ETHANOL 3L36-20 REAGENT HANDLING AND STORAGE

Transcription

1 ETHANOL 3L36-20 NOTE: This package insert must be read carefully prior to product use. Package insert instructions must be followed accordingly. Reliability of assay results cannot be guaranteed if there are any deviations from the instructions in this package insert. NOTE: Changes Highlighted INTENDED USE The MULTIGENT Ethanol assay is intended for the quantitative determination of ethanol in human urine, serum, and plasma on the ARCHITECT c Systems and the AEROSET System. SUMMARY AND EXPLANATION OF TEST In addition to beverages, ethanol (ethyl alcohol or alcohol) can also be found in high concentrations in a variety of products such as mouthwashes, colognes, candies, and medicinal preparations. When alcohol is ingested, it will permeate all tissues of the body within one hour. About 95% of the alcohol is metabolized in the liver, and the remainder is excreted unchanged. Alcohol intoxication can lead to birth defects (e.g., fetal alcohol syndrome), loss of alertness, stupor, coma, and death. Determination of ethyl alcohol concentration is commonly used for measuring legal impairment, investigating forensic evidence, diagnosing and/or treating alcohol dependency, as well as detecting alcohol poisoning. Gas chromatography techniques and several enzymatic methods are available for determination of ethyl alcohol. 1,2 These techniques either require specimen pretreatment or require incubation periods ranging from 10 to 60 minutes. 3 Administration of pure alcohol to fasting men produced the following average maximal concentrations at the times indicated: Dose Concentration (mg/dl) Time (h) 0.5 ml/kg ml/kg ml/kg These concentrations decreased at an average rate of 18.9 mg/dl/h. At concentrations greater than 20 mg/dl, the rate of metabolism is independent of concentration, i.e., zero-order. 1 PRINCIPLES OF PROCEDURE MULTIGENT Ethanol assay is a liquid, ready-to-use, kinetic assay based on the high specificity of alcohol dehydrogenase (ADH) for ethyl alcohol. In the presence of ADH and nicotinamide adenine dinucleotide (NAD), ethanol is readily oxidized to acetaldehyde and NADH. ADH Ethyl alcohol + NAD Acetaldehyde + NADH The enzymatic reaction can be monitored spectrophotometrically at 340/412 nm (416 nm for c 4000 and c 16000). Methodology: Alcohol Dehydrogenase REAGENTS Reagent Kit 3L36-20 MULTIGENT Ethanol is supplied as a liquid, ready-to-use, two-reagent kit which contains: 2 x 14 ml 2 x 14 ml Estimated tests per kit are 200. Calculation is based on the minimum reagent fill volume per kit. Reactive Ingredients TRIS buffer Concentration mg/ml ADH 4.0 mg/ml NAD g/ml Inactive Ingredients: and contain sodium azide (< 0.1%) as a preservative. REAGENT HANDLING AND STORAGE Reagent Handling Ready for Use. Before use, invert several times, avoiding the formation of bubbles. Ready for Use. Before use, invert several times, avoiding the formation of bubbles. Remove air bubbles, if present in the reagent cartridge, with a new applicator stick. Alternatively, allow the reagent to sit at the appropriate storage temperature to allow the bubbles to dissipate. To minimize volume depletion, do not use a transfer pipette to remove the bubbles. CAUTION: Reagent bubbles may interfere with proper detection of reagent level in the cartridge, causing insufficient reagent aspiration that could impact results. When either the or the reagent cartridge becomes empty, replace both cartridges and validate the system by analyzing controls. Reagent Storage Reagent stability is 56 days if the reagent is uncapped and onboard. Unopened reagents are stable until the expiration date when stored at 2 to 8 C. Indications of Deterioration Instability or deterioration should be suspected if there are visible signs of leakage, extreme turbidity, microbial growth, if calibration does not meet the appropriate package insert and/or instrument-specific operations manual criteria, or if controls do not meet the appropriate criteria. WARNINGS AND PRECAUTIONS Precautions for Users For in vitro diagnostic use. Do not use components beyond the expiration date. Do not mix materials from different kit lot numbers. This product contains sodium azide; for a specific listing, refer to the REAGENTS section of this package insert. Contact with acids liberates very toxic gas. This material and its container must be disposed of in a safe way. NOTE: Refer to Section 8 of the instrument-specific operations manual for proper handling and disposal of reagents containing sodium azide. Ethanol is volatile. Specimens must be tightly capped to avoid evaporation. Do not leave calibrators, controls, or specimens uncapped longer than necessary. Store tightly capped inside a refrigerator to prevent evaporation of alcohol. CAUTION: This product requires the handling of human specimens. It is recommended that all human sourced materials are considered potentially infectious and be handled in accordance with the OSHA Standard on Bloodborne Pathogens. 4 Biosafety Level 2 5 or other appropriate biosafety practices 6,7 should be used for materials that contain or are suspected of containing infectious agents. 1

