SAMPLINC AND ANALYSIS OF COMMERCIAL FATS AND OILS

AOCS:r AOCS Recommended Practice Cd 1b'87

Revised 201 2-

lodine Value of Fats an d o ls

yclo hexane M eth d
DEFINITION
The iodine value is a measure of the unsaturation of fats and oils and is expressed in terms of the number
of centigrams of iodine absorbed per gram of test sample (o/o iodine absorbed).
SCOPE
Applicable to all normal fats and oils'with iodine values in the range of 15 to 70 that do not contain
conjugated double bonds (see Notes, 1).

APPARATUS :I

l.Glass-stoppered iodine flasla-5OO mL.

2. Glass-stoppered volumetric flasls-I000 mL, for preparing standard solutions.
3. dispenser-25 mL, I mL adjustabiliry for lVijs solution.
Volumetric
4. dispenser-25 mL, I mL adjustabiliry, for l5o/o potassium iodide (KI) solution.
Volumetric
5. dispenser-2 mL, 1 mL adjustability, for starch solution.
Volumetric
6. dispenser-50 mL, I mL adjustabiliry for distilled water.
Volumetric
7. Repeater pipet-with filling flask, 20 mL, for cyclohexane.
8. Analytical balance-accurate to 0.0001 g.
9. Magnetic stirrer.
10. Filter paper-\(hatman #541, or equivalent.
I l. Beakers-50 mL.
12. Hot air oven.
13. Timer.

REAGENTS
1. Vijs 5sfulien-466urately standardized (see Notes, 2 and Caution).
2. Potassium iodide (KI) solution-l 5 o/o, prepared by dissolving I 5 g of reagent-grade KI in 100 mL of distilled water.
3. Cyclohexane-reagent grade (see Notes, 3 and4 and Caution).
4. Soluble starch-tested for sensitivity (see Notes, 5). Make a paste with I g of starch (see Notes, 6) and a small amount of
cold distilled water. Add, while stirring, 200 mL of boiling water.
Testfor sensitiuity-Place 5 mL of starch solution in 100 mL of water and add 0.05 mL of freshly prepared|.I M KI solu-
tion and one drop of a 50 ppm chlorine solution made by diluting 1 mL of a commercial 5%o sodium hypochlorite (NaOCl)
solution to 1000 mL. The deep blue color produced must be discharged by 0.05 mL of 0.1 M sodium thiosulfate.
5. Potassium dichromate-reagent grade. The potassium dichromate is finely ground and dried to constant weight at about
110"C before using (see Notes, Caution).
6. Hydrochloric acid-reagent grade, concentrated, sp gr l.l9 (see Notes, Caution).
7. Sodium thiosulfate (NarSrOr.5H2O)-0.1 M, accurately standardized, prepared from reagent-grade NarSrOr.5HrO (see
Notes, T).
Standardization of sodium thiosulfate-\7eidn 0.16-0.22 g of finely ground and dried potassium dichromate into a
500 mL flask or bottle by difference from a weighing bottle. Dissolve in25 mL of water, add 5 mL of concentrated hydro-
chloric acid,20 mL of potassium iodide solution, and rotate to mix. Allow to stand for 5 min, and then add 100 mL of
distilled water. Titrate with sodium thiosulfate solution, shaking continuously until the yellow color hx almost disappeared.
Add l-2 mL of starch indicator and continue the titration, adding the thiosulfte solution slowly until the blue color just
disappears. The strength of the sodium thiosulfate solution is expressed in terms of its molariry.

