Immunalysis Ethyl Alcohol Enzyme Assay

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: k181553 B. Purpose for Submission: New Device C. Measurand: Ethyl Alcohol (Ethanol) D. Type of Test: Automated quantitative enzymatic assay E. Applicant: Immunalysis Corporation F. Proprietary and Established Names: Immunalysis Ethyl Alcohol Enzyme Assay G. Regulatory Information: | Product Code | Classification | Regulation Section | Panel | | --- | --- | --- | --- | | DIC | Class II | 21 CFR 862.3040, Alcohol Test System | Toxicology (91) | H. Intended Use: 1. Intended use(s): Refer to Indications for Use below. 2. Indication(s) for use: The Immunalysis Ethyl Alcohol Enzyme Assay is an in vitro diagnostic device for the quantitative analysis of ethyl alcohol (ethanol) in human urine, serum or plasma with automated clinical chemistry analyzers. The measurement of ethanol is used for the {1} diagnosis and treatment of alcohol intoxication and poisoning. This assay is calibrated against ethyl alcohol. This device is intended for prescription use only. 3. Special conditions for use statement(s): For prescription use only. 4. Special instrument requirements: Beckman Coulter AU400e Chemistry Analyzer I. Device Description: The Immunalysis Ethyl Alcohol Enzyme Assay consists of ready to use reagents on automated clinical chemistry analyzers. The vials of liquid reagents are provided in multiple kit sizes by volume: 25 mL, 60 mL, 100 mL and 500 mL. Each kit box contains the following reagents: - 1 X Reagent 1 (R1) – This contains Tris buffer with 0.1% Sodium Azide as a preservative. - 1 X Reagent 2 (R2) – This contains alcohol dehydrogenase (ADH) and nicotinamide adenine dinucleotide (NAD) in Tris buffer with stabilizers and 0.1% Sodium Azide as a preservative. J. Substantial Equivalence Information: 1. Predicate device name(s): Lin-Zhi Ethyl Alcohol Enzymatic Assay 2. Predicate 510(k) number(s): k032461 3. Comparison with predicate: | Similarities | | | | --- | --- | --- | | Item | Candidate Device: Immunalysis Ethyl Alcohol Enzyme Assay | Predicate Device: Lin-Zhi Ethyl Alcohol Enzymatic Assay (k032461) | | Intended Use | For quantitative analysis of ethyl alcohol in human urine, serum, or plasma. | Same | | Test Principle or Method | Enzymatic assay | Same | {2} | Similarities | | | | --- | --- | --- | | Item | Candidate Device: Immunalysis Ethyl Alcohol Enzyme Assay | Predicate Device: Lin-Zhi Ethyl Alcohol Enzymatic Assay (k032461) | | Sample Type | Urine, serum, or plasma | Same | | Intended Use Environment | For prescription use only | Same | | Assay Measuring Range | 3-550 mg/dL | Same | | Reagent Storage | 2-8°C until expiration date | Same | | Differences | | | | Item | Candidate Device: Immunalysis Ethyl Alcohol Enzyme Assay | Predicate Device: Lin-Zhi Ethyl Alcohol Enzymatic Assay (k032461) | | Analyzer used to collect performance data | Beckman Coulter AU400e analyzer | Hitachi 717 analyzer | # K. Standard/Guidance Document Referenced (if applicable): CLSI EP5-A3: "Evaluation of Precision of Quantitative Measurement Procedures: Approved