Fat and Oil Peroxide value of Oleic acid

Redox titration by
Automatic Potentiometric Titrator
Test Method for Analysis of Basic
Fatty Material
Standard
ISO 3960

1. Abstract
The oleic acid is widely distributed in plant and animal fats. Measurement of
peroxide value (POV) of oleic acid is demonstrated in this application according to the
below quoted test method. The test sample is first dissolved in mixture of chloroform
and acetic acid23. By flowing nitrogen gas through the sample to dispel residual
oxygen, add potassium iodide, and then titrate free iodine with 0.01mol/L sodium
thiosulfate.
The endpoint is determined by the maximum inflexion point on titration curve.
POV is calculated from titration volume of sodium thiosulfate.

I2 + 2Na2S2O3 Na2S4O6 + 2NaI

2. Reference
1) Test Method for Analysis of Basic Fatty Material by Japanese Oil Chemistry
Society; Reference material 2.4-1996 Peroxide value Chloroform Method
2) ISO 3960 : 2007 Animal and vegetable fats and oils Determination of peroxide
value

3. Cautions in measurement
1) Do not use water containing carbon dioxide in preparing saturated potassium
iodide solution.
2) To confirm saturation, add excessive potassium iodide to the saturated
potassium iodide solution, and leave crystal in the solution.
3) Prepare fresh saturated potassium iodide each time.
4) If the sample does not change color to yellow due to free iodine when saturated
potassium iodide is added, add more saturated potassium iodide except when
blank test is performed or if peroxide value is very small in number.
5) Before potassium iodide is added, flow nitrogen gas through the sample to dispel
residual oxygen.
6) If potential change around EP is small, adjust Gain of Control parameter.

TIB-99309 Ver.01 1/4

4. Post-measurement care
Rinse the electrode with pure water, and keep it dipped in water to avoid drying up.

5. Test equipment
Main unit Automatic potentiometric titrator Standard preamplifier: STD
Electrode Option Combination Pt. electrode

6. Reagent
Reagent 0.01mol/L Sodium thiosulfate 1.006
Solvent Mixture of chloroform and acetic acid (23)
Additive Saturated potassium iodide
Inactive gas Nitrogen gas

7. Measurement procedure
measurement
1) Deliver approx. 5g sample into a conical flask with stopper.
2) Add 30mL solvent and gently shake to dissolve the sample completely.
3) Gently replace the air inside flask with nitrogen to remove remaining oxygen.
4) By further flowing nitrogen gas, add 0.5mL saturated potassium iodide, and
immediately seal the flask and gently shake it for one minute.
5) Leave the flask at room temperature 15 to 20C in a dark room.
6) Add 30mL pure water, and seal it and stir.
7) Titrate with 0.01mol/L sodium thiosulfate to measure peroxide value.
Likewise, obtain blank level in advance by a blank test.

8. Formula
Peroxide value ( meq / kg ) = ( EP1 BL1 ) TF R / SIZE
EP1 Titration volume ( mL )
BL1 Blank level ( 0.00mL )
TF Factor of reagent ( 1.006 )
R Constant ( 10 )
SIZE Sample size ()

TIB-99309 Ver.01 2/4

9. Example of measurement
Ambient condition
Room temperature22.5 Humidity 47 Weather Cloudy

-Titration parameter- -Titration curve-

Model : AT-510 *** R e s u l t ***
Method No. : 11
Titr.mode : Auto Sample No. : 41-01
Intermit Date : 1999/07/06 11:49
Titr.form : EP Stop Sample ID :

[Result parameter] Method No. : 11

[Titration parameter] <Auto Intermit>
<Calculation>
Form : EP Stop Calc. Type : Sample Titr. Time : 00:10:33
APB No. : 1 Conc.1 : Set
Unit No. : 1 CO1=(EP1-BL1)*TF*R/SIZ Size : 5.0794g
Detector No. : 1 E
Unit : mV Conc-1 : 2.6095meq/kg
Max.Volume : 10.0mL Unit : meq/kg
Wait Time : 0s EP No. : 1 End point-1
Direction : Auto Temp.Comp. : Off Volume : 1.3176mL
Potential : 213.3mV
<Constant>
[Control parameter] R : 10

End Point No. : 1 <Titr. Constant>

End sense : Auto Factor : 1.006
End Point Area : Off Conc : 0.01
Separation : Off
Over Titr.Vol. : 0mL <Blank>
Gain : 10 Blank1 : 0.00
Data samp.Pot. : 4.0mV
Data samp.Vol. : 0.1mL
Stability : 0.5mV/s
Delay Time : 1s
Limit Time : 0s

(The above printout data were obtained from titration by AT-510 unit)
Titration parameter
Form: of titration / APB No. the burette used in titration / Unit No.: APB Unit File number
Detector No.: the detector used in titration / Max Volume: of titration / Wait Time: before titration starts
Direction: of titration
Control parameter
End Point No. number of EPs detected / End sense: direction of EP / End Point Area: detection area
Separation: of potential / Over Titr.Vol. over-titration volume / Gain: sensitivity of detection signal
Data samp.Pot.: potential changes of sampling signal / Data samp.Vol.: titration volume of sampling signal
Stability: of titration / Delay Time: before stability check / Limit Time: for stability check
Result parameter
Calc.Type: of formula / Conc.1 formula 1 / Unit of result
EP No. for calculation / Temp.Comp. temperature compensation of titration liquid / R: constant
Factor: of reagent / Blank1: blank level 1

TIB-99309 Ver.01 3/4

Measurement results
Sample Titration POV
n Peroxide value
g mL meq / kg
1 5.0794 1.3176 2.6095 Mean 2.6267 meq/kg
2 5.0043 1.3053 2.6241 SD 0.0186 meq/kg
3 5.0095 1.3179 2.6465 RSD 0.7095 %
The above results were obtained by 3 tests of the same sample.
Red underline shows the data from page 3/4.

10. Summary
The oleic acid reduces bad cholesterolLDLof monounsaturated fatty acids but does
not reduce good cholesterolHDL. Recently, such advantage attracts public
attention, and cooking oil like canola oil or olive oil which contains oleic acid is on
the market. Unsaturated fatty acid like oleic acid and linoleic acid are liable to be
oxidized with ambient oxygen. Its quality can be controlled by measuring POV.
The measurement results this time shows favorable repeatability with 0.7% relative
standard deviation. Precise and reliable measurement is assured by the automated
potentiometry.
POV of oleic acid can be perfectly measured by any of the following titration systems
manufactured by Kyoto Electronics (KEM).

AT-610 AT-510 AT-500N-1

Awarded Product of Supreme Compact and cost performance Low cost and high performance
Technology from Kyoto City model Easy view with back light LCD
Easy key entry by touch panel of PC card expands data memory for GLP/GMP conformed model
large color LCD 8-inch wide convenience and versatility.
Simultaneous titration in parallel
Both potentiometric and Karl
Fischer moisture titration
(coulometricvolumetric can be
performed at a time.

TIB-99309 Ver.01 4/4