INSTRUCTIONS FOR USE

Transcription

1 INSTRUCTIONS FOR USE VITROS Chemistry Products TRIG Slides TRIG Triglyceride For Coatings 3400 and Above Rx ONLY Intended Use For in vitro diagnostic use only. VITROS Chemistry Products TRIG Slides quantitatively measure triglyceride (TRIG) concentration in serum and plasma using VITROS 250/350/950/5,1 FS and 4600 Chemistry Systems and the VITROS 5600 Integrated System. Triglyceride measurements are used in the diagnosis and treatment of patients with diabetes mellitus, nephrosis, liver obstruction, other diseases involving lipid metabolism, or various endocrine disorders. Summary and Explanation of the Test Triglycerides, fatty acid esters of glycerol, represent the major form of fat found in the body; their primary function is to store and provide cellular energy. The concentration of triglycerides in the plasma at any given time is a balance between the rates of entry and removal. Triglyceride concentrations in the plasma vary with age and gender. Moderate increases occur during growth and development. Triglycerides are used for the evaluation of hyperlipidemias; high concentrations may occur with hypothyroidism, nephrotic syndrome, glycogen storage diseases, and diabetes mellitus. Extremely high triglyceride concentrations are common in acute pancreatitis. 1 Principles of the Procedure The VITROS TRIG Slide method is performed using the VITROS TRIG Slides and the VITROS Chemistry Products Calibrator Kit 2 on VITROS 250/350/950/5,1 FS and 4600 Chemistry Systems and the VITROS 5600 Integrated System. The VITROS TRIG Slide is a multilayered, analytical element coated on a polyester support. The analysis is based on an enzymatic method as described by Spayd et al. 2 A drop of patient sample is deposited on the slide and is evenly distributed by the spreading layer to the underlying layers. The Triton X-100 surfactant in the spreading layer aids in dissociating the triglycerides from lipoprotein complexes present in the sample. The triglyceride molecules are then hydrolyzed by lipase to yield glycerol and fatty acids. Glycerol diffuses to the reagent layer, where it is phosphorylated by glycerol kinase in the presence of adenosine triphosphate (ATP). In the presence of L-α-glycerol-phosphate oxidase, L-α-glycerophosphate is then oxidized to dihydroxyacetone phosphate and hydrogen peroxide. The final reaction involves the oxidation of a leuco dye by hydrogen peroxide, catalyzed by peroxidase, to produce a dye. The density of the dye formed is proportional to the triglyceride concentration present in the sample and is measured by reflectance spectrophotometry. Test Type and Conditions Test Type Colorimetric VITROS System 5600, 4600, 5,1 FS, 950, 250/350 Approximate Incubation Time Temperature Wavelength Reaction Sample Volume 5 minutes 37 C (98.6 F) 540 nm 5.5 µl Not all products and systems are available in all countries. Version 2.0 Pub. No. J40115_EN 1 of 12

