INTRODUCTION 1, Intinite Liquid Taglycerides is a reagent set for determination of triglycerides, based on enzymatic method using Lipoprotein lipase, Gilycerolkinase, Glycerol phasphats oxidaseand Peroxidase, Inttinite Liquid Teglycerdes sa ready-to-use reagent. Triglycerides can be determined in just 10 minutes at 37°C or 20 ‘minutes at AT. (25°C-20°C). Inttinite Liquid Tealycerdes is tinear upto 800 m9%. 4Intnnite Liquid Tiglycerides can be used on any Colorimeter, Spectrophotometer, Discrete semiautomated and Automated analyzer. Programme can be designed for any specific analyzer upon request 6, Tho influence of lipids, haemolysis and bilrubin (upto 8 mgx) Ie negiible. PRINCIPLE Caiyeeral released trom hydrolysis of Wighcerdes by Hpoprotein lipase le converted by glycerol kinase into glyceral -3- phosphate which is oxidsed by glycerol phosphate oxidase to chydroxyacetone phosphate and hydrogen peroxide. In presence of peroxidase, hydrogen peroxide oxidizes phenalc chromogen fo @ red coloured compound. LPL: Triglycerides > Fatty acids + Glycerol lycerot+arp___°%" _. Gtycerol-s-phosphate + ADP GPot Glycerel-s-phosphate——S"°” +, pinydroxyacetone phosphate , +H,0, H,0, + Phenolic chromogen POP” _. Red colour compound “Abbreviations LPL = Lipoprotein pase GPO. GK = Glycerol kinase POD ‘aycerol phosphate oxidase Peroxidase REAGENT STORAGE, STABILITY & HANDLING The kt should be stored at 26°C and is tablet the expiry date indicated ontnelabel ‘The reagent and standard are ready-to-use and are stable til expiry, when stored.a!2-€°C, DO NOT FREEZE THE REAGENT. ‘Tho reagont should be stored ony inthe amber bottle provided to protect from direct light. tore use Swi inthe roagont gantly. DO NOT SHAKE VIGOROUSLY. overtime, the reagent may develop alight pink colour This s expected and does not affect the reagent performance. Discard the reagent if the absorbance of the same exceeds 0.300 OD. against distiled water at 510nm Contamination of the reagent should be strctly avoided. Should the reagent develop turbicty 4 scardthe reagent COMPONENTS & CONCENTRATION OF WORKING SOLUTION Component Concentration + Butter pH 72 50 mmoll + Lipase 2000 IU + Glycerol kinase 2300 UI + Giycorel phosphate oxdase >1090 1Un + Peroxidase 500 IU + AIP + mmotl + Chromogen 2 mmol + Activators & stabiizers SPECIMEN COLLECTION & PRESERVATION Blood should be collected in a clean dry container. Avoid the use of plastic (oF silcorized container which may prolong clotting time. Serum or plasma should be separated from the cells at the earliest possible (within 30 minutes). For plasma collection folowing anticoagulants may be used: + EDTA 2 malml of blood * ciTRATE: 6 mgmt of Blood + HEPARIN 200. Wim of blood ‘Avoid use of Oxala ‘and Sodium Fluoride as anticoagulant. Tiglycerdes are stable for 4 days in neatly separated sorum or plasma a2, PROCEDURE . Ené-Point 10 mins, at 37°C/20 mins, at RT. (25 30°C) 5 Wavelength, 510 nm. (500 - 580 nm.) 1 Zero setting with Reagent Blank 1D. Blank absorbance limit <0.200 Abs. Sample volume 0.01 mi (1041) Reagent volume. 10m! Standard concentration 200 mer 1 Lineerty 800 mst Manual assay procedure Prewarm at room temperature (25-30°C) the required amount of reagent before use. Perform the assay as given below 1.0 ml procedure Serum /Plasma Standard oot mt = [Link] Reagent 10 mt 10 mi t.0 mt Incubation Incubate the assay mixture for 10 minutes at 37°C or 20 minutes at RIT. (25-30°C), After incubation, measure the absorbance against blank at 510 nm, (600-530 nm, Final colours stable for 30 minutes it not exposed to direct ight. Calculat © With standard 8 Absorbance of Sample 3 Cone. (mgi) = ane’ ST PAMPS _ 200 5 ‘Absorbance of Standard 8 © Wit factor for wavelength Cone. (mg) nge : 500 - 510 nm, 48 x Abs ance of SampleNOTE: 41. The specimen to reagent ratio can be altered proportionally without atfectng the results 2, Falsely elevated results are obtained due to lipemic samples. The Intfinite\iquid Tigycerides overcomes falsely elevated result with Use of Lipid clearing agent in the reagent formulation. Lipid clearing agent clears up the tubidly caused by lipemic same. EXPECTED VALUES Upto 170 mgr NOTE: Expected range varies from population to population therefore each laboratory should establish ts own normal range PROCEDURE LIMITATIONS 1. the tigycerdas value exceeds 800 mg% dilute the specimen with saline(1:1 ratio) and repeat the assay. Multiply the result obtained by 2 togettnecorrecttriglycerides value, Giycerol contamination n glasswareleads to erroneous results Care should be taken not to touch reagent and samples wit fingers, especially ater applying hand tion which may contain glycerin. QUALITY CONTROL ‘To ensure adequate quality conta, it i recommended that each batch should include a normal and an abnotmal commercial reference contro! serum. I should be realised that the use of qualty contol material checks both insttument and reagent functions together Factors which might affect the performance ofthis test include proper instrument furction, temperature Cont, cleaniness of glassware and accuracy of pipetting REFERENCES 1, Foosati,P,etalClin. Chem. 28,2077 (1982) 2. Honry, J. B., Clinleal Diagnosis and Management by Laboratory ‘Methods, 16th ed, W.S. Saunders, Phiadelphia, 1991, 0.208.211 3, Tietz, N.W,, Clinical Guide to Laboratory Tests, 2° od, WB, Saunders, Philadelphia, 1996, p. 1073-1091 4. Young, 0... 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