Guideline-Third Edition" CLSI EP12-A2: "User Protocol for Evaluation of Qualitative Test Performance; Approved Guideline-Second Edition" CLSI EP17-A2: "Evaluation of Detection Capability for Clinical Laboratory Measurement" # L. Test Principle: The Immunalysis Ethanol Alcohol Enzyme Assay technology is based on the oxidation of ethyl alcohol to acetaldehyde by alcohol dehydrogenase (ADH) and nicotinamide adenine dinucleotide (NAD) reduced to NADH resulting in an absorbance change measured spectrophotometrically at $340\mathrm{nm}$ . The concentration of ethanol in the sample is directly proportional to the ADH activity. # M. Performance Characteristics: # 1. Analytical performance: # a. Precision/Reproducibility: The precision studies were performed on drug free urine, serum and plasma spiked with ethanol to concentrations of $25\mathrm{mg / dL}$ , $75\mathrm{mg / dL}$ , $125\mathrm{mg / dL}$ , $150\mathrm{mg / dL}$ , 175 {3} mg/dL and 200 mg/dL, and calibrators and controls at concentrations of 50 mg/dL, 100 mg/dL and 300 mg/dL. The spiked concentrations were confirmed by gas chromatography. Each sample was run in duplicate twice a day for twenty days for a total of 80 measurements at each concentration. The data are summarized in the following tables: Precision - Urine | Conc. (mg/dL) | n | Mean Conc. (mg/dL) | Repeatability | | Between run | | Within -Lab | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | %CV | SD | %CV | SD | %CV | | 0 | 80 | 0.0 | 0.1 | N/A | 0.0 | N/A | 0.1 | N/A | | 25 | 80 | 24.5 | 0.3 | 1.2 | 0.3 | 1.1 | 0.5 | 1.8 | | 50 | 80 | 49.0 | 0.4 | 0.9 | 0.6 | 1.2 | 0.8 | 1.5 | | 75 | 80 | 74.0 | 0.7 | 0.9 | 1.0 | 1.3 | 1.1 | 1.5 | | 100 | 80 | 99.6 | 0.8 | 0.8 | 1.6 | 1.6 | 1.6 | 1.6 | | 125 | 80 | 121.3 | 1.2 | 1.0 | 1.5 | 1.2 | 1.8 | 1.5 | | 150 | 80 | 146.1 | 1.3 | 0.9 | 1.9 | 1.3 | 2.4 | 1.7 | | 175 | 80 | 169.1 | 1.4 | 0.9 | 2.0 | 1.1 | 2.7 | 1.6 | | 200 | 80 | 193.9 | 1.8 | 0.9 | 2.6 | 1.3 | 3.4 | 1.7 | | 300 | 80 | 292.0 | 2.2 | 0.8 | 3.8 | 1.3 | 4.6 | 1.6 | Precision - Serum | Conc. (mg/dL) | n | Mean Conc. (mg/dL) | Repeatability | | Between run | | Within -Lab | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | %CV | SD | %CV | SD | %CV | | 0 | 80 | 0.4 | 0.2 | N/A | 0.1 | N/A | 0.2 | N/A | | 25 | 80 | 25.8 | 0.2 | 0.9 | 0.5 | 2.0 | 0.8 | 3.2 | | 50 | 80 | 49.3 | 0.3 | 0.7 | 1.0 | 2.0 | 1.0 | 2.0 | | 75 | 80 | 75.7 | 0.4 | 0.5 | 1.4 | 1.8 | 2.2 | 3.0 | | 100 | 80 | 99.6 | 0.5 | 0.5 | 2.0 | 2.1 | 2.1 | 2.1 | | 125 | 80 | 125.9 | 0.8 | 0.6 | 2.4 | 1.9 | 3.9 | 3.1 | | 150 | 80 | 148.4 | 1.0 | 0.6 | 2.6 | 1.7 | 3.8 | 2.6 | | 175 | 80 | 172.3 | 1.0 | 0.6 | 3.1 | 1.8 | 5.1 | 3.0 | | 200 | 80 | 195.2 | 1.3 | 0.7 | 3.4 | 1.8 | 4.9 | 2.5 | | 300 | 80 | 290.4 | 1.4 | 0.5 | 5.5 | 1.9 | 6.0 | 2.1 | Precision - Plasma | Conc. (mg/dL) | n | Mean Conc. (mg/dL) | Repeatability | | Between run | | Within -Lab | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | %CV | SD | %CV | SD | %CV | | 0 | 80 | 0.2 | 0.1 | N/A | 0.1 | N/A | 0.1 | N/A | | 25 | 80 | 24.3 | 0.2 | 0.7 | 0.2 | 1.0 | 1.2 | 4.9 | | 50 | 80 | 49.9 | 