SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS

AOCS Recommended Practice Cd 1b-87

Reapproved 1997

Iodine Value of Fats and Oils

Cyclohexane Method
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 sample (% 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 7. Sodium thiosulfate (Na2S2O3 · 5H2O)—0.1 N, accu-

1. Glass-stoppered iodine flasks—500 mL. rat e ly standard i ze d, prep a red from re age n t - gra d e
2. G l a s s - s t o p p e red vo l u m e t ric flasks—1000 mL, fo r Na2S2O3 · 5H2O (see Notes, 7).
preparing standard solutions. S t a n d a rd i z ation of sodium thiosulfat e — We i g h
3. Volumetric dispenser—25 mL, 1-mL adjustability, for 0.16–0.22 g of finely ground and dried potassium dichro-
Wijs solution. mate into a 500-mL flask or bottle by difference from a
4. Volumetric dispenser—25 mL, 1-mL adjustability, for weighing bottle. Dissolve in 25 mL of water, add 5 mL
15% potassium iodide (KI) solution. of concentrated hydrochloric acid, 20 mL of potassium
5. Volumetric dispenser—2 mL, 1-mL adjustability, for iodide solution and rotate to mix. Allow to stand for 5
starch solution. min, and then add 100 mL of distilled water. Titrate with
6. Volumetric dispenser—50 mL, 1-mL adjustability, for sodium thiosulfate solution, shaking continuously until
distilled water. the yellow color has almost disappeared. Add 1–2 mL of
7. Repeater pipet—with filling flask, 20 mL, for cyclo- starch indicator and continue the titration, adding the
hexane. thiosulfate solution slowly until the blue color just disap-
8. Analytical balance—accurate to 0.0001 g. pears. The strength of the sodium thiosulfate solution is
9. Magnetic stirrer. expressed in terms of its normality.
10. Filter paper—Whatman no. 41H, or equivalent.
11. Beakers—50 mL. Normality of Na2S2O3 solution =
12. Hot air oven. 20.394 × wt of K2Cr2O7, g
13. Timer.
volume, mL of sodium thiosulfate
REAGENTS
1. Wijs solution—accurately standardized (see Notes, 2 and
Caution). Table 1
2. Potassium iodide (KI) solution—15%, prep a red by Sample weights.
dissolving 15 g of re age n t - grade KI in 100 mL of
distilled water. Sample weight
3. Cyclohexane—reagent grade (see Notes, 3 and 4 and Iodine 100% 150% Weighing
Caution). value excess excess accuracy
4. Soluble starch—tested for sensitivity (see Notes, 5).
g g g
Make a paste with 1 g of starch (see Notes, 6) and a
<3 10 10 ± 0.001
small amount of cold distilled water. Add, while stirring,
3 10.576 8.4613 0.005
200 mL of boiling water.
5 6.346 5.0770 0.0005
Test for sensitivity—Place 5 mL of starch solution in
10 3.1730 2.5384 0.0002
100 mL of water and add 0.05 mL of freshly prepared
20 1.5865 1.2720 0.0002
0.1 N KI solution and one drop of a 50 ppm chlorine
40 .7935 .6346 0.0002
solution made by diluting 1 mL of a commercial 5%
60 .5288 .4231 0.0002
sodium hypochlorite (NaOCl) solution to 1000 mL. The
80 .3966 .3173 0.0001
deep blue color produced must be discharged by 0.05 mL
100 .3173 .2538 0.0001
of 0.1 N sodium thiosulfate.
120 .2644 .2115 0.0001
5. Potassium dichromate—reagent grade. The potassium
140 .2266 .1813 0.0001
dichromate is finely ground and dried to constant weight
160 .1983 .1587 0.0001
at about 110°C before using (see Notes, Caution).
180 .1762 .1410 0.0001
6. Hydrochloric acid—reagent grade, concentrated, sp gr
200 .1586 .1269 0.0001
1.19 (see Notes, Caution).

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SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 1b-87 • Iodine Value of Fats and Oils Cyclohexane Method

PROCEDURE almost disappeared. Add 1–2 mL of starch indicator

1. Melt the sample, if it is not already liquid (the tempera- solution and continue the titration until the blue color
ture during melting should not exceed the melting point has just disappeared.
of the sample by more than 10°C), and filter through
two pieces of filter paper to remove any solid impurities CALCULATIONS
and the last traces of moisture. The filtration may be (B − S) × N × 12.69
performed in an air oven at 80–85°C, but should be 1. The iodine value =
completed within 5 min ± 30 sec. The sample must be mass, g of sample
absolutely dry. Where—
Note—All glassware must be absolutely clean and B = volume of titrant, mL of blank
completely dry. S = volume of titrant, mL of sample
2. After filtration, allow the filtered sample to achieve a N = normality of Na2S2O3 solution
t e m p e rat u re of 68–71 ± 1°C befo re weighing the
sample. PRECISION
3. Once the sample has achieved a temperature of 68–71 ± 1. See Table 2.
1°C, immediately weigh the sample into a 500-mL
iodine flask, using the weights and weighing accuracy NOTES
noted in Table 1 (see Notes, 8). Caution
4. Add 20 mL of cyclohexane on top of the sample and Cyclohexane is flammable and a dangerous fire risk. It is
swirl to ensure that the sample is completely dissolved moderately toxic by inhalation and skin contact. The TLV
(see Notes, 4). in air is 300 ppm.
5. Dispense 25 mL of Wijs solution into flask containing Wijs solution causes severe burns, and the vapors can
the sample, stopper the flask and swirl to ensure an inti- cause lung and eye damage. Use of a fume hood is recom-
mate mixture. Immediately set the timer for 1.0 or 2.0 mended. Wijs solution without carbon tetrachloride is avail-
hr, depending on the iodine value of the sample: IV able commercially.
< 150, 1.0 hr; IV ≥ 150, 2.0 hr (see Notes, 4). H y d ro ch l o ric acid is a strong acid and will cause
6. Store the flasks in the dark at a temperature of 25 ± 5°C severe burns. Protective clothing should be worn when
(see Notes, 9). working with this acid. It is toxic by ingestion and inhala-
7. Prepare and conduct at least one blank determination tion and is a strong irritant to eyes and skin. The use of a
with each group of samples simultaneously and similar p ro p e rly operating fume hood is re c o m m e n d e d. Wh e n
in all respects to the sample. diluting the acid, always add the acid to the water, never the
8. Remove the flasks from storage and add 20 mL of KI reverse.
solution, followed by 100 mL of distilled water (see Potassium dichromate is toxic by ingestion and inhala-
Notes, 10). tion. There is sufficient evidence in humans for the carcino-
9. Titrate with 0.1 N Na2S2O3 solution, adding it gradu- genicity of chromium [+6], in particular, lung cancer. It is a
ally and with constant and vigorous shaking (see Notes, strong oxidizing agent and a dangerous fire risk when in
11). Continue the titration until the yellow color has contact with organic chemicals.

