—, © Clinical Chemistry Pentra Albumin (BCG Method) (For quantitative estimation of Serum and Plasma Albumin) Intended Use Pentra Albumin reagent is intended for the quantitative in vitro diagnostic determination of albumin in serum plasma by colorimetry. Albumin measurements are used in the diagnosis and treatment of numerous diseases. involving primarily the liver or kidneys. Clinical Significance Albumin is the main component of plasmatic proteins. Its essential role is the maintenance of osmotic pressure, It also assures the fixation and transport of a large number of products. The albumin serum constitutes a predictive factor in the alteration of the transport of biliubine, calcium and hormones due to a deteriorated functioning of the liver and/or due to inflammations. A relative increase of plasmatic albumin is observed in states of dehydration. The decreases are the result of malnutrition, synthesis alteration (hepatic pathologies) or a serious loss of albumin by the organism (traumas, bums, haemorthages, diarthea, nephrotic syndromes and cancers), Method Colorimetric test for the quantitative determination of albumin in serum and plasma using Bromcresol Green dye-binding procedure. This method allows to measure out simply and quickly albumin, unlike electrophoresis or salt fractionation determinations which are not very convenient in laboratories. The principle of this test has been discovered by Klotz and Walker (1947) while they were studying the link between Bovine Serum Albumin and Bromeresol green. Rodkey, in the 19655, after having transposed his works fon the Human Serum Albumin had proposed a methodology in which the optical density (OD) variation ‘was directly proportional to the albumin concentration, But the OD of the reagent, which was too high, made this, determination out of reach of most spectrophotometers. Furthermore, interferences with globulin fractions could be responsible of albumin overestimations in the low concentration range with the initial methodology. Later, ew methodologies at differant pH faster time reading and the use of Brij35 has allowed the development of reliable, more specificand precise manual or automatic, ‘methods within the reach of many analysers, At pH 4.20, in succinate buffer and with a nonionic surfactant Br)35, the Bromeresol Green (BCG) fixes itself selectively o the albumin of the sample, producing a blue Colour which is measured at 628 nm. The intensity of the colouring is directly proportional to the albumin concentration Abumin + 866 =" bumin-B66 complex Reagent : BCG Reagent Succinate butter, pH4.2 75 mM Bromocresol green 0.14git Pentra Albumin should be used according to this Feagent notice. The manufacturer cannot guarantee its performance if used otherwise. Handling 1. Remove the cap of he cassette, 2. Ifpresent, remove foamy using aplastic pipette. 3. Place the cassette into the refrigerated Pentra C400 & PentraC200 reagent compartment. Linearity :6 gmial Reference Range’: Albumin:3.2-8.5gnvdl “It is recommended that each laboratory should establish its own normal range. Calibrator For calibration, use: Pentra Multical Control PontraN Control & PontraP Control Forinternal quality control, use: Each control should be assayed daily andlor after a calibration. The frequency of controls and the confidence intervals, should correspond to laboratory guidelines and country- specific directives. You should follow federal, state and local guidelines for testing quality control materials. The results must be within the range of the defined confidence limits. Each laboratory should establish a procedure to follow if the results exceed these confidence limits. Is required butnot provided Automated clinical chemistry analyzer: Pentra C400 & Penira C200, 1 Caliorator: Pentra Multical Controls: Pentra N Control & Pentra P Control Standard laboratory equipment. enmerial | Medial |@ Clinical Chemistry Pentra Albumin (BCG Method) (For quantitative estimation of Serum and Plasma Albumin) Storage and Stability Reagents, in unopened cassettes, are stable up to the expiry date on the label i stored beiween 2-8°C. Waste Management Please refer to local legal requirements. Accuracy and Precisior Repeatability (within-run precision) 3 specimens of low, medium and high concentration and 2 controls are tested 20 times. Mean Value maid ov Control specimen 4 4361 058 Control specimen 2 296 3.08 ‘Specimen T 25) aa ‘Specimen 2 4472 0.36 ‘Specimen 3 5.005 om Reproducibility (total precision) 2 specimens (low and medium levels) and 2 controls are tested in duplicate for 20 days. Mean Value gid ov Control specimen 1 279 1.98 Contral specimen 2 498 id ‘Specimen T 2307 125 ‘Specimen 2 4.738) 578 Correlation: 90 patient samples (serum and plasma) are correlated with a commercial reagent taken as reference. Y = 0.95 X + 0.02 (g/dL) with a correlation coefficient ro = 0.9888, Interferences: Haemoglobin: No significant influence is observed up to 300 mg/dL. Triglycerides: No significant influence is observed up to an Intralipid® concentration (represen- tative of lipemia) of 612.5 mid. Tolal Bilicubin:No significant influence is observed up to 36 mg/dl. Direct Bilirubin: No significant influence is observed up to 36 mg/dL. ‘Ampicilin has been found to seriously interfere with BCG. ‘methods, Reference 4, Thomas L, Clinical Laboratory Diagnostics, 1 ed Frankfurt: THBooks Verlagsgesellschant (1998):652- 653. 2. Klotz IM and Walker FM, J. Phys. Colloid. Chem. (1947) 51: 666. 3. Rodkey FL. Arch. Biochem. Biophys. (1964) 108: 510. 4, Rodkey FL, Clin, Chem, (1965) 11: 478. 5. Webster D. A study of the interaction of Bromeresol Green with isolated serum globulin fractions. Clin Chim. Acta (1974) §3:109-115. 6. Heandez 0, Murray Land Doumas B. Clin. Chem. (1967) 13: 701 7. Gustafsson Jan EC. Improved specificity of serum albumin determination and estimation of «Acute phase reactants» by use of the bromeresol green reaction, Clin, Chem. (1976) 22: 616-622. 8, Dow Dand Pinto PVC. Clin, Chem. (1969) 15: 1006. 9, Doumas BT, Watson WA and Biggs HG. Albumin standards and the measurement of serum albmin with bromocresol green. Clin. Chim. Acta (1971) 34 (1): 87-98, 10, Evaluation of Precision Performance of Clinical Chemistry Devices. Approved Guideline, CLS! (NCCLS) document EP5-A (1999) 19, ‘11. Evaluation of the Linearity of Quantitative Analytical Methods. Approved Guideline, CLSI (NCCLS) document EPE-A (2003) 23. 42, Method Comparison and Bias Estimation Using Patient Samples. Approved Guideline, 2n4 ed., CLS! (NCCLS) document EP9-A2 (2002) 22. Manufactured By HORIBA India Private Limited Plot No. 28, Sector-7, .LE, SIOCUL, Haridwar-248408, Utarakhand, Inia enmerial | Medial |