TESTS METHODOLOGY

Trans fat (TFA) analysis

Using FTIR Spectroscopy (AOAC and AOCS methods)

Melting point
Open-tube Capillary-Slip Method
The temperature at which the oil or fat softens or becomes sufficiently fluid to slip or run as
determined by the open-tube capillary-slip method
Ref: AOCS Official Method Cc 3-25
Time: 30 minuts

Free fatty acids

By simple titration method with KOH
Time: 30 minuts

Refractive index
Measurement of the refractive index of the sample is done by means of a suitable refractometer.
Ref :- A.O.A.C 17th edn, 2000, Official method 921.08 – Index of refraction of oils and fats
Time: 15 mins
Specific gravity
Melt sample if necessary. Filter through a filter paper to remove any impurities and the last
traces of moisture. Make sure that the sample is completely dry. Cool the sample to 300C or
ambient temperature desired for determination
By using Pycnometer
Ref :- A.O.A.C 17th edn, 2000, Official method 920.212 Specific gravity (Apparent) of Oils,
Pycnometer method

Peroxide value
The fat or oil is dissolved in a chloroform-acetic acid mixture and subjected to an excess of
iodide via a saturated solution of potassium iodide. The peroxides present oxidize the iodide to
iodine and the iodine is then titrated to a colorimetric endpoint using sodium thiosulfate with
starch as an indicator. The amount of iodine produced is directly proportional to the peroxide
value.
Ref: A.O.A.C. 17th edn, 2000,Official Method 965.33 Peroxide Value in Oils and Fats
Time: 1hr

Acid value
The acid value is determined by directly titrating the oil/fat in an alcoholic medium against
standard potassium hydroxide/sodium hydroxide solution
IUPAC 2.201(1979) / I.S: 548 (Part 1) – 1964, Methods of Sampling and Test for Oils and Fats

Sponification Number
The oil sample is saponified by refluxing with a known excess of alcoholic potassium hydroxide
solution. The alkali required for saponification is determined by titration of the excess potassium
hydroxide with standard hydrochloric acid.
Ref:- A.O.A.C 17th edn, 2000, Official method 920.160 Saponification number of oils and fats
Time : 1 hour

Iodine value
The oil/fat sample taken in carbon-tetrachloride is treated with a known excess of iodine
monochloride solution in glacial acetic acid. The excess of iodine monochloride is treated with
potassium iodide and the liberated iodine estimated by titration with sodium thiosulfate solution.
Ref: AOCS Official Method Cd 1b-87 : Iodine value of fats and oils

P-anisidine value by spectrophotometer (Aldehydes+ketones)

Aldehydes, derived from the secondary oxidation of fat matrix, react with the p-anisidine
determining a variation in the absorbance, measured at 366 nm. Anisidine value is expressed as
AnV (Anisidine value) following AOCS (Cd 18-90) the reference method. Anisidine value test
is used to assess the secondary oxidation of oil or fat, which is mainly imputable to aldehydes
and ketones, and is therefore able to tell the oxidation “history” of an oil or a fat.
Furthermore, AnV analysis on oil is an indicator of excessive oil deterioration in deep frying
process.

Total polar compounds (TPCs/TPM)

By oil testometer
Time: 5 min

Color
By absorption (Spectrophotometer)
Total oxidation value
PoV + P-AnV