0.3 | 0.6 | 0.5 | 0.9 | 0.7 | 1.5 | | 75 | 80 | 69.8 | 0.4 | 0.5 | 0.7 | 1.0 | 1.8 | 2.6 | {4} 5 b. Linearity/assay reportable range: To evaluate linearity and recovery, a study was conducted on the Beckman Coulter AU400e analyzer by measuring twelve levels of human urine, serum and plasma samples in triplicate, with the mean of the three measurements used to calculate the recovery. Drug-free human urine, serum and plasma were spiked with ethanol (EtOH) to prepare the high pool at approximately 550 mg/dL, which was confirmed by gas chromatography. The remainder of the samples were prepared by intermixing the high pool with a low pool to obtain concentrations across the measuring range. The linearity results are shown below: Linearity/recovery test – Urine | Expected conc. (mg/dL) | Mean conc. (mg/dL) | %Recovery | | --- | --- | --- | | 0 | 0.0 | n/a | | 3 | 3.1 | 102.2 | | 12 | 12.0 | 100.3 | | 72 | 73.9 | 102.6 | | 132 | 132.2 | 100.2 | | 192 | 195.4 | 101.8 | | 252 | 250.3 | 99.3 | | 312 | 308.0 | 98.7 | | 372 | 363.0 | 97.6 | | 432 | 421.3 | 97.5 | | 492 | 484.4 | 98.5 | | 552 | 565.8 | 102.5 | | Slope | 0.996 | | --- | --- | | Intercept | -0.172 | | r² | 0.999 | Linearity/recovery test – Serum | Expected conc. (mg/dL) | Mean conc. (mg/dL) | %Recovery | | --- | --- | --- | | 0 | 0.0 | n/a | | 3 | 3.0 | 101.1 | | 23.2 | 23.1 | 99.4 | | 83.2 | 87.2 | 104.8 | | 143.2 | 146.1 | 102.0 | {5} | 203.2 | 199.8 | 98.3 | | --- | --- | --- | | 263.2 | 262.2 | 99.6 | | 323.2 | 317.8 | 98.3 | | 383.2 | 385.9 | 100.7 | | 443.2 | 441.4 | 99.6 | | 503.2 | 493.6 | 98.1 | | 563.2 | 544.3 | 96.6 | | Slope | 0.978 | | --- | --- | | Intercept | 2.869 | | r² | 0.999 | ## Linearity/recovery test – Plasma | Expected conc. (mg/dL) | Mean conc. (mg/dL) | %Recovery | | --- | --- | --- | | 0 | 0.0 | n/a | | 3 | 3.0 | 98.9 | | 9.9 | 10.5 | 106.1 | | 69.9 | 69.7 | 99.8 | | 129.9 | 127.5 | 98.2 | | 189.9 | 188.4 | 99.2 | | 249.9 | 242.3 | 96.9 | | 309.9 | 299.9 | 96.8 | | 369.9 | 351.9 | 95.1 | | 429.9 | 410.0 | 95.4 | | 489.9 | 470.4 | 96.0 | | 549.9 | 547.0 | 99.5 | | Slope | 0.970 | | --- | --- | | Intercept | 0.307 | | r² | 0.999 | The results of the linearity testing confirmed the sponsor’s claimed measuring range of $3 - 550\ \mathrm{mg/dL}$ c. Traceability, Stability, Expected values (controls, calibrators, or methods): ### Traceability The controls and calibrators are traceable to the NIST ethanol standard. d. Detection limit: Limit of Blank (LoB): LoB was evaluated by performing sixty (60) blank {6} measurements from unaltered drug free negative human urine, serum and plasma. The Limit of blank (LoB) for each matrix was calculated using two lots, and the higher calculated LoB of the two lots was used as the claimed LoB and to calculate the Limit of Detection (LoD). The claimed Limit of Blank (LoB) of the assay is $0.481\mathrm{mg / dL}$ for urine, $0.668\mathrm{mg / dL}$ for serum and $0.652\mathrm{mg / dL}$ for plasma. Limit of Detection (LoD): Four low level samples of each matrix containing concentrations in the range LoB to $4 \times$ LoB were tested in