Table 2
The interlaboratory study completed by FOSFA International in 1989, in which 37 laboratories participated, each obtaining
four test results for each sample analyzed by AOCS Recommended Practice Cd 1b-87 (cyclohexane) and four test results for
each sample analyzed by ISO method 3961 (carbon tetrachloride), gave the statistical results (evaluated in accordance with
ISO 5725-1986) summarized in the following table.
Samplesa
Name of statistic P30-A P30-B P30-A P30-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
Repeatability standard deviation 0.11 0.05 0.20 0.08
Repeatability coefficient of variation (%) 0.20 0.28 0.39 0.44
Reproducibility standard deviation 0.32 0.14 0.49 0.16
Reproducibility coefficient of variation (%) 0.61 0.77 0.94 0.89
Repeatability (r) 0.30 0.15 0.57 0.22
Reproducibility (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 2.72 2.45 2.05
aSample key: P30-A, RBD palm oil; P30-B, crude palm kernel oil.

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 SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 1b-87 • Iodine Value of Fats and Oils Cyclohexane Method

NUMBERED NOTES 52 (the ra n ge of iodine values encountered in the

1. When the iodine value is determined on mat e ri a l s study). Variable results were obtained in the application
having conjugated systems, the result is not a measure of the method to fish oils; however, further studies
of total unsaturation, but rather is an empirical value carried out by the International Association of Fish
i n d i c at ive of the amount of unsat u ration pre s e n t . Meal Manu fa c t u re rs indicated that rep ro d u c i ble IV
R ep ro d u c i ble results are obtained wh i ch affo rd a results could be obtained if the reaction time of the Wijs
c o m p a rison of total unsat u ration. When the iodine reagent and fish oil was extended.
value is required on fatty acids, the preparation and 5. 1% starch solution may be purchased from a chemical
separation are performed as directed in AOCS Official supplier.
Method Cd 6-38. 6. “ Po t ato Starch for Iodometry” is re c o m m e n d e d,
2. Because the preparation of the Wijs solution is time- because this starch produces a deep blue color in the
consuming and involves the use of both hazardous and presence of the iodonium ion. “Soluble Starch” is not
toxic chemicals, this solution may be purchased from a recommended because a consistent deep blue color may
chemical supplier. Solutions are ava i l able wh i ch not be developed when some soluble starches interact
contain no carbon tetra ch l o ri d e, and such solutions with the iodonium ion. The fo l l owing are suitabl e
should be used. All Wijs solutions are sensitive to starches: Soluble Starch for Iodometry, Fisher S516-
temperature, moisture and light. Store in a cool and 100; Soluble Potato Starch, Sigma S-2630; Soluble
dark place, and never allow to come to a temperature Potato Starch for Iodometry, J.T. Baker 4006-04.
above 25–30°C. 7. The sodium thiosulfate solution may be purchased from
3. Fresh cyclohexane should be used. Erratic results may a chemical supplier. However, it still must be accurately
be obtained if old cyclohexane is used. standardized in the laboratory.
4. The AOCS study reported in J. Am. Oil Chem. Soc. 8. The weight of the sample must be such that there will
(References, 1) used cyclohexane alone. When using be an excess of Wijs solution of 50–60% of the amount
cyclohexane alone, the 30 min reaction time specified added; i.e., 100–150% of the amount absorbed.
in the former AOCS version of the Wijs method is 9. If the reaction is not terminated within 3 min past the
insufficient for samples having an IV >100, especially reaction time, the sample must be discarded.
fish oils, as rep o rted in Refe rences, 1. A study 10. The sample must be titrated within 30 min of reaction
completed by FOSFA Intern ational in 1992 termination, after which the analysis is invalid.
(References, 2), showed that the ISO 3961 (1989) and 11. M e chanical stirring is recommended for agi t at i o n
AOCS Recommended Practice Cd 1b-87 (1993) meth- during the addition of thiosulfate.
ods for determining IV do not give rise to values that
are statistically different, and that the use of cy cl o- REFERENCES
hexane (alone) in place of carbon tetrachloride as the 1. AOCS collaborative study results using cyclohexane
solvent does not affect the observed IV for titration of alone appear in J. Am. Oil Chem. Soc. 65:745 (1988).
triglyceride oils having iodine values between 18 and 2. INFORM 3:1246 (1992).

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