Acidity AACC International Method 02-02.

02
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Fat Acidity—Rapid Method, for Small Grains

Final approval October 3, 1984; Reapproval November 3, 1999; Revision 2009

Objective
Natural fats are mixtures of esters of fatty acids with glycerol. They are essen-
tially neutral. Unfavorable storage conditions may cause partial hydrolysis of
the glycerides. The resultant free fatty acids increase the acidity, which is an
indication of deterioration in quality. This method determines total fatty acids in
small grains by a rapid titrimetric procedure.

Apparatus
Grinder-extractor.

Reagents
1. Toluene, purified grade.
2. Alcohol-phenolphthalein solution (0.04%). To 1 liter ethyl alcohol (95%),
add 0.4 g phenolphthalein.
3. KOH. Prepare CO2-free standard solution 0.0356N (1 ml = 2 mg KOH).
4. Color standards. The intensity of yellow color in grain varies, depending on
type of grain; therefore, a color standard is helpful in making titration end points
uniform. Prepare as follows:
To 50 ml water in flask of type used for titrating, add dropwise 0.05% potas-
sium dichromate until water solution matches in color the grain extract solution
to be titrated. Add 2.5 ml freshly prepared 0.01% potassium permanganate solu-
tion and mix. Color of titration end point should match this standard. Prepare
color standard for titration blank by adding 2.5 ml 0.01% potassium perman-
ganate to 50 ml water.

Procedure
For most accurate results, moisture content of grain should not exceed 11.0%.
For small grains such as wheat, weigh 40 g of representative sample and transfer
to cup of mill. See Note.
1. Mix ground material to establish uniformity, weigh 40 g, and transfer to
cup of Stein mill.
2. If sample was previously ground, add 100 ml toluene to 40 g of ground
material in cup. If sample was not previously ground, attach cup to mill and run
mill 1 min before adding 100 ml toluene.
3. After adding toluene, attach cup to mill and run mill 4 min. Remove cup
from mill and cover with watch-glass until ready to filter.
4. Filter sample as quickly as possible, preferably with vacuum, through
funnel fitted with Whatman no. 2 filter paper. If vacuum is not available, filter
through folded filter paper. Cover funnel with watch-glass while filtering.
5. Transfer 25-ml aliquot of filtrate to titration flask (125-ml Erlenmeyer).
Add 25 ml alcohol-phenolphthalein solution.

doi: 10.1094/AACCIntMethod-02-02.02
Acidity AACC International Method 02-02.02
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Fat Acidity—Rapid Method, for Small Grains (continued)

6. Titrate grain solution with 0.0356N KOH. End point should match color of
standard described under Reagents.
7. Determine blank titration by adding 25 ml alcohol-phenolphthalein solution
to 25 ml toluene and titrating to end point matching color of standard for
titration blank described under Reagents.
Calculation
Report fat acidity as mg KOH required to neutralize free fatty acids from 100
g grain on dry-matter basis by formula:
(T − B ) × ( N ) × (56.10) × (100)
Fat acidity value = × 100
Wt × (100 − M )
where T = volume of sample titration (ml), B = volume of blank titration (ml),
N = normality of titrant (mmol/ml), Wt = weight of sample (g), and M = mois-
ture of sample (%).

Note
For larger grains such as corn, first grind in laboratory mill about 200 g of
representative sample.

References
1. Baker, D. 1984. The use of toluene in fat acidity methods. Cereal Foods World 29:265.
2. Baker, D., Neustadt, M. H., and Zeleny, L. 1957. Application of the fat acidity test as an index of
grain deterioration. Cereal Chem. 34:226.
3. Zeleny, L., and Coleman, D. A. 1938. Acidity in cereals and cereal products, its determination and
significance. Cereal Chem. 15:580.
4. Zeleny, L., and Coleman, D. A. 1939. The chemical determination of soundness in corn. U.S.
Dep. Agric., Tech. Bull. 644.