quadruplicate twice a day over twenty non-consecutive days. A total of 160 determinations were made for each sample. One calibration was performed during this interval. A total of 6 samples (3 levels of human serum pooled specimens containing Oxcarbazepine Metabolite, and 3 levels of ARK Oxcarbazepine Metabolite controls) were used in the study. Mean oxcarbazepine metabolite, standard deviation (SD) and coefficients of variation (%CVs) were calculated for within-run, between-day, and total precision. 5 {5} | Sample | N | Mean (g/mL) | Within Run | | Between Day | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | | ARK Control | | | | | | | | | | LOW | 160 | 3.0 | 0.12 | 4.0 | 0.12 | 4.1 | 0.17 | 5.7 | | MID | 160 | 10.1 | 0.37 | 3.6 | 0.33 | 3.2 | 0.48 | 4.8 | | HIGH | 160 | 30.2 | 0.99 | 3.3 | 1.19 | 3.9 | 1.54 | 5.1 | | Human Serum | | | | | | | | | | LOW | 160 | 3.1 | 0.12 | 3.9 | 0.12 | 4.0 | 0.17 | 5.5 | | MID | 160 | 10.1 | 0.38 | 3.8 | 0.36 | 3.6 | 0.55 | 5.5 | | HIGH | 160 | 30.4 | 1.10 | 3.6 | 1.11 | 3.7 | 1.55 | 5.1 | # b. Linearity/assay reportable range: Linearity was evaluated according to the CLSI EP6-A standard. A series of 8 concentrations (1.00, 3.00, 5.00, 10.00, 20.00, 30.00, 40.00, and 50.00 g/mL) in human serum were prepared by proportionally diluting a spiked high Oxcarbazepine Metabolite concentration serum sample pool with a negative serum pool. Two analytical runs of three replicates of each sample (6 replicates total) were measured for each concentration, and the mean of the measured concentration was compared to the theoretical expected Oxcarbazepine Metabolite concentrations based on the first order and second order linear regression analysis. The linear regression results using the sample concentrations listed above are: | Analyzer | Slope | Intercept | R2 | | --- | --- | --- | --- | | AU480 | 1.0388 | -0.0693 | 0.9994 | This evaluation supports linearity within the claimed measuring range of 1.0 to 37.0 g/mL. # Recovery An analytical recovery study was conducted on the Beckman Coulter AU480 to determine the effect of differing ratios of Oxcarbazepine Metabolite enantiomers (S- or R- enantiomer; ratio hereafter S:R) on recovery by spiking Oxcarbazepine Metabolite into human serum negative for Oxcarbazepine Metabolite. The results are as follows: {6} | | Mean Recovered Concentration (g/mL) | | | | | --- | --- | --- | --- | --- | | Theoretical Concentration (g/mL) | S:R 1:1 | S:R 4:1 | S:R 9:1 | S:R 19:1 | | 1.0 | 0.77 | 0.93 | 0.98 | 0.95 | | 4.0 | 3.78 | 3.92 | 3.94 | 3.86 | | 8.0 | 7.47 | 8.18 | 8.16 | 7.82 | | 15.0 | 14.10 | 15.80 | 14.91 | 15.42 | | 20.0 | 19.03 | 21.69 | 19.81 | 21.02 | | 35.0 | 33.74 | 34.71 | 33.52 | 36.16 | | 45.0 | 42.89 | 46.88 | 44.63 | 49.46 | c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability and Value Assignment: ARK Oxcarbazepine Metabolite Calibrators and ARK Oxcarbazepine Metabolite Controls are traceable to a commercial product, and are value assigned by gravimetric dilution. Stability: Shelf life: Accelerated and real-time shelf-life stability study protocols and acceptance criteria were reviewed and found to be adequate. Accelerated stability studies were performed to support a shelf-life stability