胆固醇含量测定.ppt

Experiment  DETERMINATION OF SERUM TOTAL- AND HDL-CHOLETEROL  principle Tube H: 0.2ml serum + 0.2ml precipitant Tube SH: 0.4ml standard cholesterol+0.4ml precipitant Stand for 15 min 2000rpm.10min supernatant unit ml Tube NO. T ST H SH B Supernatant -- -- 0.2 0.2 -- Serum 0.02 -- -- -- -- Cholesterol standard -- 0.1 -- -- -- Distilled water 0.18 0.1 -- -- 0.2 Enzyme reagent 2.0 2.0 2.0 2.0 2.0 Calculation AT 0.1 Total cholesterol (mg/dl)= × × 50 AST 0.02 AH HDL-cholesterol (mg/dl)= × 50 ASH * LDL VLDL HDL magnesium phosphotungstate (磷钨酸镁) LDL VLDL Serum cholesterol free cholesterol cholesterol ester RO 以血浆脂蛋白形式运输 HDL保留在上清中 cholesterol esterase Cholesterol ester + H2O free cholesterol + fatty acid cholesterol oxidase Free cholesterol + O2 ? 4-cholestenone + H2O2 2H2O2 + 4-aminoantipyrine + phenol red compound peroxidase +4H2O Enzyme reagent: 4-aminoantipyrine 4-氨基安替比林 phenol, peroxidase Procedure: Mix.Incubate at 37℃.20min, Read the absorbance at 520nm ,B as zero Standard cholesterol(50mg/dl) *