UMA Co., Ltd. provides a reagent for the determination of Total Bilirubin (T-Bil) concentration in serum or plasma using an enzymatic method. The product is designed for in vitro diagnostics and is compatible with various automated analyzers, with specific instructions for sample preparation and assay procedures. The expected normal reference range for T-Bil is 0.2 to 1.2 mg/dL, and the reagent must be stored properly to maintain stability and accuracy.
UMA Co., Ltd. provides a reagent for the determination of Total Bilirubin (T-Bil) concentration in serum or plasma using an enzymatic method. The product is designed for in vitro diagnostics and is compatible with various automated analyzers, with specific instructions for sample preparation and assay procedures. The expected normal reference range for T-Bil is 0.2 to 1.2 mg/dL, and the reagent must be stored properly to maintain stability and accuracy.
UMA Co., Ltd. provides a reagent for the determination of Total Bilirubin (T-Bil) concentration in serum or plasma using an enzymatic method. The product is designed for in vitro diagnostics and is compatible with various automated analyzers, with specific instructions for sample preparation and assay procedures. The expected normal reference range for T-Bil is 0.2 to 1.2 mg/dL, and the reagent must be stored properly to maintain stability and accuracy.
UMA Co., Ltd. provides a reagent for the determination of Total Bilirubin (T-Bil) concentration in serum or plasma using an enzymatic method. The product is designed for in vitro diagnostics and is compatible with various automated analyzers, with specific instructions for sample preparation and assay procedures. The expected normal reference range for T-Bil is 0.2 to 1.2 mg/dL, and the reagent must be stored properly to maintain stability and accuracy.
MEASURE T-BIL 2-19-6 Yokosuka Reagent for determination of Total Billirubin Matsudo, Chiba, Japan Enzymatic Method
2 ~ 8 °C IVD In vitro Diagnostics
QUALITY MANAGEMENT SYSTEM (BY TUV) = ISO 13485:2016 = DO NOT freeze 18 months/block from light
1. PURPOSE OF USE Reagent R-2 is ready for use
Providing a quantitative in vitro assay for the total Billirubin - Once open, Reagent stored on board the instrument is (T-Bil) concentration in serum or plasma. stable for 30 days with Hitachi 7180 Analyzers. - Applicable to various automated analyzers. 2. GENERAL INSTRUCTION a. For in vitro diagnostics use only. - Calibrator (separately sold): Put 2 mL of purified water to
b. Diagnosis should be made in a comprehensive the vials of Calibrator (Bili STD), leave at room
manner, in accordance with other related test results temperature for 20 minutes and sometimes gently invert
and clinical symptoms by the doctor in attendance. the vial before use. After reconstituting, Calibrator can be
c. For guaranteed results, usage of this product must used without dilution.
comply with the instruction in this manual. - Controls MEASURE Human Lyo L-1 and MEASURE Human Lyo L-2 (separately sold): Put 5 mL of purified d. If you use automatic analyzers, follow their water to the vials of controls (Lyo L-1 and Lyo L-2); leave instructions carefully. SUMMARY at room temperature for 45 minutes and sometimes gently
As discussed with normal metabolism, bilirubin is a invert the vial before use. After reconstituting, Controls
product of heme breakdown. It exists in conjugated (water can be used without dilution.
soluble) and unconjugated (lipid soluble) forms, which are 5. SAMPLE PREPARATION & STORAGE reported imprecisely as the direct and indirect fractions, - Serum: Wait until sample completely coagulated. Take respectively. Serum bilirubin is usually less than 1 mg/dL the supernatant to use as specimen. and primarily unconjugated. Elevated serum levels occur - Plasma: Treat blood sample by anticoagulant (Li-heparin in most significant liver diseases; degree of elevation and K2, K3-EDTA); leave it to stand for 3 hours or correlates with prognosis in primary biliary cirrhosis, centrifuge at 2000 rpm for 2 minutes; take the plasma alcoholic hepatitis, and fulminant liver failure. The layer (supernatant) and use as specimen. appearance of conjugated bilirubin in the blood is thought - Bilirubin is sensitive to light, avoid expose sample to light to be caused by reflux from the hepatocyte, but this does after collection. not discriminate between obstructive and parenchymal - AVOID use hemolytic sample. causes. Other causes of elevated bilirubin include - Analyze sample soon after collection. Gilbert's syndrome, increased production (e.g., hemolysis, - Stability: ineffective erythropoiesis, hematoma resorption), and 8 hours at 15-25°C inherited disorders of bilirubin transport. 3 days at 2-8°C
3. MATERIALS REQUIRED BUT NOT INCLUDED >30 days at (-60) – (-80)°C
- Saline 0.9 % and high grade purified water 6. MEASUREMENT PRINCIPLE
- Micropipet and other basic laboratory equipment. Total Bilirubin is oxidized by Bilirubin oxidase (BOD) and - Calibrators and Controls (separatedly sold) absorbance of Bilirubin will decrease. Total Bilirubin quantity can be obtained by measuring 4. REAGENT COMPOSITION & PREPARATION decreased absorbance. - Reagent R-1: Cholic acid; Ascorbate oxidase 𝐵𝑂𝐷 Reagent R-1 is ready for use Total Bilirubin + 1/2 O2 →−−−−−→ Biliverdin + H2O
