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

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

3. 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
(ml)
H2SO4 (ml)
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

4. * 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