ESTIMATION OF LIPIDS

Principal:

Total lipids in serum, after hot treatment of Serum with sulphuric acid, react with the phosphoric acid, giving a pink
color, which should be read at wavelength of 530nm.

Apparatus:
Micropipette, colorimeter, cuvette, water bath

Procedure:
1. Carbonization:
 Take 2 test tubes. In one test tube add 2.5ml of H2SO4along with 40ul of standard solution. It is
“Liposulphuric standard”
 In the other test tube, take 2.5ml of H2SO4 along with 40ul of sample. It is “Liposulphuric Sample”
 Stir and incubate at 100C for 10 mins.
 Cool rapidly, Liposulphuric mixture is then obtained.
2. Calorimetric reaction:

1. Switch on the Colorimeter

2. Take 3 cuvettes and label them blank, standard and test.
3. Add 1ml of reagent in all the three cuvettes.
4. Add 40 ul of H2SO4 in the blank one.
5. Add 40 ul of standard Liposulphuric mixture in the standard one.
6. Add 40ul of sample Liposulphuric to sample cuvette.
7. Mix carefully and incubate at room temperature for 15-20 mins.
8.Measure the absorbance of sample and standard against reagent blank with in 45 mins.
9. Measure concentration of Lipids in the Sample using the formula

Calculation:

Conc of lipids in sample = Optical density of test x Conc of stand

Optical density of standard
Conc of standard = 800mg/dl
 Normal range of lipids is 450-800mg/dl

Clinical significance:

Hyperlipidemia/hyperlipoproteinemia/dyslipidemia: It is the presence

of raised or abnormal levels of lipids and/or lipoproteins in the blood. Lipids are t
ransported in a protein capsule, and the density of the lipids and the type of proteins determine the fate of the
particle and its influence on metabolism.