replicates of five on two reagent lots over three days for a total of 60 replicates per lot. The LoD was calculated according to the formula $\mathrm{LoD} = \mathrm{LoB} + (\mathrm{Cp} \times \mathrm{SD}_{\mathrm{L}})$ where Cp is derived from the 95th percentile of the standard Gaussian distribution and $\mathrm{SD}_{\mathrm{L}}$ is the estimated standard deviation. The sponsor claims an LoD of $1.3 \mathrm{mg} / \mathrm{dL}$ for urine, $1.7 \mathrm{mg} / \mathrm{dL}$ for serum and $1.7 \mathrm{mg} / \mathrm{dL}$ for plasma. Limit of quantitation (LoQ): Four independent samples of each matrix spiked at low concentration of ethanol were measured in two runs in replicates of five for three days on two reagent lots for a total of 60 replicates per lot. The LoQ was calculated based on $\% \mathrm{CV} \leq 10\%$ and Bias $\leq 10\%$ and was determined to be $2.9\mathrm{mg / dL}$ for urine, serum and plasma. # e. Analytical specificity: 1. Interference studies were performed in accordance to CLSI EP07-A2. Test solutions for each compound were prepared by spiking the potential interfering compound into drug-free negative urine containing ethanol concentrations of $10\mathrm{mg / dL}$ and $100\mathrm{mg / dL}$ . The following substances at were tested at the indicated concentrations, and none of these compounds demonstrated significant interference (defined by the sponsor as $\geq 10\%$ bias). Non-Interfering Structurally Unrelated Compounds in Urine: | Compound | Compound Conc. (mg/dL) | | --- | --- | | Acetaminophen | 500,000 | | 6-Acetylcodeine | 100,000 | | 6-Acetylmorphine | 100,000 | | Acetylsalicylic Acid | 500,000 | | Alphenal | 100,000 | | Alprazolam | 100,000 | | 7-Aminoclonazepam | 40,000 | | 7-Aminoflurnitrazepam | 100,000 | | 7-Aminonitrazepam | 100,000 | | Amitriptyline | 100,000 | | Amobarbital | 100,000 | | S-(+)-Amphetamine | 100,000 | | Aprobarbital | 100,000 | | Barbital | 100,000 | {7} | Compound | Compound Conc. (mg/dL) | | --- | --- | | Benzoylecgonine | 100,000 | | Benzylpiperzine | 100,000 | | Bromazepam | 100,000 | | 4-bromo 2-5, dimethoxyphenethylamine | 100,000 | | Buprenorphine | 50,000 | | Bupropion | 100,000 | | Butabarbital | 100,000 | | Butalbital | 100,000 | | Caffeine | 500,000 | | Cannabidiol | 100,000 | | Cannabinol | 100,000 | | Carbamazpine | 100,000 | | Carisoprodol | 100,000 | | Chlordiazepoxide | 100,000 | | Chlorpromazine | 100,000 | | Clobazam | 100,000 | | Clomipramine | 100,000 | | Clonazepam | 100,000 | | Clozapine | 100,000 | | Cocaine | 100,000 | | Codeine | 100,000 | | Cotinine | 100,000 | | Cyclobenzaprine | 100,000 | | Cyclopentobarbital | 100,000 | | Demoxepam | 100,000 | | Deslkylflurazepam | 100,000 | | Desipramine | 100,000 | | Dextromethorphan | 100,000 | | Diazepam | 100,000 | | Digoxin | 100,000 | | Dihydrocodeine | 100,000 | | Diphenhydramine | 500,000 | | Dehydronorketamine | 40,000 | | Delta 9 THC | 100,000 | | Doxepin | 100,000 | | Doxylamine | 100,000 | | ecgonine | 100,000 | | ecgonine methyl ester | 100,000 | | EDDP | 100,000 | | EMDP | 100,000 | | 1R,2S Ephedrine | 100,000 | | 