2 TRIG INSTRUCTIONS FOR USE Warnings and Precautions Reaction Scheme lipoproteins surfactant triglycerides + proteins triglycerides + H 2 O lipase glycerol + fatty acids glycerol + ATP glycerol kinase MgCl 2 L-α-glycerophosphate + ADP L-α-glycerophosphate + O 2 L-α-glycerol-phosphate oxidase dihydroxyacetone phosphate + H 2 O 2 H 2 O 2 + leuco dye peroxidase dye + 2H 2 O Warnings and Precautions For in vitro diagnostic use only. Reagents WARNING: Take care when handling materials and samples of human origin. Since no test method can offer complete assurance that infectious agents are absent, consider all clinical specimens, controls, and calibrators potentially infectious. Handle specimens, solid and liquid waste, and test components in accordance with local regulations and CLSI Guideline M29 3 or other published biohazard safety guidelines. For specific warnings and precautions for calibrators, quality control materials, and other components, refer to the Instructions for Use for the appropriate VITROS product, or to other manufacturer s product literature. Slide Diagram Slide Ingredients 1. Upper slide mount 2. Spreading layer (TiO 2 ) Reactive Ingredients per cm 2 Lipase (Pseudomonas sp., E.C ) 0.08 U; peroxidase (horseradish root, E.C ) 0.52 U; glycerol kinase (Cellulomonas sp., E.C ) 0.35 U; L-α-glycerophosphate oxidase (Pediococcus sp., E.C ) 0.19 U; Triton X mg; 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-bis(4- dimethylaminophenyl) imidazole (leuco dye) 0.04 mg; and adenosine triphosphate 0.14 mg. Other Ingredients Pigment, binders, buffer, surfactants, stabilizers, scavenger, enzyme cofactors, dye solubilizer and cross-linking agent. Reagent Handling Triton X-100 lipase 3. Reagent layer buffer, ph 8.0 glycerol kinase ATP L-α-glycerophosphate oxidase peroxidase leuco dye 4. Support Layer 5. Lower slide mount Caution: Do not use slide cartridges with damaged or incompletely sealed packaging. Inspect the packaging for signs of damage. Be careful when opening the outer packaging with a sharp instrument so as to avoid damage to the individual product packaging. Reagent Preparation IMPORTANT: The slide cartridge must reach room temperature, C (64 82 F), before it is unwrapped and loaded into the slide supply. 1. Remove the slide cartridges from storage. 2. Warm the wrapped cartridge at room temperature for 60 minutes. 3. Unwrap and load the cartridge into the slide supply. Note: Load the cartridges within 24 hours after they reach room temperature, C (64 82 F). 2 of 12 Pub. No. J40115_EN Version 2.0

3 INSTRUCTIONS FOR USE Reagent Storage and Stability TRIG Reagent Storage and Stability VITROS TRIG Slides are stable until the expiration date on the carton when they are stored and handled as specified. Do not use beyond the expiration date. Reagent Storage Condition Stability Unopened Frozen -18 C ( 0 F) Until expiration date Opened On-analyzer System turned on 1 week On-analyzer System turned off 2 hours Verify performance with quality control materials: If the system is turned off for more than 2 hours. After reloading cartridges that have been removed from the slide supply and stored for later use. Specimen Collection, Preparation and Storage Specimens Recommended Serum Plasma: Heparin (Lithium and Sodium) Serum is the specimen of choice because it is the basis for the US National Institutes of Health recommendations relating lipid levels with cardiac risk. Heparin plasma results have been reported as being within 1% of serum results. 4 IMPORTANT: Certain collection devices have been reported to affect other analytes and tests. 5 Owing to the variety of specimen collection devices available, Ortho-Clinical Diagnostics is unable to provide a definitive statement on the performance of its products with these devices. Confirm that your collection devices are compatible with this test. Specimens Not Recommended Plasma: EDTA 4 Serum and Plasma Specimen Collection and Preparation Collect specimens using standard laboratory procedures. 6, 7 Note: For details on minimum fill volume requirements, refer to the operating instructions for your system. Patient Preparation No special patient preparation is necessary. Special Precautions Equipment must be soap-free and glycerol-free. Do not use collection tubes with glycerol-lubricated stoppers. Centrifuge specimens and remove the serum or plasma from the cellular material within 4 hours of collection. 9 Specimen Handling and Storage Handle and store specimens in stoppered containers to avoid contamination and evaporation. Mix samples by gentle inversion and bring to room temperature, C (64 82 F), prior to analysis. Specimen Storage and Stability 9 Storage Temperature Stability Room Temperature C (64 82 F) 3 days Refrigerated 2 8 C (36 46 F) 7 days Frozen -18 C ( 0 F) 6 months IMPORTANT: Avoid repeated freeze-thaw cycles. Version 2.0 Pub. No. J40115_EN 3 of 12

