Ethanol Emulsion Test for Fats and Oils

The Ethanol Emulsion Test is a food test which determines the presence of a broad group of
naturally occurring compounds known as lipids. Lipids consist of fats and oils.
Other lipid tests include the Grease Spot Test and the Sudan Stain Test. The Grease spot
test is performed on fats - lipids which are solid at room temperature. Sudan stain colours
lipids red, but is a less common bench reagent than ethanol. The Ethanol Emulsion Test is the
most common test amongst the three.
Procedure
Soild sample
!.Crush the food sample and place in a dry test tube.
".Add ethanol to about " cm# abo$e the le$el of the sample and shake thoroughly.
#.%llow the solid to settle &about # min' to allow the lipid to be e(tracted.
).Decant the ethanol into another test tube.
*.%dd " cm# of deionized water to the second test
tube.
+.,ake obser$ations.
Liquid sample
!.%dd a few drops of the li-uid food sample to a dry test tube.
".%dd " cm# ethanol and shake it thoroughly
#.%dd " cm# of deionized water.
).,ake oser!ations.
"esults and #nterpretation
OBSERVATION INTERPRETATION
POS#T#$E
TEST
% la%er of cloud% white suspensionforms at the top of the
solution. &.pon close inspection you can see the tiny globules
of fat suspended in the solution. This an emulsion. /oods
with high lipid content ha$e a 0higher1 layer than foods with
less'.
Lipids are present
&EGAT#$E
TEST
Solution remains colourless. 2o emulsion is formed. Lipids are not present
Principle of the Ethanol Emulsion Test
The soluilities of lipids and ethanol are e'ploited in this test.
Lipids are non(polar organic compounds. 3ence they are soluble in organic sol$ents such as
ethanol &alcohol', but insoluble in water.
Ethanol is an organic substance and so dissol$es other organic substances4 it is fre-uently
used as an organic sol$ent.
3owe$er ethanol is also miscile in water due to the presence of theh%dro'%l )(
O*+ functional groups and the shortness of its chain &"5'. The hydro(yl group participates
in h%dro,en ondin, with water (see below).
The h%drophoic interaction of the carbon in the short chain with water is not great and is
o$ercome by the hydrogen bonding.
Ethanol e'tracts the lipid from the crushed solid sample. %s ethanol is miscible with lipids no
change is seen upon its addition to the solid and li-uid samples.
The lipid spontaneousl% comes out of solution when water is added and is dispersed as
micelles &small droplets' throughout the solution of ethanol and water.& This happens as
hydrophobic portion of the lipid molecules pro6ect inwards and e(cludes the a-ueous
en$ironment4 the hydrophilic portion &-5OO3' group faces the a-ueous en$ironement.'
% layer is formed at the top as lipids are less dense than water. The droplets diffract li,ht,
appearing cloudy white.
A Conclusi!e Test
% positi!e test shows conclusi!el% that lipids are present - and not the other ma6or
biological molecules
-. Caroh%drates
 "educin, su,ars and non( reducin, su,ars - slightly soluble in ethanol and soluble
in water
 Starch 7 insoluble in both ethanol and water
". Proteins 7 insoluble in ethanol &the addition of ethanol is used to precipitate proteins'
#. &ucleic Acids 7 are insoluble in ethanol and soluble in water
2o change is seen in a ne,ati!e test as there are no lipids to come out of solution.
E'perimental S/ills
The Ethanol Emulsion Test procedure is suitable for assessment of the 0easurement and
0anipulation E(perimental skill.
8elow are some criteria your teacher may use to assess you. 3e will be scrutini9ing the
steps you take to pre$ent contamination , including cross contamination from other food
samples.
C"#TE"#A
:reparation of a Solid Sample
!. 5ompletely crushes ade-uate amount of food with a clean and dry mortar and pestle.
". Transfers an ade-uate amount of crushed food sample to a clean test tube.
#. %dds "cm# ethanol abo$e the food sample.
). Shakes the test tube thoroughly and safely &using a $orte( mi(er if a$ailable'.
*. %llows the solid to settle for about # min.
+. ;ecants the ethanol carefully, into a clean, dry test tube without transference of food
particles.
<. %dds "cm# of de-ioni9ed water to the test tube.
=. 5ompares each result with the negati$e control.
>. 5ompares the heights of the emulsion layer of all the samples.

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