+ + Estimation of Total Lipids by Colorimetric Method * Principle H2SO4 Carbonium ions + + Vanillin phosphate ester Purple Complex

Materials Phosphovanillin reagent Sulfuric acid Standard solution of cholesterol (1g/100 ml acetone) Standard solution of olive oil

Mix Well on a vortex mixer Mix Well Read the Optical Density at 540 nm 15min. Cholesterl Olive oil Blank 10 min. St. Unk. St. Unk. 60 30 15 45 5 10 20 25 35 40 50 55 20µ unk. 20µ unk. 20µ st. 20µ st. 20µ D.W. Phosphovanillin 10 10 10 10 10 (ml) H2SO4 (ml) 0.2 0.2 0.2 0.2 0.2 Mix Well on a vortex mixer Mix Well COOL in water at Room Temperature Incubate at 37ºC in water bath Read the Optical Density at 540 nm

* Calculations CUn. = A Un. × C St. Absorbance of Unknown Absorbance of Standard = Concentration of Unknown Concentration of Standard CUn. = A Un. × C St. A St. Samar A. Damiati