Scribd Logo
Upload

Welcome to Scribd!

  • 271 views

    Estimation of Cholesterol Lieberman-Burchard Reaction: - Cholesterol Is A Lipidic

    Uploaded by

    Reffada Yodhyasena
    The document describes the Liberman-Burchard reaction for estimating cholesterol levels. Blood or serum is extracted with alcohol-acetone to remove cholesterol and other lipids. In the Liberman-Burchard reaction, acetic anhydride reacts with cholesterol in chloroform to produce a characteristic blue-green color. When conc. sulfuric acid is added, it oxidizes cholesterol to form a compound that polymerizes and produces the blue-green color, indicating the presence of cholesterol.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd

    Estimation of Cholesterol Lieberman-Burchard Reaction: - Cholesterol Is A Lipidic

    Uploaded by

    Reffada Yodhyasena
    271 views9 pages
    The document describes the Liberman-Burchard reaction for estimating cholesterol levels. Blood or serum is extracted with alcohol-acetone to remove cholesterol and other lipids. In the Liberman-Burchard reaction, acetic anhydride reacts with cholesterol in chloroform to produce a characteristic blue-green color. When conc. sulfuric acid is added, it oxidizes cholesterol to form a compound that polymerizes and produces the blue-green color, indicating the presence of cholesterol.

    Original Title

    1598638.ppt

    Copyright

    © © All Rights Reserved

    Available Formats

    PPT, PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    The document describes the Liberman-Burchard reaction for estimating cholesterol levels. Blood or serum is extracted with alcohol-acetone to remove cholesterol and other lipids. In the Liberman-Burchard reaction, acetic anhydride reacts with cholesterol in chloroform to produce a characteristic blue-green color. When conc. sulfuric acid is added, it oxidizes cholesterol to form a compound that polymerizes and produces the blue-green color, indicating the presence of cholesterol.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    271 views9 pages

    Estimation of Cholesterol Lieberman-Burchard Reaction: - Cholesterol Is A Lipidic

    Uploaded by

    Reffada Yodhyasena
    The document describes the Liberman-Burchard reaction for estimating cholesterol levels. Blood or serum is extracted with alcohol-acetone to remove cholesterol and other lipids. In the Liberman-Burchard reaction, acetic anhydride reacts with cholesterol in chloroform to produce a characteristic blue-green color. When conc. sulfuric acid is added, it oxidizes cholesterol to form a compound that polymerizes and produces the blue-green color, indicating the presence of cholesterol.

    Copyright:

    © All Rights Reserved
    Download as PPT, PDF, TXT or read online from Scribd
    Download as ppt, pdf, or txt
    of 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 Liberman-
    Burchard Reaction
    • Blood or serum is
    extracted with an
    alcohol-acetone
    mixture which
    removes cholesterol
    and other lipids and
    precipitates protein.
    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
    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.
    Estimation of Cholesterol Liberman-
    Burchard Reaction

    - H2O
    + H SO
    2 4

    3,5-cholestadiene

    polymer containing a chromophore


    (blue-green color)
    Shaking the mixture 5 min
    Unk.
    Boiling Water Bath
    until the solvent begins
    to boil
    COOL to Room Temperature
    0.2ml blood

    10ml alcohol-aceton solvent

    Centrifuge tube
    Read the Optical Density at 680 nm

    2ml Acetic anhydride-sulfuric


    acid mixture
    2ml Chloroform

    Leave the tubes in the dark at room remperature


    Tube # Blank St. Unk.

    Acetic anhydride-sulfuric 2 2 2
    acid mixture (ml)

    Chloroform (ml) 2 2 2
    D.W (ml) 0.2 0 0
    Unknown (ml) 0 0 0.2

    Standard (ml)
    Mix Well
    0 0.2 0

    Leave the tubes in the dark at room remperature


    Read the Optical Density at 680 nm
    * Calculations
    Absorbance of Unknown Absorbance of Standard
    =
    Concentration of Unknown Concentration of Standard

    CUn. = A Un. × C St.


    A St.

    Samar A. Damiati

    You might also like