2 SPECIMEN COLLECTION AND HANDLING Serum, plasma, and urine can be used for this assay. Anticoagulants such as EDTA, citrate, fluoride/oxalate and heparin can be used for collection of plasma samples. Collect urine specimens in plastic or glass containers. Testing of fresh urine specimens is suggested. Samples within a ph range of 3 to 11 are suitable for testing with this assay. Do not use alcohol as a disinfectant when collecting or storing blood specimens. An effort should be made to keep pipetted samples free of gross debris. It is recommended that highly turbid specimens be centrifuged before analysis. Adulteration of the urine sample may cause erroneous results. If adulteration is suspected, obtain another sample and forward both specimens to the laboratory for testing. NOTE: Stored specimens must be inspected for particulates prior to testing. If present, mix and centrifuge the specimen to remove particulates. For total sample volume requirements, refer to Section 5 of the instrument-specific operations manual. Specimen Storage Serum/Plasma 8 Temperature Room Temperature (20 to 25 C) Refrigerated (4 to 8 C) Frozen (-20 C) Urine 8 Temperature Refrigerated (4 to 8 C) PROCEDURE Materials Provided 3L36-20 MULTIGENT Ethanol Materials Required but not Provided Products are ordered separately. Maximum Storage Up to two weeks Up to six months Up to six months Maximum Storage Up to 30 days 3L36-01 MULTIGENT Ethanol Neg Cal 1 x 5 ml 3L36-02 MULTIGENT Ethanol 100 Cal 1 x 5 ml 3L36-10 MULTIGENT Ethanol 50 Control 1 x 5 ml 3L36-12 MULTIGENT Ethanol 300 Control 1 x 5 ml Assay Procedure For a detailed description of how to run an assay on an ARCHITECT c System or the AEROSET System, refer to Section 5 of the instrument-specific operations manual. Specimen Dilution Procedure Samples exceeding mg/dl may be manually diluted with MULTIGENT Ethanol Neg Cal and rerun. The dilution must be performed so the diluted test results read greater than the assay sensitivity of 10.0 mg/dl. The operator must enter the manual dilution factor in the patient or control order screen. The system uses this dilution factor to automatically correct the concentration by multiplying the result by the entered factor. The printed result is the reportable result if no errors are present. NOTE: If the operator does not enter the manual dilution factor, the printed result must be multiplied by the manual dilution factor before reporting the result. (Volume of Sample + Volume of Dilution Reagent) Manual Dilution Factor = Volume of Sample For detailed information on ordering dilutions, refer to Section 5 of the instrument-specific operations manual. CALIBRATION Calibration is stable for approximately 13 days (312 hours) for MULTIGENT Ethanol reagent. Calibration is required with each change in reagent lot number. For information on calibrator and control standardization, refer to the MULTIGENT Ethanol Calibrators and Controls package insert. Both MULTIGENT Ethanol Neg Cal and the MULTIGENT Ethanol 100 Cal should be used to calibrate the assay. For a detailed description of how to calibrate an assay, refer to Section 6 of the instrument-specific operations manual. QUALITY CONTROL As appropriate, refer to your laboratory standard operating procedure(s) and/or quality assurance plan for additional quality control requirements and potential corrective actions. Two levels of controls are to be run every 24 hours of assay use. If more frequent control monitoring is required, follow the established quality control procedures for your laboratory. If quality control results do not meet the acceptance criteria defined by your laboratory, patient values may be suspect. Follow the established quality control procedures for your laboratory. Recalibration may be necessary. Review quality control results and acceptance criteria following a change of reagent or calibrator lot. RESULTS Results for the MULTIGENT Ethanol assay can be reported as mg/dl, mmol/l, or percentage (%). To convert results from mg/dl to mmol/l, multiply mg/dl by Serum/Plasma The rate of alcohol metabolism and excretion varies among individuals and is dependent upon factors such as gender, age, body weight, stomach content, concurrent use of medication, and health condition. The legal definition of intoxication varies. Urine Urine ethanol concentrations are often used to estimate blood concentrations. During the elimination phase, the urine/blood ratio of 1.3 applies and provides a valid estimate in most cases. 9 Jones has reviewed the use of urine as a specimen for ethanol analysis. 10 Refer to the instrument-specific operations manual for information on results calculations. ARCHITECT System Operations ManualAppendix C AEROSET System Operations ManualAppendix A Representative performance data are given in the SPECIFIC PERFORMANCE CHARACTERISTICS section of this package insert. Results in individual laboratories may vary. LIMITATIONS OF THE PROCEDURE Legal alcohol intoxication levels vary. The test result should be interpreted in light of clinical signs and symptoms. The test is designed for use with human urine, serum, and plasma only. Increased levels of lactic acid and LDH in postmortem samples may cause elevated ethyl alcohol results. It is possible that other substances and/or factors (e.g., technical or procedural errors) not listed in the Specificity table may interfere with the test and cause false results. 2