1S,2R Ephendrine | 100,000 | | Ethyl-β-D-Glucuronide | 50,000 | 8 {8} | Compound | Compound Conc. (mg/dL) | | --- | --- | | Ethylmorphine | 100,000 | | Fenfluramine | 100,000 | | Fentanyl | 100,000 | | Flunitrazepam | 100,000 | | Fluoxetine | 100,000 | | Flurazepam | 100,000 | | Haloperidol | 100,000 | | Heroin | 100,000 | | Hexobarbital | 100,000 | | hydrocodone | 100,000 | | hydromorphone | 100,000 | | 11-hydroxy-delta9 THC | 100,000 | | Ibuprofen | 100,000 | | Imipramine | 100,000 | | Ketamine | 100,000 | | Lamotrinine | 100,000 | | Levorphanol Tartrate | 100,000 | | Lidocaine | 100,000 | | Lorazepam | 100,000 | | Lorazepam Glucuronide | 50,000 | | Lormetrazepam | 100,000 | | LSD | 100,000 | | Maprotinline | 100,000 | | MDA | 100,000 | | MDEA | 100,000 | | MDMA | 100,000 | | Meperidine | 100,000 | | Meprobamate | 100,000 | | Methamphetamine | 100,000 | | Methadone | 500,000 | | Methaquolone | 100,000 | | Methoxetamine | 100,000 | | Methylone | 100,000 | | Methylphenidate | 100,000 | | Midazolam | 100,000 | | Morphine | 100,000 | | Morphine-3-glucuronide | 100,000 | | Morphine-6-glucuronide | 100,000 | | N-Desmethyltapentadol | 100,000 | | Nalorphine | 100,000 | | Naloxone | 100,000 | | Naltrexone | 100,000 | | Naproxen | 100,000 | | Nitrazepam | 100,000 | 9 {9} | Compound | Compound Conc. (mg/dL) | | --- | --- | | 11-nor-9-carboxy-delta9-THC | 100,000 | | Norbuprenorphine | 50,000 | | Norcodeine | 100,000 | | Nordiazepam | 100,000 | | Norketamine | 100,000 | | Normorphine | 100,000 | | Norproxyphene | 100,000 | | Norpseudoephedrine | 50,000 | | Nortripyline | 100,000 | | N-desmethyltramadol | 100,000 | | N-desmethylvenlafaxine | 100,000 | | O-desmethyltramadol | 100,000 | | O-desmethylvenlafaxine | 100,000 | | Olanzapine | 100,000 | | Oxycodone | 100,000 | | Oxymorphone | 100,000 | | Oxazepam | 100,000 | | PCP | 100,000 | | Pentazocine | 100,000 | | Pentobarbital | 100,000 | | Phenazepam | 100,000 | | Phenobarbital | 100,000 | | Phentermine | 100,000 | | Phenylephedrine | 100,000 | | Phenylpropanolamine | 100,000 | | Phenytoin | 100,000 | | PMA | 100,000 | | Prazepam | 100,000 | | Propranolol | 100,000 | | Propoxyphene | 100,000 | | Protripyline | 100,000 | | R,R Psuedoephedrine | 100,000 | | S,S Psuedoephedrine | 100,000 | | Ritalinic Acid | 100,000 | | Salicylic Acid | 100,000 | | Sertaraline | 100,000 | | Sufentanil Citrate | 50,000 | | Talbutal | 50,000 | | Tapentadol | 100,000 | | Temazepam | 100,000 | | Theophylline | 100,000 | | Thiopental | 100,000 | | Thiordazine | 100,000 | | Tramadol | 100,000 | {10} | Compound | Compound Conc. (mg/dL) | | --- | --- | | Trazadone | 100,000 | | Triazolam | 100,000 | | Trifluoromethylphenyl-piperazine | 100,000 | | Trimipramine | 100,000 | | Venlafaxine | 100,000 | | Verapamil | 100,000 | | Zolpidem Tartrate | 100,000 | | O-desmethylvenlafaxine | 100,000 | | Olanzapine | 100,000 | | Oxycodone | 100,000 | | Oxymorphone | 100,000 | | Oxazepam | 100,000 | | PCP | 100,000 | | Pentazocine | 100,000 | | Pentobarbital | 100,000 | | Phenazepam | 100,000 | | Phenobarbital | 100,000 | | Phentermine | 