Special Properties of Fats, AACC Method 58-16

Oils, and Shortenings Page 1 of 3

Determination of Peroxide ValueAcetic Acid-Chloroform

Method
First approval October 26, 1977; Reapproval November 3, 1999

Objective
To determine the peroxide value of fats and oils, in terms of meq peroxide per
1000 g of sample. The method is applicable to all normal fats and oils including
margarine. It is highly empirical, and any variation in procedure may result in
variation in results. This method is equivalent to Official Method Cd 8-53 of the
American Oil Chemists Society (AOCS).

Apparatus
1. Pipet, Mohr, measuring type, 1-ml capacity.
2. Erlenmeyer flasks, glass-stoppered, 250-ml.

Reagents
1. Acetic acid-chloroform solution. Caution. See Note 1. Mix 3 parts by
volume glacial acetic acid, reagent grade, with 2 parts by volume chloroform,
U.S. Pharmacopeia grade. See Note 2.
2. KI solution, saturated solution of KI, ACS grade, in recently boiled distilled
water. Make sure solution remains saturated as indicated by presence of undis-
solved crystals. Store in dark. Test daily by adding 2 drops starch solution (see
Note 3) to 0.5 ml KI solution in 30 ml acetic acid-chloroform solution. If blue
color is formed that requires more than 1 drop 0.1N sodium thiosulfate solution
to discharge, discard KI solution and prepare fresh solution.
3. Sodium thiosulfate solution, 0.1N, accurately standardized.
4. Sodium thiosulfate solution, 0.01N, accurately standardized. This solution
may be prepared by accurately pipetting 100 ml 0.1N solution into 1000-ml
volumetric flask and diluting to volume with recently boiled distilled water.
5. Starch indicator solution, 1.0% of soluble starch in distilled water.

Procedure for Fats and Oils

1. Weigh 5.00 0.05 g sample into 250-ml glass-stoppered Erlenmeyer flask
and add 30 ml acetic acid-chloroform solution. Swirl flask until sample is dis-
solved in solution. Add 0.5 ml saturated KI, preferably using Mohr type meas-
uring pipet.
2. Allow solution to stand with occasional shaking for exactly 1 min and then
add 30 ml distilled water. See Note 4.
3. Titrate with 0.1N Na2S2O3, adding gradually and with constant and vigorous
shaking. Continue titration until yellow color has almost disappeared. Add about
0.5 ml starch indicator solution. Continue titration, shaking flask vigorously near
end point to liberate all iodine from chloroform layer. Add thiosulfate dropwise
until blue color has just disappeared. See Note 5.
Special Properties of Fats, AACC Method 58-16
Oils, and Shortenings Page 2 of 3

Determination of Peroxide ValueAcetic Acid-Chloroform

Method (continued)

4. Conduct blank determination of reagents daily. Blank titration must not

exceed 0.1 ml 0.1N Na2S2O3 solution.

Calculation
( S B )( N )( 1000)
Peroxide value as milliequivalents peroxide per 1000 g sample =
weight of sample
where B = titration of blank, S = titration of sample, N = normality of Na2S2O3
solution.

Procedure for Margarine

1. Melt sample by heating with constant stirring on hot plate set at low heat, or
by heating in air oven at 6070. Avoid excessive heating and particularly
prolonged exposure of oil to temperatures above 40.
2. When completely melted, remove sample from hot plate or oven and allow
to settle in warm place until aqueous portion and most of milk solids have settled
to bottom.
3. Decant oil into clean beaker and filter through Whatman no. 4 paper (or
equivalent) into another clean beaker. Do not reheat unless absolutely necessary
for filtration. Sample should be clear and brilliant.
4. Proceed with test on decanted oil as described in steps 14 in Procedure for
Fats and Oils (above).

Notes
1. Chloroform is a known carcinogen. It is toxic by inhalation and has
anesthetic properties. Avoid contact with the skin. Prolonged inhalation or
ingestion can lead to liver and kidney damage and may be fatal. It is nonflamma-
ble but will burn on prolonged exposure to flame or high temperature. The
threshold limit value (TLV) is 10 ppm in air. A fume hood should be used at all
times when using chloroform.
Acetic acid in the pure state is moderately toxic by ingestion and inhalation. It
is a strong irritant to skin and tissue. The TLV in air is 10 ppm.
2. Isooctane has been proposed as a replacement for chloroform in this
method. The method using isooctane is now available as AOCS Official Method
Cd 8b-90. The acetic acid-chloroform version of the method is intended for
deletion by AOCS.
3. Potato starch for iodometry is recommended, because this starch produces
a deep blue color in the presence of the iodonium ion. Soluble starch is not
recommended because a consistent deep blue color may not be developed when
some soluble starches interact with the iodonium ion.
Special Properties of Fats, AACC Method 58-16
Oils, and Shortenings Page 3 of 3

Determination of Peroxide ValueAcetic Acid-Chloroform

Method (continued)

4. The test should be carried out in diffuse daylight or in artificial light

shielded from a direct light source. A report on a coulometric method for the
measurement of peroxide value indicates that the iodide-peroxide reaction is
complete at the end of 1 min and that the liberation of iodine is affected by light.
5. If the titration is less than 0.5 ml using 0.1N sodium thiosulfate, repeat the
determination using 0.01N sodium thiosulfate.

References
1. American Oil Chemists Society. 1998. Official Methods and Recommended Practices, 4th ed.
Method Cc 8-53. The Society, Champaign, IL.
2. AOAC International. 1995. Official Methods of Analysis of AOAC International, 16th ed.
Method 965.33. The Association, Gaithersburg, MD.