LZI Buprenorphine II Enzyme Immunoassay

{0} FDA U.S. FOOD &amp; DRUG ADMINISTRATION # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY ## I Background Information: A 510(k) Number K253082 B Applicant Lin-Zhi International, Inc C Proprietary and Established Names Proprietary name: LZI Buprenorphine II Enzyme Immunoassay Common name: Homogeneous Buprenorphine Enzyme Immunoassay D Regulatory Information | Product Code(s) | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | DJG | II | 21 CFR 862.3650 | 91-Toxicology | ## II Submission/Device Overview: A Purpose for Submission: Modification to previously cleared assay B Measurand: Norbuprenorphine (buprenorphine metabolite) C Type of Test: Homogeneous enzyme immunoassay – qualitative and semi-quantitative ## III Intended Use/Indications for Use: A Intended Use(s): See Indications for Use below. B Indication(s) for Use: The LZI Buprenorphine II Enzyme Immunoassay is intended for the qualitative and semi-quantitative determination of norbuprenorphine in human urine at the cutoff Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993-0002 www.fda.gov {1} value of 5 ng/mL when calibrated against norbuprenorphine. The assay is designed for prescription 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 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 chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. Clinical consideration and professional judgment should be exercised with any drug of abuse test result, particularly when the preliminary test result is positive. ## C Special Conditions for Use Statement(s): The assay is for prescription use. ## D Special Instrument Requirements: Automated clinical chemistry analyzers. (Performance in the studies submitted in the 510(k) are based on the Beckman Coulter AU480 Analyzer. ## IV Device/System Characteristics: ### A Device Description: The LZI Buprenorphine II Enzyme Immunoassay is a kit comprised of two reagents, R1 and R2, which are bottled separately but sold together within the kit. The R1 solution contains mouse monoclonal anti-norbuprenorphine 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 norbuprenorphine in buffer with sodium azide (0.09%) as a preservative. ### B Principle of Operation: The LZI Buprenorphine II Enzyme Immunoassay is a homogeneous enzyme immunoassay ready-to-use liquid reagent. The assay is based on competition between drug in the sample and drug labeled with the 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, norbuprenorphine-labeled G6PDH conjugate is bound to antibody, and the enzyme activity is inhibited. On the other hand, when drug is present in the sample, antibody would bind to free drug; the unbound norbuprenorphine-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. K253082 - Page 2 of 14 {2} V Substantial Equivalence Information: A Predicate Device Name(s): LZI Buprenorphine Enzyme Immunoassay B Predicate 510(k) Number(s): k090844 C Comparison with Predicate(s): | Device & Predicate Device(s): | K253082 | K090844 | | --- | --- | --- | | Device Trade Name | LZI Buprenorphine II Enzyme