4 TRIG INSTRUCTIONS FOR USE Testing Procedure Testing Procedure Materials Provided VITROS Chemistry Products TRIG Slides Materials Required but Not Provided VITROS Chemistry Products Calibrator Kit 2 Quality control materials, such as VITROS Chemistry Products Performance Verifier I and II VITROS Chemistry Products 7% BSA (manual dilution) VITROS Chemistry Products FS Diluent Pack 2 (BSA/Saline) [chamber D2 (BSA) for on-analyzer dilution] Operating Instructions Check reagent inventories at least daily to ensure that quantities are sufficient for the planned workload. For additional information, refer to the operating instructions for your system. IMPORTANT: Sample Dilution Bring all fluids and samples to room temperature, C (64 82 F), prior to analysis. Serum and Plasma If samples are grossly lipemic or show triglyceride concentrations that exceed the system s measuring (reportable or dynamic) range: Manual Sample Dilution 1. Dilute the sample with VITROS 7% BSA. 2. Reanalyze. 3. Multiply the results by the dilution factor to obtain an estimate of the original sample s triglyceride concentration. On-Analyzer Sample Dilution (VITROS 5600 Integrated, VITROS 5,1 FS/4600 and VITROS 250/350 Systems only) Refer to the operating instructions for your system for more information on the On-Analyzer Dilution Procedure. For VITROS 5600 Integrated and VITROS 5,1 FS/4600 Chemistry Systems, use VITROS Chemistry Products FS Diluent Pack 2 for the dilution. Calibration Required Calibrators VITROS Chemistry Products Calibrator Kit 2 Calibrator Preparation, Handling, and Storage Refer to the Instructions for Use for VITROS Calibrator Kit 2. Calibration Procedure Refer to the operating instructions for your system. When to Calibrate Calibrate: When the slide lot number changes. When critical system parts are replaced due to service or maintenance. When government regulations require. For example, in the USA, CLIA regulations require calibration or calibration verification at least once every six months. The VITROS TRIG test may also need to be calibrated: If quality control results are consistently outside acceptable range. After certain service procedures have been performed. For additional information, refer to the operating instructions for your system. Calculations Reflectance from the slide is measured at 540 nm after the fixed incubation time. Once a calibration has been performed for each slide lot, triglyceride concentration in unknown samples can be determined using the software-resident endpoint colorimetric math model and the response obtained from each unknown test slide. 4 of 12 Pub. No. J40115_EN Version 2.0

5 INSTRUCTIONS FOR USE Quality Control TRIG Validity of a Calibration Calibration parameters are automatically assessed by the system against a set of quality parameters detailed in the Coefficients and Limits screen on VITROS 250/350/950 Systems (on the VITROS 5600 Integrated and VITROS 5,1 FS/ 4600 Systems, see the Review Assay Data screen). Failure to meet any of the pre-defined quality parameters results in a failed calibration. The calibration report should be used in conjunction with quality control results to determine the validity of a calibration. Measuring (Reportable or Dynamic) Range Conventional Units SI Units Alternate Units (g/l) For out-of-range samples, refer to Sample Dilution. Traceability of Calibration Values assigned to the VITROS Chemistry Products Calibrator Kit 2 for triglyceride are traceable to the Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) The Ortho Clinical Diagnostics calibration laboratory uses SRM 1951 to calibrate a glycerol phosphate oxidase (GPO) triglyceride spectrophotometric method 11 to support triglyceride value assignment for VITROS Calibrator Kit 2. Quality Control Quality Control Material Selection IMPORTANT: VITROS Performance Verifiers are recommended for use with the VITROS Chemistry and Integrated Systems. Evaluate the performance of other commercial control fluids for compatibility with this test before using for quality control. Control materials other than VITROS Performance Verifiers may show a difference when compared with other triglyceride methods if they: Depart from a true human matrix. Contain high concentrations of preservatives, stabilizers, or other nonphysiological additives. Do not use control materials stabilized with ethylene glycol. Quality Control Procedure Recommendations Choose control levels that check the clinically relevant range. Analyze quality control materials in the same manner as patient samples, before or during patient sample processing. To verify system performance, analyze control materials: After calibration. According to local regulations or at least once each day that the test is being performed. After specified service procedures are performed. Refer to the operating instructions for your system. If control results fall outside your acceptable range, investigate the cause before deciding whether to report patient results. For general quality control recommendations, refer to Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline Third Edition 12 or other published guidelines. For additional information, refer to the operating instructions for your system. Quality Control Material Preparation, Handling, and Storage Refer to the Instructions for Use for VITROS Chemistry Products Performance Verifier I and II or to other manufacturer's product literature. Results Reporting Units and Unit Conversion The VITROS Chemistry and Integrated Systems may be programmed to report TRIG results in conventional, SI, and alternate units. Conventional Units SI Units Alternate Units mg/dl mmol/l (mg/dl ) g/l (mg/dl 0.01) Version 2.0 Pub. No. J40115_EN 5 of 12

