Cholesterol and LDL Examination - Edubirdie

University:
Massachusetts Institute of Technology
Course:
HST 198 | Health Sciences and Technology
Academic year:

2024
Views:

265

Pages:

2
Author:

mason mc44

CHOLESTEROL AND LDL EXAMINATION BLOOD Method for measuring LDL cholesterol levels: a. Zak method In plasma, cholesterol is bound in lipoprotein micelles. Plasma proteins precipitated with alcohol, the lipid fraction is extracted with acetone or ether. Residue what is difficult to evaporate is cholesterol. The cholesterol residue is dissolved with glacial acetic acid and stained with FeCl3 in sulfuric acid. b. Lieberman-Bourchard method This method does not carry out deprotenusation, the basis of the color formation reaction green-blue cholesterol with coloring reagent (a mixture of glacial acetic acid/anhydride 60/40). c. Enzymatic method This method produces more accurate results, only the reagents must be stored properly good because enzymes are easily damaged. 1) Testing principle LDL will be precipitated by heparin and sodium citrate. HDL and VLDL will obtained in the supernatant after centrifugation. Concentration of LDL cholesterol calculated as the difference of total cholesterol and cholesterol in the supernatant. 2) Calculation of LDL cholesterol levels Tools and materials - Human blood - Sterile lancet / syringe 2 ml, 70% alcohol cotton swab - Heparin and sodium citrate solution - Test tubes, micro tubes - Micro centrifuge, clinical photometer Work procedures ØBlood collection Approximately 1 ml of blood is taken, then the blood is centrifuged rotation at 4000 rpm for 15 minutes to obtain blood serum. ØMeasurement of cholesterol levels - Total cholesterol Take 10 µl of serum, then mix with as much enzyme reagent ("kit") 1000 µl, then vortexed and incubated for 10 minutes at temperature 20 – 25o C or 5 minutes at 37 o C then read with a photometer clinical. - Cholesterol in the supernatant • Take 100 µl of serum, put it in an Eppendrof tube then mixed with 1000 µl of precipitating reagent consisting of the mixture heparin and sodium citrate solution. • The mixture of serum and reagent is vortexed and then incubated for 10 minutes at a temperature of 20 – 25 o C or 5 minutes at temperature 37 o C and centrifuge. • After centrifuging, leave for 1 hour so that the LDL settles. • Take 100 µl of supernatant, put it in a tube then mixed with 1000 µl of enzyme reagent. • After mixing, determine the cholesterol content in the supernatant with clinical photometer. LDL cholesterol level = total cholesterol – cholesterol in the supernatant

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