100,000 | | Phenylephedrine | 100,000 | | Phenylpropanolamine | 100,000 | | Phenytoin | 100,000 | | PMA | 100,000 | | Prazepam | 100,000 | | Propranolol | 100,000 | | Propoxyphene | 100,000 | | Protripyline | 100,000 | | R,R Psuedoephedrine | 100,000 | | S,S Psuedoephedrine | 100,000 | | Ritalinic Acid | 100,000 | | Salicylic Acid | 100,000 | | Sertaraline | 100,000 | | Sufentanil Citrate | 50,000 | | Talbutal | 50,000 | | Tapentadol | 100,000 | | Temazepam | 100,000 | | Theophylline | 100,000 | | Thiopental | 100,000 | | Thiordazine | 100,000 | | Tramadol | 100,000 | | Trazadone | 100,000 | | Triazolam | 100,000 | | Trifluoromethylphenyl-piperazine | 100,000 | | Trimipramine | 100,000 | | Venlafaxine | 100,000 | {11} | Compound | Compound Conc. (mg/dL) | | --- | --- | | Verapamil | 100,000 | | Zolpidem Tartrate | 100,000 | Non-Interfering Endogenous Compounds and Urine Preservatives | Compound | 10 mg/dL and 100 mg/dL | | --- | --- | | | Compound Conc. | | Ascorbic Acid | 1.5 g/dL | | Bilirubin | 0.02 g/dL | | Creatinine | 0.5 g/dL | | Galactose | 0.01 g/dL | | γ-Globulin | 0.5 g/dL | | Glucose | 2.0 g/dL | | Hemoglobin | 0.3 g/dL | | Human Serum Albumin | 0.5 g/dL | | Oxalic Acid | 0.1 g/dL | | Riboflavin | 0.0075 g/dL | | Sodium Chloride | 6.0 g/dL | | Urea | 6.0 g/dL | | Boric Acid | 1% w/v | | Sodium Azide | 1% w/v | | Sodium Fluoride | 1% w/v | Non-interfering Substances - Serum and Plasma | Compound | 10 mg/dL | 100 mg/dL | | --- | --- | --- | | | Compound Conc. | Compound Conc. | | Acetaminophen | 20 mg/dL | 20 mg/dL | | Amikacin | 8 mg/dL | 8 mg/dL | | Ampicillin | 5.3 mg/dL | 5.3 mg/dL | | Ascorbic Acid | 6 mg/dL | 6 mg/dL | | Bilirubin total | 60 mg/dL | 80 mg/dL | | Bilirubin direct | 60 mg/dL | 80 mg/dL | | Caffeine | 6 mg/dL | 6 mg/dL | | Carbamazepine | 3 mg/dL | 3 mg/dL | | Chloramphenicol | 5 mg/dL | 5 mg/dL | | Chlordiazepoxide | 1 mg/dL | 1 mg/dL | | Chlorpromazine | 0.2 mg/dL | 0.2 mg/dL | | Cholesterol | 30 mg/dL | 503 mg/dL | | Cimetidine | 2 mg/dL | 2 mg/dL | | Creatinine | 30 mg/dL | 30 mg/dL | | Dextran | 6000 mg/dL | 6000 mg/dL | | Diazepam | 0.51 mg/dL | 0.51 mg/dL | {12} | Compound | 10 mg/dL | 100 mg/dL | | --- | --- | --- | | | Compound Conc. | Compound Conc. | | Digoxin | 6.1 mg/dL | 6.1 mg/dL | | Erythromycin | 6 mg/dL | 6 mg/dL | | Ethosuximide | 25 mg/dL | 25 mg/dL | | Furosemide | 6 mg/dL | 6 mg/dL | | Gentamicin | 1 mg/dL | 1 mg/dL | | Hemoglobin | 62.5 mg/dL | 1,000 mg/dL | | Heparin | 3 U/mL | 3 U/mL | | Ibuprofen | 50 mg/dL | 50 mg/dL | | IgG | 5,000 mg/dL | 5,000 mg/dL | | Intralipid | 187.5 mg/dL | 750 mg/dL | | Lactate Dehydrogenase | 1,000 U/L | 237,500 U/L | | Lactate | 150 mg/dL | 901 mg/dL | | Lidocaine | 1.2 mg/dL | 1.2 mg/dL | | Lithium | 2.2 mg/dL | 2.2 mg/dL | | Mannitol | 500 mg/dL | 500 mg/dL | | Penicillin | 25 U/mL | 25 U/mL | | Pentobarbital | 8 mg/dL | 8 mg/dL | | Phenobarbital | 10 mg/dL | 10 mg/dL | | Phenytoin | 5 mg/dL | 5 mg/dL | | Primidone | 4 mg/dL | 4 mg/dL | | Propoxyphene | 0.16 mg/dL | 0.16 mg/dL | | HSA (albumin) | 37.5 mg/dL | 1,250 mg/dL | | Protein (total) | 50 mg/dL | 