6 TRIG INSTRUCTIONS FOR USE Limitations of the Procedure Limitations of the Procedure Known Interferences The VITROS Chemistry Products TRIG Slide method was screened for interfering substances following CLSI Protocol EP7-A2. 13 The substances listed in Tables 1 and 2, when tested in serum at the concentrations indicated, caused the bias shown. Interferent claims were cited whenever the maximum bias became equal to or exceeded the specificity requirement for TRIG. Materials exhibiting biases less than the specificity requirement have been cited as substances that do not interfere. It is possible that other interfering substances may be encountered. These results are representative; however, your results may differ somewhat due to test-to-test variation. The degree of interference at concentrations other than those listed might not be predictable. Table 1 Known Interferences: Lowest Interfering Concentration Levels Interferent Ethamsylate Hydroxyurea L-Dopa Lowest Interfering Concentration * Conv. SI Conv. Triglyceride Concentration SI Conv. Triglyceride Bias SI * The lowest interfering concentration of each substance was determined using the maximum 95% confidence interval of the quadratic regression that met or exceeded the specificity requirement (+/-12 mg/dl at triglyceride concentrations at or below 150 mg/dl and +/- 8% at triglyceride concentrations above 150 mg/dl). Table 2 Known Interferences: Interference at Upper Therapeutic Concentration Level Interferent Upper Therapeutic Concentration Conv. SI Ethamsylate Hydroxyurea L-Dopa Triglyceride Concentration Conv. SI Triglyceride Bias at Upper Therapeutic Concentration * Conv. SI * The bias is the maximum 95% confidence interval for the largest mean difference observed at the upper therapeutic concentration. Free glycerol 14 Free glycerol in serum is measured along with the glycerol from the hydrolysis of triglycerides and diglycerides. Certain clinical conditions show high endogenous free glycerol levels. Some drugs are also known to produce elevated glycerol levels in serum. Triglyceride results from samples of such patients will not reflect actual serum triglyceride content. Grossly lipemic samples show a slower rate of color development than do clear serums, which results in a negative bias. These samples often contain triglyceride concentrations greater than the system s measuring (reportable or dynamic) range. Grossly lipemic samples should be diluted prior to testing. For substances that were tested and did not interfere, refer to Specificity. Other Limitations Certain drugs and clinical conditions are known to alter triglyceride concentration in vivo. For additional information, refer to one of the published summaries. 15, 16 Expected Values Classification Triglyceride levels are categorized according to the classification scheme in the ATP III guidelines recommended by NCEP for samples collected from fasting patients. 8 6 of 12 Pub. No. J40115_EN Version 2.0