claim until the expiration date for the ARK Oxcarbazepine Metabolite Calibrators and Controls when stored unopened at $2 - 8^{\circ}\mathrm{C}$ . Real time stability studies to support shelf-life stability claims are ongoing. Open vial: The Sponsor's open vial stability study protocols and acceptance criteria were reviewed and found to be adequate. Real-time testing for opened vial ARK Oxcarbazepine Metabolite Calibrators and Controls stability was performed and supports stability claims of up to 12 months when stored opened at $2 - 8^{\circ}\mathrm{C}$ . d. Detection limit: The LoB was determined using pooled human serum negative for oxcarbazepine metabolite as the blank sample. A single concentration of oxcarbazepine metabolite $(0.3\mathrm{g / mL})$ in serum was tested to assess LoD. To determine the LoB and LoD, twenty (20) replicates of each sample were tested in each of three (3) runs to yield sixty (60) replicates of each sample tested, in a single day. The grand mean, standard deviation, and coefficient of variation for each test sample were calculated to determine inter-assay precision. The LOQ of the ARK Oxcarbazepine Metabolite Assay was determined as the lowest concentration for which acceptable inter-assay precision and recovery is observed {7} $(\leq 20\% \mathrm{CV}$ with $\pm 15\%$ recovery). Multiple low concentration Oxcarbazepine Metabolite serum samples were prepared gravimetrically. Eight (8) replicates of each sample were tested in each of five (5) runs to give a minimum of 40 replicates of each LOQ sample tested. The grand mean, standard deviation, and coefficient of variation for each test sample were calculated to determine inter-assay precision. The sponsor determined the LoB, LoD, and LoQ to be the following: | Criterion | Oxcarbazepine Metabolite Concentration (g/mL) | | --- | --- | | Limit of Blank (LoB): N = 60 | 0.01 | | Limit of Detection (LoD); N = 60 | 0.06 | | Limit of Quantitation (LoQ); N=40 | 1.0 | # e. Analytical specificity: Specificity studies were conducted for metabolites of Oxcarbazepine and structurally related compounds (including Oxcarbazepine itself), medications routinely coadministered with Oxcarbazepine, other over-the-counter drugs, and endogenous substances, where Oxcarbazepine Metabolite $(20\mu \mathrm{g} / \mathrm{mL})$ was present in serum. Concentrations tested and results are tabulated in the sections below. Oxcarbazepine Metabolites: | Metabolite | Level Tested (μg/mL) | % Cross-Reactivity | % Interference | | --- | --- | --- | --- | | MHD Glucuronide | 20 | 1.6 | 1.6 | | | 40 | 0.0 | -0.1 | | | 100 | 1.5 | 7.4 | | | 200 | 1.0 | 10.5 | | Dihydro-carbamazepine | 5.0 | 6.0 | 1.5 | | Carbamazepine-epoxide | 10.0 | 13.6 | 6.9 | | Dihydro-dihydroxy carbamazepine (DHD) | 5.0 | -11.3 | -2.9 | | Oxcarbazepine | 20.0 | 22.2 | 22.6 | | Carbamazepine | 20.0 | 20.4 | 20.7 | | Eslicarbazepine acetate | 20.0 | 22.1 | 22.4 | {8} 9 # Endogenous Substances: Endogenous substances were tested at 3 and 30 g/mL Oxcarbazepine Metabolite in serum. No significant interference was observed. Concentrations tested and results are shown below. No significant interference was observed. | | | Percentage recovery | | | --- | --- | --- | --- | | Interfering