BIOCHEMISTRY – LAB Term01

BSMT MED201

QUALITATIVE TEST FOR LIPIDS

SOLUBILITY TEST
 Preliminary test that
detects the presence of
lipids
- Alcohol (ethanol) –
(mixed but considered
as polar)
- Water - polar
- Chloroform - nonpolar EMULSIFICATION TEST
 Detects lipid solubility in various solvents to  Used to detect the
check whether it is miscible or immiscible in presence of lipids
polar or non-polar solvents
 Process that stabilizes the
 Lipids are nonpolar molecules, which means water and oil emulsion by
their ends are not charged. Because they are using the emulsifying
nonpolar and water is polar, lipids are not agents
soluble in water.
 Emulsifying agents (bile salts/soap) emulsify
 Positive in chloroform and ethanol (lipids can the lipid → tiny droplets suspended in the
dissolve) solution

TRANSLUCENT SPOT TEST SAPONIFICATION

 Preliminary test for the  Tests for Ester
lipids
 Determines the presence
 Translucent and greasy of an ester functional
spot group in a solution
- Due to the greasiness
 Saponifiable or non-
of lipids
saponifiable
 Alkaline hydrolysis of the fatty acid esters
TEST FOR UNSATURATION (Bromine Test)
 Bromine can be used to test for the presence of
multiple bonds
 Bromine → brownish color
 When added → Alkene or Alkyne (multiple
bonds) the
 When added → Alkane (single bond) brownish End product: Glycerol/soap
color remains
BROMINE TEST
 QUALITATIVE TESTS FOR LIPIDS - BIOCHEMLAB

SUDAN IV TEST LIEBERMANN-BURCHARD TEST

 Detect presence of lipids  Cholesterol reacts with strong conc.
 Principle: binding and acid (sulphuric acid & acetic
solubility of lipid in non-polar anhydride)
compunds - Sulphuric acid: Dehydrading
 Sudan IV is a non-polar agent
stain that readily stains - Acetic anhydride: Oxidizing agent
lipids red-orange  Acetic Anhydride test
 Used for the detection of cholesterol
ACROLEIN TEST  Formation of a green or green-blue color after
 Used to detect glycerol and a few minutes is positive
fat
 Based on dehydration Liebermann-Burchard Test
reaction in which water
molecules are removed
from glycerol by adding
potassium hydrogen sulphate.
 When a fat is heated strongly in the presence of
a dehydrating agent such as KHSO4, the glycerol
portion of the molecule is dehydrated to form Polymer containing a chromophore (Green)
the unsaturated aldehyde (acrolein)
 Pungent smell and the black color indicate the
presence of glycerol

SUDAN IV TEST AND ACROLEIN TEST Prepared by:

Cherries V. Sanchez, RMT

DICHROMATE TEST
 Used to detect the
presence of glycerol
 Based on the principle
Oxidation reaction →
Glycerol and Dichromate
ions react → Brown color
solution (-)
 Chromic ions oxidize the glycerol in the
presence of nitric acid→ blue color solution (+)
(chronous ions)