7 INSTRUCTIONS FOR USE Performance Characteristics TRIG Classification Conventional Units SI Units Alternate Units (g/l) Normal < 150 < 1.69 < 1.50 Borderline High High Very High Performance Characteristics Limit of Detection The limit of detection (LoD) for the VITROS Triglyceride test using delipidized human serum pools is 3.8 mg/dl, with proportions of false positives (α) less than 1% and false negatives (β) less than 1% based on 1100 determinations with 100 blanks and 1000 low-level samples. The Limit of Blank (LoB) is 2.0 mg/dl and the Limit of Quantitation (LoQ) is 3.8 mg/dl as determined by the lowest concentration at which precision and accuracy design requirements are met. The LoB, LoD and LoQ were verified consistent with NCCLS document EP17-A 10. The data presented are a representation of product performance. LoB * LoD ** LoQ *** * Limit of Blank, or the highest value likely to be observed with a sample containing no analyte with 95% level of confidence that it does not contain the analyte of interest. ** Limit of Detection, The minimum amount of analyte whose presence can be detected with 99% level of confidence under defined conditions. *** Limit of Quantitation, The minimum amount of analyte whose presence can be quantitatively determined with stated acceptable precision and trueness. Method Comparison The plots and data below show the results of a method comparison study with serum samples analyzed on the VITROS 5,1 FS Chemistry System and an Enzymatic Total Glycerol method traceable to Certified NIST (National Institute of Standards and Technology) Reference Material, SRM (Standard Reference Material) 1951, based on Clinical Laboratory Standards Institute (CLSI) Protocol EP9-A2 18 The table also shows the results of comparisons with serum samples on the VITROS 950 Chemistry System, the VITROS 350 Chemistry System, and the VITROS 5600 Integrated System against the VITROS 5,1 FS Chemistry System. This testing followed CLSI Protocol EP9-A2. 18 Conventional Units SI Units VITROS 5,1 FS Chemistry System VITROS 5,1 FS Chemistry System Comparative Method: Enzymatic Total Glycerol Comparative Method: Enzymatic Total Glycerol Version 2.0 Pub. No. J40115_EN 7 of 12

8 TRIG INSTRUCTIONS FOR USE Performance Characteristics System n Slope Correlation Coefficient Conventional Units Range of Sample Conc. Intercept Sy.x SI Units Range of Sample Conc. Intercept Sy.x VITROS 5,1 FS vs. comparative method * VITROS 950 vs. VITROS 5,1 FS VITROS 350 vs. VITROS 5,1 FS VITROS 5600 vs. VITROS 5,1 FS * Enzymatic Total Glycerol Analytical processing hardware and software algorithms on the VITROS 4600 Chemistry System are designed to the same specifications as those applied to the VITROS 5,1 FS Chemistry System. Assay performance on the VITROS 4600 System has been demonstrated to be comparable to that on the VITROS 5,1 FS System. All performance characteristics for VITROS 5,1 FS System are therefore applicable to the VITROS 4600 System. Precision Precision was evaluated with quality control materials on VITROS 350, 950, 5,1 FS Chemistry Systems and the VITROS 5600 Integrated System following NCCLS protocol EP5-A2 20 The data presented are a representation of test performance and are provided as a guideline. Variables such as sample handling and storage, reagent handling and storage, laboratory environment, and system maintenance can affect reproducibility of assay results. System VITROS 350 VITROS 950 VITROS 5,1 FS VITROS 5600 Specificity Conventional Units Mean Conc. Within Day SD * Within Lab SD ** Mean Conc. SI Units Within Day SD * Within Lab SD ** Within Lab CV% ** No. Observ. No. Days * Within Day precision was determined using two runs per day with two replications per run. ** Within Lab precision was determined using a single lot of slides and calibrating weekly. Analytical processing hardware and software algorithms on the VITROS 4600 Chemistry System are designed to the same specifications as those applied to the VITROS 5,1 FS Chemistry System. Assay performance on the VITROS 4600 System has been demonstrated to be comparable to that on the VITROS 5,1 FS System. All performance characteristics for VITROS 5,1 FS System are therefore applicable to the VITROS 4600 System. Substances that Do Not Interfere The substances listed in the table were tested with the VITROS TRIG Slides according to CLSI Protocol EP7-A2, 13 and found not to interfere at the concentrations shown. The substances were tested at a triglyceride concentration of approximately 150 mg/dl (1.69 mmol/l) and found not to interfere, bias <12 mg/dl (0.14 mmol/l). The substances were also tested at a triglyceride concentration of approximately 500 mg/dl (5.65 mmol/l) and found not to interfere, bias <8%. Compound Concentration Acetaminophen mg/dl 1324 µmol/l Acetylsalicylic acid 65.2 mg/dl 3.62 mmol/l Alprazolam 0.2 mg/dl 6.48 µmol/l Para-Aminosalicylic acid 79.4 mg/dl 5.22 mmol/l Amitriptyline µg/ml 3.61 µmol/l Amlodipine 0.01 mg/dl 245 nmol/l 8 of 12 Pub. No. J40115_EN Version 2.0