20.394 x wt of K CrrOr, g
Molarity of NarSrO, solution =
volume, mL of sodium thiosulfate

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SAMPLINC AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 1b-87 . lodineValue of Fats and Oils Cyclohexane Method

PROCEDURE
1. Melt the test sample, if it is not already liquid (the temperature during melting should not exceed the melting point of the
test sample by more than l0"C), and filter through two pieces of filter paper to remove any solid impurities and the last traces
of moisture. The filtration may be performed in an air oven at S0-S5"C, but should be completed within 5 min t 30 sec. The
test sample must be absolutely dry.
Note-NI glassware must be absolutely clean and completely dry.
2. Nter filtration, allow the filtered test portion to achieve a temperature of 68-71 t 1'C before weighing the test porrion.
3. Once the test portion has achieved a temperature of 68-71 t 1'C, immediately weigh the test porrion into a 500 mL iodine
flask, using the weights and weighlng accvtacy noted in Thble I (see Notes, 8).
4. Add 20 mL of ryclohexane on top of the test portion and swirl to ensure that the test portion is completely dissolved (see
Notes, 4).
5. Dispense 25 mL of Vijs solution into flask containing the test portion, stopper the flask and swirl ro ensure an intimare
mixture. Immediately set the timer for 1.0 or 2.0 hr, depending on the iodine value of the test portion: IV < 150, 1.0 hr;
IV > 150, 2.0 hr (see Notes, 4).
6. Store the flasks in the dark at a temperature of 25 t 5'C (see Notes, 9).
7. Prepare and conduct at least one blank determir"ration with each group of test portion simultaneously and similar in all
resPects to the test portion.
8. Remove the flasks from storage and add 20 mL of KI solution, followed by 100 mL of distilled water (see Notes, 10).
9. Titrate with 0.1 M NarSrO, solution, adding it gradually and with constant and vigorous shaking (see Notes, 11). Continue
the titration until the yellow color has almost disappeared. Add l-2 mL of starch indicator solution and continue the
titration until the blue color has just disappeared.

CATCUTATIONS
(B-S) x Mx 12.69
1. The iodine value =
mass, g of test ponion

\7here-
B = volume of titrant, mL of blank
S = volume of titrant, mL of test porrion
M = molariry of NarSrO, solution

PRECISION
l. See Thble 2.

NOTES
Caution
Cyclohexane is flammable and a dangerous fire risk. It is moderately toxic by inhalation and skin conracr. The TLV in air is
300 ppm.
'Wijs
solution causes severe burns, and the vapors can cause lung and eye damage. Use of a fume hood is recommended. \7ijs
solution without carbon tetrachloride is available commercially.
Hydrochloric acid is a strong acid and will cause severe burns. Protective clothing should be worn when working with this
acid. It is toxic by ingestion and inhalation and is a strong irritant to eyes and skin. The use of a properly operaring fume hood is
recommended. tVhen diluting the acid, always add the acid to the wat€r, never the reverse.
Potassium dichromate is toxic by ingestion and inhalation. There is sufficient evidence in humans for the carcinogenicity of
chromium [+6], in particular, lung cancer. It is a strong oxidizing agent and a dangerous fire risk when in conracr with organic
chemicals.

NUMBERED NOTES
l.S(hen the iodine value is determined on materials having conjugated systems, the result is not a measure of total unsatura-
tion, but rather is an empirical value indicative of the amount of unsaturation present. Reproducible resuhs are obtained
which afford a comparison of total unsaturation.
2. Because the preparation of the lVijs solution is time-consuming and involves the use of both hazardous and toxic chemicals,
this solution may be purchased from a chemical supplier. Solutions are available which contain no carbon tetrachloride, and
such solutions should be used. All'S7ijs solutions are sensitive to temper4ture, moisture and light. Store in a cool and dark
place, and never allow to come to a temperature above 25-30"C.
3. Fresh cyclohexane should be used. Erratic results may be obtained ifold cyclohexane is used.
4. The AOCS study reported.in J. Am. Oil Chem.,Sac. (References, l) used ryclohexane alone. 'Vhen using cyclohexane alone,
the 30 min reaction time specified in the former AOCS version of the Vijs method is insufficient for test porrions having an
IV > 100, especially fish oils, as reported in References, 1 . A study completed by FOSFA International in 1992 (References,
2), showed that the ISO 3961 (1989) andAOCS Recommended Practice Cd lb-87 (1993) methods for determining IV do