1,500 mg/dL | | Salicylic Acid | 60 mg/dL | 60 mg/dL | | Theophylline | 4 mg/dL | 4 mg/dL | | Triglycerides | 500 mg/dL | 500 mg/dL | | Urea | 500 mg/dL | 500 mg/dL | | Uric Acid | 20 mg/dL | 20 mg/dL | | Valproic Acid | 50 mg/dL | 50 mg/dL | Non-interfering Anticoagulants -Plasma | Compound | 10 mg/dL | 100 mg/dL | | --- | --- | --- | | | Compound Conc. | Compound Conc. | | Heparin | 3 U/mL | 3 U/mL | | EDTA | 2 mg/mL | 2 mg/mL | | Oxalic Acid | 2 mg/mL | 2 mg/mL | | Sodium Citrate | 3.8 g/dL | 3.8 g/dL | | Sodium Fluoride | 2 mg/mL | 2 mg/mL | 2. To evaluate cross-reactivity with structurally similar compounds, the potential cross-reactants were spiked into alcohol free urine, serum, and plasma and measured for ethanol using the candidate assay. Results were as follows: {13} | Compound | Matrix | % cross-reactivity | | --- | --- | --- | | Acetaldehyde | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | | Acetone | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | | n-Butanol | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | | Ethylene Glycol | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | | Isopropanol | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | | Methanol | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | | n-Propanol | Urine | 16.7 | | | Serum | 11.1 | | | Plasma | 11.1 | | Propylene Glycol | Urine | <3.3 | | | Serum | <3.3 | | | Plasma | <3.3 | # 3. Interference - pH To evaluate potential interference from the effect of urine pH, device performance in the quantitative mode was tested using a range of urine pH values. All test samples were prepared in drug-free urine containing EtOH at concentration of $10\mathrm{mg / dL}$ and $100\mathrm{mg / dL}$ . No interference was observed at urine pH values ranging from 3.0 to 11.0. {14} 4. Interference - Specific Gravity To evaluate potential interference from the specific gravity of urine, device performance in the quantitative mode was tested using a range of physiologically relevant urine specific gravity values. All test samples were prepared in drug free urine containing EtOH at concentration of $10\mathrm{mg / dL}$ and $100\mathrm{mg / dL}$ . No interference was observed at urine specific gravity values ranging from 1.000 to 1.030. f. Assay cut-off: Not applicable. # 2. Comparison studies: a. Method comparison with predicate device: A method comparison study was performed using unaltered clinical urine, serum and plasma samples. 80 urine samples, 76 serum samples and 64 plasma samples with concentrations covering the measuring range of candidate device were evaluated. Results were compared to the predicate device, the Lin-Zhi Ethyl Alcohol Enzymatic Assay, using linear regression and the study results are summarized in the table below. | Matrix | n | Slope | Intercept | r2 | | --- | --- | --- | --- | --- | | Urine | 80 | 1.0284 | -2.7848 | 0.9964 | | Serum | 76 | 1.0067 | -5.8802 | 0.9948 | | Plasma | 64 | 1.0595 | -7.3902 | 0.9932 | 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. {15} 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. 16