9 INSTRUCTIONS FOR USE Performance Characteristics TRIG Compound Concentration Amoxicillin 7.53 mg/dl 206 µmol/l Ascorbic acid 6.02 mg/dl 342 µmol/l Atorvastatin 600 µg/l 519 nmol/l Azithromycin 1.15 mg/dl 15.3 µmol/l Bilirubin, conjugated 38.9 mg/dl 462 µmol/l Bilirubin, unconjugated 27 mg/dl 462 µmol/l Carbenicillin 200 mg/dl 5.31 mmol/l Cephalexin 11.7 mg/dl 337 µmol/l Cholesterol * 503 mg/dl 13 mmol/l Ciprofloxacin 1 mg/dl 30.2 µmol/l Clonidine 0.01 µg/ml 43.5 nmol/l Clopidogrel 18 mg/dl 560 µmol/l Cystine 7 mg/dl 291 µmol/l Diatrizoate, sodium (Hypaque ) 521 mg/dl 8.2 mmol/l Dipyrone 120 mg/dl 3.6 mmol/l Dopamine µg/ml 5.87 µmol/l Estradiol 1.2 ng/ml 4.41 nmol/l Ethanol 400 mg/dl 86.8 mmol/l Fenofibrate 4.5 mg/dl 125 µmol/l Furosemide 6 mg/dl 181 µmol/l Gentisic acid 1.79 mg/dl 117 µmol/l Glutathione 92 mg/dl 3 mmol/l Glyburide 1.92 µg/ml 3.89 µmol/l Hemoglobin 600 mg/dl 6 g/l Hydrochlorothiazide 0.6 mg/dl 20.2 µmol/l Hydrocodone 0.02 mg/dl 0.67 µmol/l Ibuprofen mg/dl 2425 µmol/l Insulin 300 µiu/ml pmol/l Isoniazid 4 mg/dl 292 µmol/l Levothyroxine 0.1 mg/dl 1.29 µmol/l Lisinopril 0.03 mg/dl 0.74 µmol/l Metformin 4 mg/dl 310 µmol/l Methicillin sodium mg/dl 597 µmol/l Methimazole 1.2 mg/dl 105 µmol/l Methotrexate 91 mg/dl 2 mmol/l Methyldopa 1.5 mg/dl 71 µmol/l Metoprolol 0.5 mg/dl 18.7 µmol/l Naproxen mg/dl 2170 µmol/l Niacin 40 mg/dl 3.25 mmol/l Omega-3 Fatty Acid, DHA (docosahexaenoic acid) 90 mg/dl 2.74 mmol/l Omega-3 Fatty Acid, EPA (eicosapentaenoic acid) mg/dl 3.69 mmol/l Omeprazole 0.6 mg/dl 17.4 µmol/l Phospholipids 852 mg/dl 11.1 mmol/l Pioglitazone 2.7 mg/dl 76 µmol/l Prednisone 0.03 mg/dl 0.84 µmol/l Rifampin 6.43 mg/dl 78.1 µmol/l Sertraline 0.06 mg/dl 1.96 µmol/l Sitagliptin 5.22 mg/dl µmol/l Terazosin 3.02 µg/ml 7.8 µmol/l Total protein 10 g/dl 100 g/l Triamterene 0.89 mg/dl 35 µmol/l Version 2.0 Pub. No. J40115_EN 9 of 12