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 SAMPLINC AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 1b-87 . lodineValue of Fats and Oils Cyclohexane Method

not give rise to values that are statistically different, and that the'use of ryclohexane (alone) in place of carbon tetrachloride
as the solvent does not affect the observed IV for titration of triglyceride oils having iodine values between 18 and 52 (the
range of iodine values encountered in the study). Variable results were obtained in the application of the method to fish oils;
however, further studies carried out by the International Association of Fish Meal Manufacturers indicated that reproducible
IV results could be obtained if the reaction time of the \flijs reagent and fish oil was extended.
5. lolo starch solution may be purchased from a chemical supplier.
6. "Potato Starch for lodometry" is recommended, because this starch produces a deep blue color in the presence of the iodo-
nium 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. The following are suitable starches: Soluble Starch for Iodometry Fisher
5516-100; Soluble Potato Starch, Sigma 5-2630; Soluble Potato Starch for Iodometry, J.T. Baker 4006-04.
7. The sodium thiosulfate solution may be purchased from a chemical supplier. However, it still must be accurately standard-
ized in the laboratory.
8. The weight of the test portion must be such that there will be an excess of \X/ijs solution of 50-600/o of the amount added;
i.e., 100-150o/o of the amount absorbed.
9. If the reaction is not terminated within 3 min past the reaction time, the test portion must be discarded.
10. The test porrion musr be titrated within 3Q min of reaction termination, after which the analysis is invalid.
11. Mechanical stirring is recommended for agitation during the addition of thiosulfate.

REFERENCES
1. AOCS collaborative study results using ryclohexane alone appear inJ. Am. Oil Chem. Soc. 65:745 (1988).
2. INFORM 3:1246 (1992).

Table 1
Test sample weights.
Test sample weight
Iodine value 100o/o excess I 500/o excess \Teighing
^ccr$^qt
o
<3 10 l0 + 0.001
) 10.576 8.461,3 0.005
5 6.346 5.0770 0.0005
l0 3.1730 2.5384 0.0002
20 r.5865 1.2720 0.0002
40 .7935 .6346 0.0002
60 .5288 .4231 0.0002
80 .3966 .3173 0.0001
100 .3t73 .2538 0.0001
120 .2644 .2t15 0.0001
t40 .2266 .1813 0.0001
160 .1983 .1587 0.0001
180 .1762 .t410 0.0001
200 .1586 .1269 0.0001

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Griglt O 2012 by & AOC
 SAMPLINC AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 1b-87 . lodineValue of Fats and Oils Cyclohexane Method

Table 2
The interlaboratory study Gompleted by FOSFA lnternational in 1989, in which 37 laboratories participated, each obtaining
four test results for each test portion analyzed by AOCS Recommended Practice Cd 1 b-87 (cyclohexane) and four test
results for each test portion analyzed by ISO method 3961 (carbon tetrachloride), gave the statistical results (evaluated in
accordance with ISO 5725-19861summarized in the following table.
Test Samples'

Name of statistic P3O-A P3O-B P3O.A P3O-B

No. of laboratories 37 37 37 37
No. of laboratory returns 29 28 35 35
No. of accepted results 29 27 34 33
Mean 52.09 18.17 52.79 18.18
Repeatabiliry standard deviation 0.1I 0.05 0.20 0.08
Repeatability coefficient of variation (o/o) 0.20 0.28 0.39 0.44
Reproducibility standard deviation 0.32 0.14 0.49 0.16
Reproducibiliry coefficient of variation (o/o) , 0.6l 0.77 0.94 0.89
Repeatabiliry (r) 0.30 0.15 0.57 0.22
Reproducibiliry (R) 0.90 0.40 1.39 0.46
Range of accepted results 51.4-52.89 17.8-18.43 50.7-53.3 17.88-18.55
R/r 3.01 ) -71
2.45 2.05

'Test Sample key: P30-A, RBD palm oil; P30-B, crude palm kernel oil.

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