Immunoassay | LZI Buprenorphine Enzyme Immunoassay | | General Device Characteristic Similarities | | | | Intended Use | For the qualitative and semi-quantitative determination of norbuprenorphine in human urine at the cutoff value of 5 ng/mL when calibrated against norbuprenorphine. The assay is designed for prescription use with a number of automated clinical chemistry analyzers. The assay provides only a preliminary analytical result. A more specific alternative chemical method (e.g., gas or liquid chromatography and mass spectrometry) must be used in order to obtain a confirmed analytical result. | Same | K253082 - Page 3 of 14 {3} | Analyte | norbuprenorphine | Same | | --- | --- | --- | | Matrix | Urine | Same | | Storage | 2-8 °C until expiration date | Same | | Detection | Absorbance change measured spectrophotometrically at 340 nm | Same | | User Environment | Clinical laboratories: Prescription use only | Same | | Mass Spectrometry Confirmation | Required to confirm preliminary positive analytical results | Same | | Platform Required | Automated clinical chemistry analyzer | Same | | Reagents Form | Liquid – ready-to-use | Same | | General Device Characteristic Differences | | | | Cutoff | 5 ng/mL | 5 ng/mL and 10 ng/mL | | Instrument for performance validation | Beckman Coulter AU480 | Hitachi 717 | | Calibrator Level | 0, 2.5, 5, 10, and 20 ng/mL | 0, 5, 10, 20, 40, and 75 ng/mL | | Controls Level | 5 ng/mL Cutoff: 2 Levels: 3.75 and 6.25 ng/mL 10 ng/mL Cutoff: 2 Levels 7 and 13 ng/mL | 5 ng/mL Cutoff: 2 Levels: 3 and 7 ng/mL 10 ng/mL Cutoff: 2 Levels 7 and 13 ng/mL | VI Standards/Guidance Documents Referenced: No device-specific guidance documents or special controls are applicable to the LZI Buprenorphine II Enzyme Immunoassay under the classification regulation 21 CFR 862.3650 (Product Code: DJG). VII Performance Characteristics (if/when applicable): K253082 - Page 4 of 14 {4} All 510(k) studies below were conducted on the Beckman Coulter AU480 Analyzer A Analytical Performance: 1. Precision/Reproducibility: Spiked samples of norbuprenorphine were prepared in a urine matrix at the concentrations shown in the table below, and were measured, 2 runs per day, with 2 replicates per run, for 22 days. Results are tabulated below: Precision: 5 ng/mL Cutoff Semi-Quantitative Positive/Negative Results: | 5 ng/mL Cutoff Result: | | Within Run (N=22) | | Total Precision (N=88) | | | --- | --- | --- | --- | --- | --- | | Norbuprenorphine Concentration | % of Cutoff | Number of Determination | Immunoassay Result | Number of Determination | Immunoassay Result | | 0 ng/mL | 0% | 22 | 22 Negative | 88 | 88 Negative | | 1.25 ng/mL | 25% | 22 | 22 Negative | 88 | 88 Negative | | 2.5 ng/mL | 50% | 22 | 22 Negative | 88 | 88 Negative | | 3.75 ng/mL | 75% | 22 | 22 Negative | 88 | 88 Negative | | 5 ng/mL | 100% | 22 | 22 Positive | 88 | 2 Neg / 86 Pos | | 6.25 ng/mL | 125% | 22 | 22 Positive | 88 | 88 Positive | | 7.5 ng/mL | 150% | 22 | 22 Positive | 88 | 88 Positive | | 8.75 ng/mL | 175% | 22 | 22 Positive | 88 | 88 Positive | | 10 ng/mL | 200% | 22 | 