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Column chromatography is another solidliquid technique in which the two phases are
a solid (stationary phase) and a liquid
(moving phase). The theory of column
chromatography is analogous to that of thinlayer chromatography. The most common
adsorbents - silica gel and alumina - are the
same ones used in TLC. The sample is
dissolved in a small quantity of solvent (the
eluent) and applied to the top of the column.
The eluent, instead of rising by capillary
action up a thin layer, flows down through
the column filled with the adsorbent. Just as
in TLC, there is an equilibrium established
between the solute adsorbed on the silica gel
or alumina and the eluting solvent flowing
down through the column.
Stationary phase (adsorbent)
The stationary phase or adsorbent in
column chromatography is a solid. The most
common stationary phase for column
chromatography is silica gel, followed by
alumina. Cellulose powder has often been
used in the past. Also possible are ion
exchange chromatography,reversed- phase
chromatography
(RP),
affinity
chromatography or expanded bed adsorption
(EBA). The stationary phases are usually
finely ground powders or gels and/or are
microporous for an increased surface,
though in EBA a fluidized bed is used.
65:25:4
(v/v/v)
ether:methanol:NH4OH.
petroleum
The
Lieberman-Burchard
or
acetic
anhydride test is used for the detection of
cholesterol. The formation of a green or
green-blue color after a few minutes is
positive.
Lieberman-Burchard is a reagent used in a
colorimetric test to detect cholesterol, which
gives a deep green color. This color begins
as a purplish, pink color and progresses
through to a light green then very dark green
color. The color is due to the hydroxyl group
(-OH) of cholesterol reacting with the
reagents and increasing the conjugation of
the un-saturation in the adjacent fused ring.
Based on the results that were culled (Table
2), the first and third eluate did not produce
any change in color. The second eluate
REFERENCES:
BOOKS:
Column Chromatography
http://orgchem.colorado.edu/hndbksupport/c
olchrom/colchrom.html
Bettelheim,F.A.,
March,J.
(1990).
Introduction to organic and biochemistry.
Philadelphia: Saunders College.
Lipid Library
Heftman, E. (1967). Chromatography. New
York: Reinhold Publishing Corporation
http://lipidlibrary.aocs.org/Lipids/whatlip/in
dex.htm#def
Retrieved: March 8, 2010
Lehninger,
A.L.
(2008).
Legninger
Principles of Biochemistry. New York: W.H.
Freeman.
McKee.
(2003).
Biochemistry:
The
Molecular Basis of Life. Boston: McGrawHill.
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