3 SPECIFIC PERFORMANCE CHARACTERISTICS Sensitivity Sensitivity, defined as the lowest concentration that can be differentiated from the negative calibrator with 99% confidence, is 10.0 mg/dl. Performance studies for MULTIGENT Ethanol produced a sensitivity of 1.4 mg/dl on an ARCHITECT c System or the AEROSET System. Method Comparison One hundred twenty-five clinical specimens were assayed for ethanol concentration by a commercially available assay (x) and by the Ethanol assay (y). A linear regression equation of y = 1.02x and a correlation coefficient (r) of were obtained. The MULTIGENT Ethanol assay was tested on an ARCHITECT c System (y) and on a commercially available clinical chemistry analyzer (x). Ninety-one clinical samples were assayed. A linear regression equation of y = 1.07x 0.32 and a correlation coefficient (r) of were obtained. Samples were also assayed on the AEROSET System (x) and the results were compared to the results from an ARCHITECT c System (y). A linear regression equation of y = 1.10x 1.15 and a correlation coefficient (r) of were obtained. Interference The following data were generated using the same reagent on a commercially available clinical chemistry analyzer. Grossly hemolyzed (800 mg/dl hemoglobin), icteric (30 mg/dl bilirubin), and lipemic (1,000 mg/dl triglycerides) samples were found to have no interference with the assay. Linearity The MULTIGENT Ethanol assay can accurately quantitate alcohol concentrations within a range of 10.0 mg/dl (0.01%) to mg/dl (0.6%). Precision The following table summarizes the precision results generated by repeat testing of the MULTIGENT Ethanol 100 Cal and MULTIGENT Ethanol 50 and 300 Controls: 6 reps x 2 runs x 5 days on the AEROSET System. Precision on an ARCHITECT c System and the AEROSET System is 5% total CV. BIBLIOGRAPHY 1. Baselt RC. Disposition of Toxic Drugs and Chemicals in Man, 5th ed. Foster City, CA: Chemical Toxicology Institute; 2000: Beutler HO: Ethanol. In: Bergmeyer HU, editor. Methods of Enzymatic Analysis, Vol. VI, 3rd ed. New York: Academic Press, 1984: Redetzki HM, Dees WL. Comparison of four kits for enzymatic determination of ethanol in blood. Clin Chem 1976:22, US Department of Labor, Occupational Safety and Health Administration. 29 CFR Part Bloodborne Pathogens. 5. US Department of Health and Human Services. Biosafety in Microbiological and Biomedical Laboratories, 5th ed. Washington, DC: US Government Printing Office, January World Health Organization. Laboratory Biosafety Manual, 3rd ed. Geneva: World Health Organization, Sewell DL, Bove KE, Callihan DR, et al. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved GuidelineThird Edition (M29-A3). Wayne, PA: Clinical and Laboratory Standards Institute, Guder WG, da Fonseca-Wollheim F, Heil W, et al. The Quality of Diagnostic Samples. Darmstadt, Germany: GIT Verlag; 2001:28, Heise HA. Concentrations of alcohol in samples of blood and urine taken at the same time. J For Sci 1967:12, Jones AW. Urine as a biological specimen for forensic analysis of alcohol and variability in the urine-to-blood relationship. Toxicol Rev 2006;25: TRADEMARKS The ARCHITECT c System family of instruments consists of c 4000, c 8000, and c Systems. AEROSET, ARCHITECT, c 4000, c 8000, c 16000, c System, and MULTIGENT are trademarks of Abbott Laboratories in various jurisdictions. All trademarks are property of their respective owner. Ethanol N Concentration Mean Total (mg/dl) SD %CV 50 Control Cal Control Specificity The following data were generated using the same reagent on a commercially available clinical chemistry analyzer. Various structurally related organic compounds were tested for cross reactivity in the assay. The following table summarizes the results: Compound Level Tested (mg/dl) % Cross Reactivity Acetaldehyde 2,000 0 Acetone 2,000 0 n-butanol 2, Ethylene glycol 2,000 0 Isopropanol 2,000 0 Methanol 2,000 0 n-propanol 2,