10 TRIG INSTRUCTIONS FOR USE References Compound Concentration L-Tyrosine mg/dl 4000 µmol/l Warfarin µg/ml 32.5 µmol/l * Cholesterol was evaluated using patient samples with TRIG concentrations of mg/dl ( mmol/l) References 1. Tietz NW (ed). Fundamentals of Clinical Chemistry. ed. 3. Philadelphia: WB Saunders; ; Spayd R, et al. Multilayer Film Elements for Clinical Analysis. Clin. Chem. 24: ; CLSI. Protection of Laboratory Workers from Occupationally Acquired Infections; Approved Guideline Third Edition. CLSI document M29-A3 (ISBN ). CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; NCEP. Recommendations for improving cholesterol measurement. A report from the Laboratory Standardization Panel of the National Cholesterol Education Program. NIH publication no :26 27; Calam RR. Specimen Processing Separator Gels: An Update. J Clin Immunoassay. 11:86-90; CLSI. Procedures for the Collection of Diagnostic Blood Specimens by Venipuncture; Approved Standard Sixth Edition. CLSI document H3-A6 (ISBN ). CLSI, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania USA; NCCLS. Procedures and Devices for the Collection of Diagnostic Capillary Blood Specimens; Approved Standard Fifth Edition. NCCLS document H4-A5 [ISBN ]. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA, NCEP. Third Report of the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III), Final Report. NIH Publication No : II-7, III-6. National Institutes of Health. Bethesda, Maryland: September Clinical Laboratory Handbook for Patient Preparation and Specimen Handling. Fascicle VI: Chemistry/Clinical Microscopy. Northfield, IL: College of American Pathologists; NCCLS. Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline. NCCLS document EP17-A [ISBN ]. NCCLS, 940 West Valley Road, Suite 1400 Wayne, Pennsylvania USA, Fossati P, Prencipe L. Serum Triglycerides Determined Colorimetrically with an Enzyme that Produces Hydrogen Peroxide. Clin. Chem. 28: ; CLSI. Statistical Quality Control for Quantitative Measurements: Principles and Definitions; Approved Guideline Third Edition. CLSI document C24-A3 (ISBN ). CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; CLSI. Interference Testing in Clinical Chemistry; Approved Guideline Second Edition. CLSI document EP7-A2 (ISBN ), CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; Stein EA, et al. National Cholesterol Education Program Recommendations for Triglyceride Measurement: Executive Summary. Clin. Chem. 41: ; Young DS. Effects of Drugs on Clinical Laboratory Tests. ed. 4. Washington D.C.: AACC Press; Friedman RB, Young DS. Effects of Disease on Clinical Laboratory Tests. Washington, D.C.: AACC Press; NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline. NCCLS Document EP9. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; NCCLS. Method Comparison and Bias Estimation Using Patient Samples; Approved Guideline Second Edition (Interim Revision). CLSI document EP9-A2-IR [ISBN ]. CLSI, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania USA; NCCLS. User Evaluation of Precision Performance with Clinical Chemistry Devices. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; NCCLS. Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline Second Edition. NCCLS document EP5-A2 [ISBN ]. CLSI, 940 West Valley Road, Suite 1400, Wayne, PA USA; of 12 Pub. No. J40115_EN Version 2.0

11 INSTRUCTIONS FOR USE Glossary of Symbols TRIG Glossary of Symbols Revision History Date of Revision Version Description of Technical Changes* Prescription Use statement added. Updated EC Representative address. Added USA to legal manufacture address First release of document * The change bars indicate the position of a technical amendment to the text with respect to the previous version of the document. When this Instructions For Use is replaced, sign and date below and retain as specified by local regulations or laboratory policies, as appropriate. Signature Obsolete Date Version 2.0 Pub. No. J40115_EN 11 of 12

12 TRIG INSTRUCTIONS FOR USE Revision History Ortho-Clinical Diagnostics Felindre Meadows Pencoed Bridgend CF35 5PZ United Kingdom Ortho-Clinical Diagnostics, Inc. 100 Indigo Creek Drive Rochester, NY USA VITROS is a registered trademark of Ortho-Clinical Diagnostics, Inc. Ortho-Clinical Diagnostics, Inc., of 12 Pub. No. J40115_EN Version 2.0