22 Positive | 88 | 88 Positive | Semi-Quantitative Precision Analysis Summary (ng/mL): | Norbuprenorphine concentration | Within Run (N=22) | | Total Precision (N=88) | | | --- | --- | --- | --- | --- | | | SD | %CV | SD | %CV | | 0 ng/mL | 0.15 | N/A | 0.2 | N/A | | 1.25 ng/mL | 0.17 | 12.0% | 0.21 | 14.6% | | 2.5 ng/mL | 0.17 | 6.4% | 0.24 | 8.8% | | 3.75 ng/mL | 0.19 | 4.7% | 0.28 | 6.9% | | 5 ng/mL | 0.19 | 3.5% | 0.24 | 4.4% | | 6.25 ng/mL | 0.19 | 2.8% | 0.27 | 4.0% | | 7.5 ng/mL | 0.2 | 2.5% | 0.24 | 3.0% | | 8.75 ng/mL | 0.21 | 2.2% | 0.26 | 2.8% | | 10 ng/mL | 0.3 | 2.8% | 0.36 | 3.4% | K253082 - Page 5 of 14 {5} Qualitative Positive/Negative Results: | 5 ng/mL Cutoff Result: | | 5 ng/mL Cutoff Result: | | Total Precision (N=88) | | | --- | --- | --- | --- | --- | --- | | Norbuprenorphine Concentration | % of Cutoff | Number of Determination | Immunoassay Result | Number of Determination | Immunoassay Result | | 0 ng/mL | 0% | 22 | 22 Negative | 88 | 88 Negative | | 1.25 ng/mL | 25% | 22 | 22 Negative | 88 | 88 Negative | | 2.5 ng/mL | 50% | 22 | 22 Negative | 88 | 88 Negative | | 3.75 ng/mL | 75% | 22 | 22 Negative | 88 | 88 Negative | | 5 ng/mL | 100% | 22 | 1 Neg / 21 Pos | 88 | 19 Neg / 69 Pos | | 6.25 ng/mL | 125% | 22 | 22 Positive | 88 | 88 Positive | | 7.5 ng/mL | 150% | 22 | 22 Positive | 88 | 88 Positive | | 8.75 ng/mL | 175% | 22 | 22 Positive | 88 | 88 Positive | | 10 ng/mL | 200% | 22 | 22 Positive | 88 | 88 Positive | 2. Linearity/Reportable range: To demonstrate linearity in the semiquantitative mode, which is used for purposes of sample dilution and quality control procedures, a drug-free urine pool spiked with norbuprenorphine 20 ng/mL was serially diluted in increments of 10% and results were obtained. The recovery of norbuprenorphine spiked to various concentrations was evaluated and the average recovery from these 10 replicates was used to determine the functional linearity range of the assay. The average recovery across the linear range of the assay was found to range between 99.6% - 106.3%. 3. Analytical Specificity/Interference: Various potentially interfering substances were evaluated for positive and negative interference with the assay. Test compounds were spiked into a buprenorphine-free sample, as well as samples containing buprenorphine at 3.75 ng/mL or 6.25 ng/mL, (as negative or positive controls, ±25% of the cutoff concentration, respectively). Samples were then evaluated against the assay's calibration curve in qualitative mode. Negative interference was observed with Boric Acid at 1% w/v. at ±50% of the cutoff concentration. No other significant cross-reactivity was observed. The potential interferents and the concentrations at which they were tested are shown below: | Interfering Substance | Concentration of Compound (mg/dL) | | --- | --- | | Acetone | 1,000 | | Ascorbic acid | 500 | | Bilirubin | 2 | | Biotin | 2 | | Boric acid | 1,000 | | Calcium chloride | 300 | | Citric acid | 200 | | Creatinine | 500 | K253082 - Page 6 of 14 {6} K253082 - Page 7 of 14 | Ethanol | 1,000 | | --- | --- | | Galactose | 10 | | γ-Globulin | 500 | | Glucose | 3,000 | | Hemoglobin | 300 | | Human urine (pooled) | N/A | | Human serum albumin | 500 | | β-Hydroxybutyric acid | 100 | | Oxalic acid | 100 | | Potassium chloride | 1,000 | | Riboflavin | 7.5 | | Sodium azide | 1,000 | | Sodium chloride | 1,000 | | Sodium fluoride | 1,000 | | Sodium phosphate | 300 | | Urea | 6,000 | | Uric acid | 10 | | Urine-based calibrator buffer | N/A | Cross reactivity of various potentially interfering drugs was evaluated by spiking various concentrations of each substance up to 100,000 ng/mL of buprenorphine metabolites into a pool of drug-free human urine. Results for both 5 and 10 cutoffs are shown below: ## Cross reactivity: 5 cutoff ### Buprenorphine and Major Metabolites: | Compound | Test Concentration (ng/mL) | Semi-Quantitative Result (ng/mL) | % Cross-reactivity | | --- | --- | --- | --- | | Norbuprenorphine | 10 | 10.5 | 100.0% | | Buprenorphine | 10 | 10.5 | 100.0% | | Buprenorphine Glucuronide | 105 | 12.0 | 9.5% | | Norbuprenorphine Glucuronide | 600 | 10.5 | 1.7% | The sponsor also tested a list of structurally related and un-related Opiate compounds. Results are shown below: ### Structurally related or unrelated Opiate compounds: | Compound | Test Concentration (ng/mL) | Qualitative Result (mAU) | Semi-Quantitative Result (ng/mL) | % Cross-reactivity | | --- | --- | --- | --- | --- | {7} K253082 - Page 8 of 14 | Codeine | 100,000 | -17.2 | -0.98 | ND | | --- | --- | --- | --- | --- | | Codeine 6β D-Glucuronide | 100,000 | 0.0 | 0.18 | ND | | Dextromethorphan | 100,000 | 14.3 | 1.43 | ND | | Dextrorphan Tartrate | 100,000 | -7.3 | -0.15 | ND | | Dihydrocodeine | 100,000 | 3.5 | 0.51 | ND | | Dihydromorphine | 100,000 | -7.4 | -0.23 | ND | | EDDP | 100,000 | -6.0 | -0.08 | ND | | EMDP | 100,000 | 12.1 | 1.28 | ND | | Ethylmorphine | 100,000 | -8.0 | -0.15 | ND | | Fentanyl | 100,000 | -22.4 | -0.66 | ND | | Heroin | 100,000 | -8.8 | -0.09 | ND | | Hydrocodone | 100,000 | -18.4 | -0.63 | ND | | Hydromorphone | 100,000 | -20.8 | -0.81 | ND | | Hydromorphone 3β D- Glucuronide | 10,000 | -5.0 | -0.02 | ND | | LAAM | 100,000 | 12.3 | 1.33 | ND | | Levorphanol | 100,000 | -5.8 | -0.05 | ND | | Meperidine | 100,000 | 12.6 | 1.38 | ND | | Meperidine | 100,000 | -15.6 | -0.68 | ND | | Methadone | 100,000 | -18.7 | -0.73 | ND | | Morphine | 100,000 | -11.8 | -0.73 | ND | | Morphine 3β D-Glucuronide | 100,000 | -6.0 | -0.32 | ND | | Morphine 6 D-Glucuronide | 100,000 | -3.1 | 0.24 | ND | | N-Desmethyl-cis-Tramadol | 100,000 | -7.5 | -0.10 | ND | | Nalbuphine | 100,000 | -5.8 | -0.21 | ND | | Nalorphine | 100,000 | -2.1 | 0.12 | ND | | Naloxegol | 100,000 | 36.9 | 1.86 | ND | | Naloxone | 100,000 | 1.0 | -0.20 | ND | | Naltrexone | 100,000 | 15.2 | 1.26 | ND | | Norcodeine | 100,000 | -8.9 | -0.35 | ND | | Norhydrocodone | 100,000 | -7.4 | -0.09 | ND | | Normorphine | 100,000 | -8.6 | -0.27 | ND | | Noroxycodone | 100,000 | -4.6 | -0.05 | ND | | Noroxymorphone | 100,000 | -4.5 | 0.29 | ND | | Norpropoxyphene | 100,000 | 12.0 | 1.40 | ND | | O-Desmethyl-cis-Tramadol | 100,000 | -8.5 | -0.36 | ND | | Oxycodone | 100,000 | -13.5 | -0.57 | ND | | Oxymorphone | 100,000 | -15.6 | -0.80 | ND | | Oxymorphone 3β D- Glucuronide | 10,000 | -3.9 | 0.14 | ND | | Pentazocine | 100,000 | -8.2 | -0.32 | ND | {8} Structurally Unrelated Pharmacological Compounds: | Compound | Test Concentration (ng/mL) | 0 ng/mL Norbuprenorphine | -25% Norbuprenorphine Cutoff (3.75 ng/mL) | +25% Norbuprenorphine Cutoff (6.25 ng/mL) | | --- | --- | --- | --- | --- | | | | % Cross | Result | Result | | Acetaminophen | 100,000 | Neg | Neg | Pos | | 6-Acetylmorphine | 100,000 | Neg | Neg | Pos | | Acetylsalicylic Acid | 100,000 | Neg | Neg | Pos | | Amitriptyline | 100,000 | Neg | Neg | Pos | | Amlodipine Besylate | 100,000 | Neg | Neg | Pos | | Amoxicillin | 100,000 | Neg | Neg | Pos | | d-Amphetamine | 100,000 | Neg | Neg | Pos | | Atorvastatin | 100,000 | Neg | Neg | Pos | | Benzoylecgonine | 100,000 | Neg | Neg | Pos | | Bupropion | 100,000 | Neg | Neg | Pos | | Caffeine | 100,000 | Neg | Neg | Pos | | Carbamazepine | 100,000 | Neg | Neg | Pos | | Cetirizine | 100,000 | Neg | Neg | Pos | | Chlorpheniramine | 100,000 | Neg | Neg | Pos | | Chlorpromazine | 100,000 | Neg | Neg | Pos | | Clomipramine | 100,000 | Neg | Neg | Pos | | Desipramine | 100,000 | Neg | Neg | Pos | | Diphenhydramine | 100,000 | Neg | Neg | Pos | | Duloxetine | 100,000 | Neg | Neg | Pos | | Fluoxetine | 100,000 | Neg | Neg | Pos | | Fluphenazine | 100,000 | Neg | Neg | Pos | | Gabapentin | 100,000 | Neg | Neg | Pos | | Ibuprofen | 100,000 | Neg | Neg | Pos | | Imipramine | 100,000 | Neg | Neg | Pos | | Lisinopril | 100,000 | Neg | Neg | Pos | | Losartan | 100,000 | Neg | Neg | Pos | | Loratidine | 100,000 | Neg | Neg | Pos | | MDA (3,4-methylenedioxyamphetamine) | 100,000 | Neg | Neg | Pos | | MDEA | 100,000 | Neg | Neg | Pos | | MDMA (3,4-methylenedioxymethamphetamine) | 100,000 | Neg | Neg | Pos | | Metformin | 100,000 | Neg | Neg | Pos | K253082 - Page 9 of 14 {9} Structurally related or unrelated Opiate compounds: | Compound | Test Concentration (ng/mL) | Qualitative Result (mAU) | Semi-Quantitative Result (ng/mL) | % Cross-reactivity | | --- | --- | --- | --- | --- | | Codeine | 100,000 | -7.6 | -1.2 | ND | | Codeine 6β D-Glucuronide | 100,000 | -2.0 | 0.1 | ND | | Dextromethorphan | 100,000 | 5.6 | 1.4 | ND | | Dextrorphan Tartrate | 100,000 | -2.9 | -0.7 | ND | | Dihydrocodeine | 100,000 | 2.0 | 0.6 | ND | | Dihydromorphine | 100,000 | 0.2 | -0.1 | ND | | EDDP | 100,000 | -4.2 | -0.4 | ND | | EMDP | 100,000 | 5.1 | 1.0 | ND | | Ethylmorphine | 100,000 | -4.9 | -0.4 | ND | | Fentanyl | 100,000 | -8.9 | -1.7 | ND | | Heroin | 100,000 | -4.6 | -0.7 | ND | | Hydrocodone | 100,000 | -5.7 | -1.0 | ND | | Hydromorphone | 100,000 | -7.6 | -1.3 | ND | | Hydromorphone 3β D-Glucuronide | 10,000 | -2.2 | -0.2 | ND | | LAAM | 100,000 | 5.9 | 1.4 | ND | | Levorphanol | 100,000 | -3.2 | -0.4 | ND | | Meperidine | 100,000 | 4.6 | 0.9 | ND | | Meperidine | 100,000 | -16.3 | -1.1 | ND | | Methadone | 100,000 | -16.4 | -1.3 | ND | | Morphine | 100,000 | -14.2 | -1.0 | ND | | Morphine 3β