7. ASSAY PROCEDURE Different measuring principle (n = 208) This product is compatible with various types of clinical Regression equation: y = 1.2202x - 0.1099 analyzer. An example of the assay procedure is indicated Correlation coefficient: r = 0.9893 below. (y: value obtained from this method) o Reference Materials for Calibration Sample + Reagent 1 37C Reagent 2 265s - NIST (SRM916) 5.0µL 135µL 45µL 10. EXPECTED VALUES o o 3 7C Meas. Abs I 3 7C Meas. Abs II Normal referance range 18s 317 s 0.2 ~ 1.2 mg/dL (3.4 ~ 20.5 µmol/L) (450/600nm) (450/600nm) Reference range should be established at each facility
∆Abs at 450/600nm Concentration and judgement should base on measurement results in a comprehensive manner together with clinical symptoms Perform the assay according to the instructions for and other measurement results. operating the automated analyzer Hitachi models. Refer 11. INTERFERENCES to the 13. INFORMATION FOR AUTOANALYZERS for - Hemolysis: No significant interference of hemoglobin the details of the assay method. Contact JAPAN concentration up to 20 mg/dL KANPEKI., JSC. For information about the - Lipemic: No significant interference of triglycerides parameters for other automated analyzers. concentration up to 3000 FTU 8. CALCULATION & UNIT CONVERSION - Ascorbic Acid: No significant interference of ascorbic Calculation acid concentration up to 50 mg/dL - Calculate ∆Abs of specimen & standards vs blank - For diagnostic purposes, the results should always be - Plot a calibration curve T-Bil (mg/dL) = f(∆Abs) assessed in conjunction with the patient’s medical history, - Calculate T-Bil concentration in specimen using the curve clinical examination and other findings. Please use (doing same procedure for Controls) another methods if the result is affected by any factors. Unit conversion 12. HANDLING, USAGE & DISPOSAL mg/dL x 17.1 = µmol/L Handling 9. PERFORMANCE & CORRELATION TEST 1. Specimen can be potentially positive for infectious a. Measuring range agents including hepatitis B virus and HIV. Wear glove and - The test is linear within a concentration range of 0 ~ 30 goggle when needed. mg/dL. (0 ~ 513 μmol/L). 2. In case reagents got into skin, eye or mouth by mistake, - If the concentration of sample exceeds assay range, wash it immediately with plenty of water and consult the dilute the sample with saline and repeat the measurement. doctor if needed. b. Lower Detection Limit: 0 mg/dL 3. If reagents are spilled, dilute with water and wipe it out. The lowest detectable level represents the lowest If specimen is spilled, spray 80% of alcohol over the measurable level of T-Bil that can be distinguished from specimen and wipe it out. zero. It is calculated as the absolute mean plus three Usage standard deviations of 20 replicates of an analysis free 1. Store reagents under specified condition. Do not use sample. after expiration date. c. Performance 2. Do not use the container and auxiliaries included in this - Sensitivity: Change in absorbance when measuring kit for other purposes. purified water as sample ranges from 0.001 to 0.05 and 3. Do not mix reagents of different lot for use. when measuring standard solution (total bilirubin 10 mg/dL) 4. Do not add to the reagent being used even if it is the as sample ranges from -0.25 to -0.04 absorbance unit. same lot number. - Specificity: The accuracy is within ±15.0%. Disposal - Reproducibility: CV value < 5.0%. 1. All specimens, as well as all instruments (e.g. test tubes) d. Correlation Test Results that come in contact with the specimens, must be treated
2/3 Revision 09/2022
by the following methods, or they must be treated 16. REFERENCES according to the manual for infectious medical waste 1. Anahat Dhillon MD, Randolph H. Steadman MD, in provided in each facility. Anesthesia and Uncommon Diseases (Sixth Edition), ◼ Sterilize with an autoclave, subjecting them to high 2012 pressure saturated steam at 121 °C for more than 20 2. In house data, UMA Diagnostics. minutes. Do not process waste containing sodium 17. MANUFACTURER hypochlorite solution with an autoclave. UMA Co., Ltd. ◼ Immerse at least one hour in sodium hypochlorite 2-19-6 Yokosuka, Matsudo City, Chiba solution (active chloride concentration of over 1000 ppm). Prefecture 270-0031 2. This reagent contains sodium azide. Sodium azide can TEL: 047-710-4871 (dial-in) FAX: 047-710-4872 react with lead pipe and/or steel pipe and can generate explosive metal azide. Make sure to use plenty of water at disposal. Concentration of sodium azide in R-2 is 0.05%.
13. INFORMATION FOR AUTOANALYZERS
For Hitachi Model Calculation Method 2-point end (Endpoint with Blank)
Temperature 37°C
Specimen 5 Volume (μL) R1 135 R2 45 Main 450 Wavelength (nm) Sub- 600 Point 1 10 Measurement Point 2 16 (cycle) Point 3 34
Calibration type Linear
Unit mg/dL
14. OTHER INSTRUCTIONS AND CAUTION
- Results may differ depending on the sample/reagent ratio. Adjust parameters for different analyzer. - Prepare the calibration curve on the day of determination.