4 c Systems Assay Parameters Ethanol Configure assay parameters General General о Calibration о SmartWash о Results о Interpretation Assay: EtohM Type: Photometric Version: Number: 2847 Assay availability: Enabled Reaction definition о Reagent / Sample о Validity checks Reaction mode: Rate up Primary Secondary Read times Wavelength: 340 / Main: Last required read: 24 Absorbance range: Sample blank type: None Flex: Color correction: о Reaction definition Reagent / Sample о Validity checks R1 R2 Reagent: ETOHM Reagent volume: Diluent: None Water volume: Diluent dispense mode: Type 0 Dispense mode: Type 0 Type 0 Diluted Default Dilution name Sample sample Diluent Water Dilution factor dilution STANDARD : 12.0 = 1:1.00 о Reaction definition о Reagent / Sample Validity checks Reaction check: None Configure assay parameters SmartWash о General о Calibration SmartWash о Results о Interpretation Assay: EtohM COMPONENT REAGENT / ASSAY WASH Volume Replicates Cuvette Trig 10% Detergent B 345 Configure assay parameters Results о General о Calibration о SmartWash Results о Interpretation Assay: EtohM Assay number: 2847 Dilution default range: Result units: mg/dl Low-Linearity: 10.0 High-Linearity: Configure result units Assay: EtohM Version: Result units: mg/dl Decimal places: 1 [Range 0 4] Correlation factor: Intercept: Rate linearity %: Configure assay parameters Calibration о General Calibration о SmartWash о Results о Interpretation Assay: EtohM Calibration method: Linear Calibrators о Volumes о Intervals о Validity checks Calibrator set: Calibrator level: ( ) Concentration: ETH Blank: ETH1 (Ethanol Neg Cal) 0.0 Cal 1: ETH2 (Ethanol 100 Cal) Replicates: 2 [Range 1 3] о Calibrators Volumes о Intervals о Validity checks Calibrator: ETH Diluted Calibrator level Sample sample Diluent Water Blank: ETH Cal 1: ETH о Calibrators о Volumes Intervals о Validity checks Calibration intervals: Full interval: 312 (hours) Calibration type: Adjust type: None о Calibrators о Volumes о Intervals Validity checks Blank absorbance range: Span: Blank Blank Span absorbance range: Expected cal factor: 0.00 Expected cal factor tolerance %: 0 Due to differences in instrument systems and unit configurations, version numbers may vary. The c8000 Secondary Wavelength is 412 nm; the c4000 and c16000 Secondary Wavelength is 416 nm. Calibrator names added for clarification only and will not appear on the screen. Refer to concentration specified on calibrator labeling. In ARCHITECT software version 5.00 and above, these values are defined on the Configure calibrator set screen. The linear low value (Low-Linearity) is equal to the Sensitivity value in the reagent package insert. 4

5 Ethanol Assay Configuration: Outline Page Assay Name Assay # Line EtohM 847 * Quantitative Ranges Min Panic-L L-Reference-H Panic-H Max L-Linear Range-H Reference Ranges N/A Assay Configuration: Base Page Reaction Mode Wavelength-Prim/Sec Read time-main/flex Linearity% RATE UP 340 / / Sample Blank Test Blank Read Time Abs Window Abs Limits ( ) S.Vol DS.Vol D.Vol W.Vol Standard Rgt Name/Pos Dil Diluent: * Dil Type# 0 Rgt Name/Pos R.Vol W.Vol Type# Reagent 1 ETOHM41 * Reagent 2 ETOHM42 * Reaction Check Read Time A/B Range Minimum 1 1 / Factor/Intercept Decimal Places Units 1.0 / mg/dl Assay Configuration: Calibration Page Calib Mode Interval (H) Linear 312 Blank/Calib Replicates Extrapolation% Span Span Abs Range 2 / 2 0 BLK Sample S.Vol DS.Vol D.Vol W.Vol Blk Abs Range BLK ETHNC C1 ETH Cal Deviation 0.0 FAC Limit (%) 10 Assay Configuration: SmartWash Page Rgt Probe Reagent Wash Vol Cuvette Assay Name Wash Vol Sample Probe Wash Refer to Assay Configuration in Section 2 of the AEROSET System Operations Manual for information regarding assay parameters. * User defined or instrument defined. The linear low value (L-Linear Range) is equal to the Sensitivity value in the reagent package insert. Refer to concentration specified on calibrator labeling. 5

6 Symbols in Product Labeling Concentration Contents of kit Authorized Representative in the European Community Identifies products to be used together Ingredients ETHANOL 3L36-20 In vitro diagnostic medical device Batch code/lot number Reagent 1 Reagent 2 Catalog number/list number Sequence number Consult instructions for use Use by/expiration date Manufacturer Temperature limitation Customer Support United States: Canada: International: ABBOTT (English speaking customers) (French speaking customers) Call your local Abbott representative Microgenics Corporation Fremont Blvd. Fremont, CA USA Microgenics GmbH Spitalhofstrasse 94 D Passau Germany Tel: +49 (0) Distributed by: Abbott Laboratories, Inc. Abbott Park, IL USA 2009 July D 6