D-Glucuronide | 100,000 | -4.8 | -0.1 | ND | | Metoprolol | 100,000 | Neg | Neg | Pos | | --- | --- | --- | --- | --- | | d-Methamphetamine | 100,000 | Neg | Neg | Pos | | Nalmefene | 100,000 | Neg | Neg | Pos | | Nicotine | 100,000 | Neg | Neg | Pos | | Nortriptyline | 100,000 | Neg | Neg | Pos | | Omeprazole | 100,000 | Neg | Neg | Pos | | Oxazepam | 100,000 | Neg | Neg | Pos | | Phenobarbital | 100,000 | Neg | Neg | Pos | | (1S,2S)-(+)Pseudoephedrine | 100,000 | Neg | Neg | Pos | | Quetiapine | 100,000 | Neg | Neg | Pos | | Ranitidine | 100,000 | Neg | Neg | Pos | | Salbutamol (Albuterol) | 100,000 | Neg | Neg | Pos | | Sertraline | 100,000 | Neg | Neg | Pos | | THC-COOH (11-Nor-Delta-9-THC-9-carboxylic acid) | 100,000 | Neg | Neg | Pos | | L-Thyroxine | 100,000 | Neg | Neg | Pos | | Zolpidem | 100,000 | Neg | Neg | Pos | K253082 - Page 10 of 14 {10} Structurally Unrelated Pharmacological Compounds: | Compound | Test Concentration (ng/mL) | 0 ng/mL Norbuprenorphine | -25% Norbuprenorphine Cutoff (3.75 ng/mL) | +25% Norbuprenorphine Cutoff 6.25 ng/mL) | | --- | --- | --- | --- | --- | | | | % Cross | Result | Result | | Acetaminophen | 100,000 | Neg | Neg | Pos | | 6-Acetylmorphine | 100,000 | Neg | Neg | Pos | | Acetylsalicylic Acid | 100,000 | Neg | Neg | Pos | | Amitriptyline | 100,000 | Neg | Neg | Pos | | Amlodipine Besylate | 100,000 | Neg | Neg | Pos | | Amoxicillin | 100,000 | Neg | Neg | Pos | | d-Amphetamine | 100,000 | Neg | Neg | Pos | | Atorvastatin | 100,000 | Neg | Neg | Pos | | Benzoylecgonine | 100,000 | Neg | Neg | Pos | | Bupropion | 100,000 | Neg | Neg | Pos | | Caffeine | 100,000 | Neg | Neg | Pos | | Carbamazepine | 100,000 | Neg | Neg | Pos | | Cetirizine | 100,000 | Neg | Neg | Pos | | Chlorpheniramine | 100,000 | Neg | Neg | Pos | | Chlorpromazine | 100,000 | Neg | Neg | Pos | | Clomipramine | 100,000 | Neg | Neg | Pos | K253082 - Page 11 of 14 {11} | Desipramine | 100,000 | Neg | Neg | Pos | | --- | --- | --- | --- | --- | | Diphenhydramine | 100,000 | Neg | Neg | Pos | | Duloxetine | 100,000 | Neg | Neg | Pos | | Fluoxetine | 100,000 | Neg | Neg | Pos | | Fluphenazine | 100,000 | Neg | Neg | Pos | | Gabapentin | 100,000 | Neg | Neg | Pos | | Ibuprofen | 100,000 | Neg | Neg | Pos | | Imipramine | 100,000 | Neg | Neg | Pos | | Lisinopril | 100,000 | Neg | Neg | Pos | | Losartan | 100,000 | Neg | Neg | Pos | | Loratidine | 100,000 | Neg | Neg | Pos | | MDA (3,4-methylenedioxyamphetamine) | 100,000 | Neg | Neg | Pos | | MDEA | 100,000 | Neg | Neg | Pos | | MDMA (3,4-methylenedioxymethamphetamine) | 100,000 | Neg | Neg | Pos | | Metformin | 100,000 | Neg | Neg | Pos | | Metoprolol | 100,000 | Neg | Neg | Pos | | d-Methamphetamine | 100,000 | Neg | Neg | Pos | | Nalmefene | 100,000 | Neg | Neg | Pos | | Nicotine | 100,000 | Neg | Neg | Pos | | Nortriptyline | 100,000 | Neg | Neg | Pos | | Omeprazole | 100,000 | Neg | Neg | Pos | | Oxazepam | 100,000 | Neg | Neg | Pos | | Phenobarbital | 100,000 | Neg | Neg | Pos | | (1S,2S)-(+)Pseudoephedrine | 100,000 | Neg | Neg | Pos | | Quetiapine | 100,000 | Neg | Neg | Pos | | Ranitidine | 100,000 | Neg | Neg | Pos | | Salbutamol (Albuterol) | 100,000 | Neg | Neg | Pos | | Sertraline | 100,000 | Neg | Neg | Pos | | THC-COOH (11-Nor-Delta-9-THC-9-carboxylic acid) | 100,000 | Neg | Neg | Pos | | L-Thyroxine | 100,000 | Neg | Neg | Pos | | Zolpidem | 100,000 | Neg | Neg | Pos | Among all structurally unrelated pharmacological compounds tested, no cross-reactivity was observed with any compound tested at -25% of the norbuprenorphine cutoff concentration with the 5 ng/mL cutoff. Samples ranging in specific gravity from 1.000 to 1.030 were split into three portions each and either left un-spiked or further spiked to a final buprenorphine concentration of either 3.75 ng/mL or 6.25 ng/mL. Samples were evaluated in both qualitative and semi-quantitative modes. No interference was observed. K253082 - Page 12 of 14 {12} Negative urine and urine spiked with norbuprenorphine to the final buprenorphine concentration of either 3.75 ng/mL or 6.25 ng/mL were assessed for interference from pH. No interference was observed between pH of 3 to 11. 4. Detection Limit: The lowest concentration that can be differentiated from the negative urine with 95% confidence is determined as 1.0 ng/mL 5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods): Traceable to a commercially available standard. 6. Assay Cut-Off: See Precision section above. ## B Comparison Studies: 1. Method Comparison with Predicate Device: A total of ninety-four (94) unaltered clinical samples were tested with the LZI Buprenorphine II Enzyme Immunoassay on the Beckman Coulter AU480 automated clinical analyzer. All samples were tested in singlet. All samples were confirmed with LC/MS for both buprenorphine and norbuprenorphine concentrations. Results are shown in the tables below. Semi-Quantitative Accuracy Study: | BUP II Results 5 ng/mL Cutoff | Negative by LC/MS analysis | < 50% of the cutoff concentration by LC/MS analysis | Near Cutoff Negative between 50% below the cutoff and the cutoff concentration by LC/MS analysis | Near Cutoff Positive between the cutoff and 50% above the cutoff concentration by LCMS analysis | High Positive greater than 50% above the cutoff concentration by LC/MS analysis | | --- | --- | --- | --- | --- | --- | | Positive at or above the cutoff by EIA analysis | 0 | 7* | 12* | 14 | 33 | | Negative below the cutoff by EIA analysis | 20 | 6 | 2 | 0 | 0 | * These samples were confirmed to contain known cross reactants (buprenorphine glucuronide and norbuprenorphine glucuronide), which significantly contributed to the discrepant results observed. K253082 - Page 13 of 14 {13} 2. Matrix Comparison: Not applicable. C Clinical Studies: 1. Clinical Sensitivity: Not applicable. 2. Clinical Specificity: Not applicable. 3. Clinical Cut-Off: Not applicable 4. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable): Not applicable. D Expected Values/Reference Range: Not applicable. VIII Proposed Labeling: The labeling supports the finding of substantial equivalence for this device. IX Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision. K253082 - Page 14 of 14