AV =

ml of KOH x N x 56
Weight of Sample
= mg of KOH
ANALYTICAL METHODS TO MEASURE THE CONSTANTS OF FATS
AND OILS
[1] Acid value (Acid Numbe!
The acid value (AV) is the number that expresses, in milligrams the quantity of potassium
hydroxide required to neutralize the free acids present in 1 g of the substance. The acid value
may be overestimated if other acid components are present in the system, e.g. amino acids or
acid phosphates. The acid value is often a good measure of the break down of the triacylglycrols
into free fatty acids, which has an adverse effect on the quality of many lipids.
Si"#i$ica#ce
cid value is the measure of hydrolytic rancidity. !n general, it gives an indication about edibility
of the lipid.
" #dible oil contain $ 1%
" &harmaceutical oil must not have any acidity.
' ( 'ormality of )*+
% ,ree ,atty cid (,,- ( . x /.0/1
Ma%eial
,at or *il
bsolute ethanol alcohol
&henolphthalein
/.1 ' )*+
&'cedue
1. &lace 0./ g of fat or oil in a dried conical flask.
2. dd 20 ml of absolute ethanol alcohol and add ( 2"1- drops of phenolphthalein
1. +eat with shaking in water bath (30%- for 1/ minutes ,then cool Titrate the solution
against /.1 ' )*+ until pink color appears (end point-.
4. 5ecord your observations.
0. 6alculate the acid value (.- and free fatty acid (%,,- using above laws.
728 Sa('#i$ica%i'# Numbe

*
6 5
*
*
6 5
6 5
*
+
2
6 *
+6 *
+
2
6 *
)*+
+
+
+
+
2
6 *
+6 *
+
2
6 *
5 6 *)
+ 3
+
3
The saponification value is the number of mg of potassium hydroxide required to neutralize
the free acids and to saponify the esters in 1 g of the substance. The saponification number is a
measure of the average molecular weight of the triacylglycerols in a sample. 9aponification is
the process of breaking down a neutral fat into glycerol and fatty acids by treatment with alkali.
The smaller the saponification number the larger the average molecular weight of the
triacylglycerols present i.e. 9aponification value is inversely proportional to the mean molecular
weight of fatty acids (or chain length-.
9aponification .alue of ,ats and *ils
,at or oil 9aponification .alue
:ilk fat 21/"211
6oconut oil 20/"234
6otton seed oil 1;<"1<;
9oybean oil 1;<"1<0
lard 1</"2/2
=utter fat and vegetable fats 22/ > 20/
:aterial
,at or *il
/.0 ' alcoholic potassium hydroxide ( alcoholic )*+- ( prepared by dissolving 1/ g
potassium hydroxide in 2/ m? of water and make the final volume to 1 ? using <0 %
ethanol. ?eave the solution to stand for 24 h before decanting and filtering the solution.
/.0 ' +ydrochloric acid
&henolphthalein.
&rocedure
1. @eigh approximately 2 g of the fat or oil into a 20/ m? conical flask.
2. dd 20 m? of alcoholic potassium hydroxide solution ( /.0 '-.
1. ttach a reflux condenser and heat the flask contents on a boiling water bath for 1 hour
with occasional shaking.
SP# =
56.1! "S# x N of H$l
%&am of Sample
4. @hile the solution is still hot , add 1 drops of phenolphthalein indicator and titrate the
excess potassium hydroxide with the /.0 ' hydrochloric acid ( .ml of hydrochloric acid
at end point represents 9-.
0. Ao same above procedure but without sample ( .ml of hydrochloric acid at end point
represents =-.
3. calculate the saponification number by using the following lawB

718 #ster .alue
T)e e*%e value is defined as the mg of )*+ required to react with glycerin (glycerol C or
glycerin- after saponify one gram of fat. !t is calculated from the saponification
.alue (SV) and the acid .alue (.-B

E*%e +alue (E+! , Sa('#i$ica%i'# +alue (S+! - Acid +alue (A+!
. glycerin = Ester Value /0/12332
748 !odine .alue (!..-
The iodine value (IV- gives a measure of the average degree of unsaturation of a lipidB the
higher the iodine value, the greater the number of 6(6 double bonds. =y definition %)e i'di#e
value i* e4(e**ed a* %)e "am* '$ i'di#e ab*'bed (e 1//" '$ li(id0 !odine value (!...- is
directly proportional to the degree of unsaturation ('o of double bonds.- and inversely
proportional to the melting point (:.&.- of lipid. n increase in !...indicates high susceptibility
of lipid to oxidative rancidity due to high degree of unsaturation.
*ne of the most commonly used methods for determining the iodine value of lipids is
D+anus methodD. The lipid to be analyzed is weighed and dissolved in a suitable organic solvent,
to which a known excess of iodine chloride is added. 9ome of the !=r reacts with the double
bonds in the unsaturated lipids, while the rest remainsB
5"6+(6+"5 E !=rexcess 5"6+!"6+=r"5 E !=rremaining
The amount of !=r that has reacted is determined by measuring the amount of !=r remaining
after the reaction has gone to completion (!=rreacted (!=rexcess " !=rremaining-. The amount of !=r



=B ml of +6l required by =lank.

9B ml of +6l required by 9ample.
remaining is determined by adding excess potassium iodide to the solution to liberate iodine, and
then titrating with a sodium thiosulfate ('a292*1- solution in the presence of starch to determine
the concentration of iodine releasedB
!=rremaining E 2)! )=r E )! E !2
!2 E starch E 2'a292*1 (blue- 2'a! E starch E 'a294*3 (colorless-
TY&ICAL IODINE NUM5ERS
6oconut oil ; " 1/
=utter 20 " 4/
=eef tallow 1/ " 40
&alm oil 1F " 04
?ard 40 " F/
*live oil F0 " <0
&eanut oil ;0 " 1//
6ottonseed oil 1// " 11F
6orn oil 110 " 11/
,ish oils 12/ " 1;/
9oybean oil 120 " 14/
9afflower oil 11/ " 14/
9unflower oil 11/ " 140
?inseed oil 1F/ " 2/0
:aterial
*il or fat
+anus solution ( itGs prepared by dissolving 1;.2 g of iodine in 1? of glacial acetic acid
and then add 1 ml of bromine water for increasing the halogen content.
10% potassium iodide solution
1% starch solution
/.1 ' 9odium thiosulfate solution.
&rocedure
1. @eigh approximately /.20 g of the fat or oil into a 20/ m? conical flask.
2. dd 1/ ml of chloroform.
1. dd 1/ ml of +anus solution and close the flask completely by &ara film, then
leave the solution for 1/ minutes with shaking continuously.
4. dd 1/ ml of 10% potassium iodide solution and then shake.
0. dd 1// ml of distilled water (A@-.
3. Titrate the iodine solution against /.1 ' 9odium thiosulfate solution till yellow
color formed , then add 2"1 drops of starch solution where blue solution formed
and then continue with titration till the blue color is disappeared (.olume (ml- of
'a292*1 at end point represents 9-
F. Ao same above procedure but without sample (.olume (ml- of 'a292*1 at end
point represents =-.
;. 6alculate the iodine number by using the following lawB
( = " 9 - ' of 'a292*1 /.12FgCmeq 1//
!odine .alue (
@eight of 9ample (g-
=B . ml of 'a292*1 volume for blank
9B . ml of 'a292*1 volume for sample