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Estimation of Total Lipids by Colorimetric Method. * Principle. LIPIDS. H 2 SO 4. 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.
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Estimation of Total Lipids by Colorimetric Method * Principle LIPIDS 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
60 5 55 10 50 45 15 40 20 35 25 30 15min. 10 min. St. Unk. St. Unk. 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 Absorbance of Unknown Absorbance of Standard = Concentration of Unknown Concentration of Standard CUn. = A Un. × C St. A St. Samar A. Damiati