1 / 9

Estimation of Cholesterol Lieberman-Burchard Reaction

Cholesterol is a lipidic, waxy alcohol found in the cell membranes and transported in the blood plasma of all animals. Cholesterol is readily soluble in acetone, while most complex lipids are insoluble in this solvent. . Estimation of Cholesterol Lieberman-Burchard Reaction .

salim
Download Presentation

Estimation of Cholesterol Lieberman-Burchard Reaction

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Cholesterol is a lipidic, waxy alcohol found in the cell membranes and transported in the blood plasma of all animals. Cholesterol is readily soluble in acetone, while most complex lipids are insoluble in this solvent. Estimation of Cholesterol Lieberman-Burchard Reaction

  2. Estimation of Cholesterol Liberman-Burchard Reaction • Blood or serum is extracted with an alcohol-acetone mixture which removes cholesterol and other lipids and precipitates protein.

  3. Estimation of Cholesterol Liberman-Burchard Reaction • Liberman-Burchard Reaction: Acetic anhydride reacts with cholesterol in a chloroform solution to produce a characteristic blue-green color. • Liberman-Burchard Test: A test for unsaturated steroids

  4. Estimation of Cholesterol Liberman-Burchard Reaction • When conc. sulfuric acid is added to cholesterol, a water molecule is removed from C3 of cholesterol molecule, and it is oxidised to form 3,5-cholestadiene.This product is converted to a polymer containing a chromophore which results in the green colouration indicating a positive test.

  5. Estimation of Cholesterol Liberman-Burchard Reaction - H2O + H2SO4 3,5-cholestadiene polymer containing a chromophore (blue-green color)

  6. Unk. Shaking the mixture 5 min Boiling Water Bath until the solvent begins to boil COOL to Room Temperature 0.2ml blood 10ml alcohol-aceton solvent Centrifuge tube

  7. Read the Optical Density at 680 nm 2ml Acetic anhydride-sulfuric acid mixture 2ml Chloroform Leave the tubes in the dark at room remperature

  8. Tube # Blank St. Unk. Acetic anhydride-sulfuric 2 2 2 acid mixture (ml) Leave the tubes in the dark at room remperature Chloroform (ml) 2 2 2 Read the Optical Density at 680 nm D.W (ml) 0.2 0 0 Unknown (ml) 0 0 0.2 Mix Well Standard (ml) 0 0.2 0

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

More Related