Substance | Interferent Concentration | 3 μg/mL Oxcarbazepine Metabolite | 30 μg/mL Oxcarbazepine Metabolite | | Human Albumin | 12 g/dL | 102.2 | 95.1 | | Bilirubin (conjugated) | 70 mg/dL | 108.6 | 100.2 | | Bilirubin (unconjugated) | 70 mg/dL | 102.7 | 92.4 | | Cholesterol | 602 mg/dL | 96.5 | 103.5 | | Human IgG | 12 g/dL | 105.7 | 100.7 | | Hemoglobin | 1000 mg/dL | 101.0 | 103.9 | | Rheumatoid Factor | 1000 IU/mL | 93.1 | 93.1 | | Triglycerides | 1000 mg/dL | 96.6 | 94.3 | | Uric Acid | 30 mg/dL | 107.5 | 95.5 | # Co-administered drugs and common OTC drugs: The sponsor evaluated the effect of co-administered drugs and common over the counter drugs on the measurement of Oxcarbazepine Metabolite in pooled human serum containing either low (3.0 μg/mL) or high (30.0 μg/mL) levels of Oxcarbazepine Metabolite. No significant interference was observed. | # | Compound | Concentration Tested | Percentage Recovery | | | --- | --- | --- | --- | --- | | | | | Oxcarbazepine metabolite (3 g/mL) | Oxcarbazepine metabolite (30 g/mL) | | 1 | Acetaminophen | 200 | 95.6 | 97.1 | | 2 | Acetazolamide | 100 | 99.9 | 90.3 | | 3 | Acetylsalicylic acid | 100 | 95.1 | 96.0 | | 4 | Amikacin | 100 | 91.7 | 92.0 | | 5 | Amitriptyline | 10 | 105.1 | 101.1 | | 6 | Amoxapine | 10 | 99.3 | 98.0 | | 7 | Amphotericin B | 100 | 93.6 | 93.2 | | 8 | Ampicillin | 100 | 96.5 | 100.2 | {9} | # | Compound | Concentration Tested | Percentage Recovery | | | --- | --- | --- | --- | --- | | | | | Oxcarbazepine metabolite ( 3 g/mL) | Oxcarbazepine metabolite (30 g/mL) | | 9 | Ascorbic acid | 100 | 92.8 | 91.1 | | 10 | Baclofen | 100 | 91.1 | 93.5 | | 11 | Bupropion | 10 | 109.6 | 98.8 | | 12 | Caffeine | 100 | 98.3 | 91.7 | | 13 | Chloramphenicol | 250 | 93.7 | 90.3 | | 14 | Chlorpromazine | 10 | 98.3 | 99.7 | | 15 | Citalopram | 10 | 102.9 | 99.3 | | 16 | Clobazam | 100 | 98.3 | 103.2 | | 17 | Clonazepam | 10 | 104.6 | 99.2 | | 18 | Cyclosporin A | 40 | 91.2 | 90.2 | | 19 | Diazepam | 20 | 103.1 | 100.3 | | 20 | Digoxin | 10 | 97.3 | 97.0 | | 21 | Doxepin | 10 | 107.4 | 102.9 | | 22 | Erythromycin | 200 | 94.5 | 94.7 | | 23 | Ethanol | 4000 (0.4%) | 91.6 | 100.7 | | 24 | Ethotoin | 100 | 98.4 | 96.2 | | 25 | Ethosuximide | 250 | 103.2 | 105.1 | | 26 | Felbamate | 250 | 93 | 93.8 | | 27 | Fluoxetine | 20 | 94.9 | 99.2 | | 28 | Furosemide | 100 | 95.2 | 92.8 | | 29 | Gabapentin | 200 | 92.2 | 104.3 | | 30 | Gentamicin | 100 | 95.8 | 91.2 | | 31 | Haloperidol | 10 | 101.2 | 97.4 | | 32 | Heparin | 200 U/mL | 96.3 | 92.3 | | 33 | Ibuprofen | 500 | 103.3 | 91.6 | | 34 | Imipramine | 10 | 109.4 | 100.4 | | 35 | Kanamycin A | 200 | 93.8 | 109.0 | | 36 | Lamotrigine | 400 | 91.5 | 97.9 | | 37 | Levetiracetam | 400 | 97.7 | 94.7 | | 38 | Lidocaine | 100 | 96.8 | 97.7 | | 39 | Lincomycin | 1000 | 90.7 | 100.4 | | 40 | Mephenytoin | 100 | 100.7 | 97.3 | {10} | # | Compound | Concentration Tested | Percentage Recovery | | | --- | --- | --- | --- | --- | | | | | Oxcarbazepine metabolite ( 3 g/mL) | Oxcarbazepine metabolite (30 g/mL) | | 41 | Mesoridazine | 10 | 97.8 | 99.4 | | 42 | Methicillin | 250 | 93.5 | 96.2 | | 43 | Naproxen | 600 | 102.2 | 95.7 | | 44 | Neomycin | 100 | 95.6 | 102.9 | | 45 | Niacin | 100 | 93 | 93.9 | | 46 | Nitrazepam | 20 | 106.3 | 98.5 | | 47 | Notriptyline | 10 | 104.4 | 102.0 | | 48 | Olanzapine | 10 | 105.8 | 100.5 | | 49 | Paroxetine | 10 | 96.7 | 98.3 | | 50 | 2-phenyl-2-ethyl- malonamide (PEMA) | 100 | 94.6 | 93.9 | | 51 | Penicillin V | 100 | 95.4 | 93.8 | | 52 | Perphanazine | 50 | 104.9 | 100.9 | | 53 | Phenobarbital | 200 | 90.2 | 94.7 | | 54 | Phenytoin | 200 | 100.1 | 99.6 | | 55 | Pregabalin | 200 | 91.5 | 90.2 | | 56 | Primidone | 100 | 95 | 92.4 | | 57 | Procainamide | 100 | 93.3 | 92.4 | | 58 | Prochlorperazine | 10 | 105.2 | 101.6 | | 59 | Ranitidine | 100 | 102.1 | 100.6 | | 60 | Rifampin | 100 | 93.3 | 92.7 | | 61 | Risperidone | 10 | 100.6 | 97.7 | | 62 | Sertraline | 100 | 98.9 | 93.4 | | 63 | Spectinomycin | 100 | 97.2 | 97.9 | | 64 | Stiripentol | 100 | 93.8 | 99.7 | | 65 | Sulfamethoxazole | 400 | 100.5 | 97.5 | | 66 | Theophylline | 200 | 100.5 | 100.8 | | 67 | Thioridazine | 10 | 103.9 | 98.0 | | 68 | Tobramycin | 100 | 94.5 | 101.3 | | 69 | Tiagabine | 200 | 91.6 | 93.5 | | 70 | Topiramate | 250 | 92.8 | 91.7 | | 71 | Trimethoprim | 40 | 101.2 | 93.6 | | 72 | Valproic Acid | 600 | 92.7 | 93.0 | {11} 12 | # | Compound | Concentration Tested | Percentage Recovery | | | --- | --- | --- | --- | --- | | | | | Oxcarbazepine metabolite (3 g/mL) | Oxcarbazepine metabolite (30 g/mL) | | 73 | Vancomycin | 250 | 101.3 | 92.6 | | 74 | Vigabatrin | 150 | 103.2 | 96.9 | | 75 | Zonisamide | 400 | 92.1 | 91.4 | f. Assay cut-off: Not applicable 2. Comparison studies: a. Method comparison with predicate device: Method comparison studies were performed according to the CLSI EP09-A3. One-hundred-Ninety (190) individual human serum samples ranging in concentration from 1.7 g/mL to 36.4 g/mL were measured using the ARK Oxcarbazepine Metabolite Assay and compared to the results obtained using LC-MS/MS. Results of the Passing Bablock regression analysis are presented below. | Analyzer | Slope (95% CI) | Intercept (95% CI) | N | R² (95% CI) | Sample Range Tested | | --- | --- | --- | --- | --- | --- | | AU480 | 1.01 (0.96 to 1.04) | -0.38 (-0.84 to 0.12) | 190 | 0.95 (0.94 to 0.97) | 1.7-36.4 g/mL | b. Matrix comparison: Not applicable. 3. Clinical studies: a. Clinical Sensitivity: Not applicable b. Clinical specificity: Not applicable {12} 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: The package insert includes the following statement: "A reference range for TDM of Oxcarbazepine Metabolite (MHD) has not been well established. A wide range of MHD serum concentrations (3-35 µg/mL) have been observed (established by reference methods) in most patients treated with therapeutic doses of oxcarbazepine¹,²,³." ¹ Flesch, G. 2004. Overview of the clinical pharmacokinetics of oxcarbazepine. Clin Drug Invest 24:185-203. ² Patsalos, P. N. et al. 2008. Antiepileptic drugs – best practice guidelines for therapeutic drug monitoring: A position paper by the subcommission on therapeutic drug monitoring, ILAE Commission on Therapeutic Strategies. Epilepsia 49:1239-1276. ³ Borusiak, P. et al. 1998. Oxcarbazepine in treatment of childhood epilepsy: A survey of 46 children and adolescents. J Epilepsy 11:355-360. 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