METTLER TOLEDO Application M624-2012

Determination of Peroxide Value in Edible Oils and Fats

The sample is dissolved in a solvent mixture of acetic acid and chloroform and then with a solution of
potassium iodide. The liberated iodine is titrated with standard solution of sodium thiosulfate.

Sample Olive oil, Sunflower oil, Preparation and Procedures

Groundnut oil, Palm oil,
CAUTION:
Coconut oil, Margarine (3- 5 g)
Work in a fume hood and use safety goggles, wear
Compound Peroxide functional group gloves and a lab coat.
R-OOH
Preparation
Chemicals 20 mL acetic acid:Chloroform 3:2 0.01 mol/L sodium thiosulfate:
1 mL saturated KI solution - Weigh 2.48 g sodium thiosulfate in a 1000 mL
50 mL deionized water volumetric flask.
- Stir to dissolve and then fill up to the mark with
Titrant Sodium thiosulfate, Na2S2O3
deionized water and shake well before
c(Na2S2O3) = 0.01mol/L
transferring to a bottle.
Saturated KI solution:
- Add 127 g potassium iodide in 100 mL volumetric
Standard Potassium iodate, KIO3 flask. Stir to dissolve with 100 mL with deionized
M= 213.99 g/mol , z = 6 water.

Preparation: standard solution Procedures

in 0.01 M in 0.1 HCl • Blank determination
- Add 20 mL diluent solution (acetic acid:
Indication DMi140-SC
Combined platinum ring electrode chloroform 3:2) into the beaker.
- Add 1mL sat. KI solution and stir for 180 s
- Add 50 mL deionized water followed by
→
- +
Chemistry ROOH + 2I + 2H3O titration with with 0.01 mol/L Na2S2O3.
ROH + I2 + 3H2O
• Sample determination
I2 + 2 Na2S2O3 → - Accurately weigh 3- 5g of sample into the
titration beaker and add 20 mL diluent
2 NaI + Na2S4O6
solution (acetic acid: chloroform 3:2), and stir
for 30 s.
- Add 1 mL saturated KI solution and stir for
180 s
Calculation R1 =
(Q - B[Blank Peroxide value])*C/m - Add 50 mL deionized water followed by
C = 1000, meq/kg titrate with sodium thiosulfate.

B[Blank Peroxide Value] = Remarks

Blank value
Q = Titrant consumption (mmol) - The method parameters have been optimized
m = Sample size in g for the above samples. It may be necessary to
slightly adapt this method for your specific
samples.
Waste Halogenated organic waste - Beakers have to be cleaned very well. Invisible
disposal fat residues or traces of metal can affect the
result strongly
Author, Vineesh Pallath, IMSG, May 2012
Rev: Cosimo De Caro, - Literature: Swiss Food Manual,
Version
MSG AnaChem, June 2012 SLMB (“Schweizerisches Lebensmittebuch”).

METTLER TOLEDO Page 1 of 6 Titration Application M624-2012

 Instruments - T70/T90 Titration Excellence
(With minor method modification also a T50 Titration Excellence can be used).
- Additional dosing unit (ME-51109030)
- DV1010 burette 10 mL (ME-51107501), DV1020 burette 20 mL (MT-51107502)
®
Accessories - LabX pro titration software
- XP205 Balance
- Rainin 1mL pipette

Results
Peroxide value of Groundnut oil.

Blank: Method ID: BlankPeroxide2

Results

No. Comment/ID Start time sample size Results

1/3 Blank 5/17/2012 1:50:39PM 20mL R1= 0.001 mmol consumption
2/3 Blank 5/17/2012 1:59:40PM 20mL R1= 0.001 mmol consumption
3/3 Blank 5/17/2012 2:11:21PM 20mL R1= 0.001 mmol consumption
R2= 0.0010 mmol Mean Blank

Blank: Statistics

Rx Name n Mean value Unit s srel[%]

R1 Consumption 3 0.001 mmol 0.000 0
R2 Mean Blank EQP 1 0.0010 mmol NaN NaN

Sample: Method ID: Peroxidevalue2

No Comment/ID Start time Sample size Results

1/6 Groundnut Oil 5/17/2012 2:25:35PM 4.53470 g R1= 0.0577 mmol Consumption
R2= 12.507 meq/kg Content
2/6 Groundnut Oil 5/17/2012 2:39:43PM 4.67944 g R1= 0.0593 mmol Consumption
R2= 12.468 meq/kg Content
3/6 Groundnut Oil 5/17/2012 2:49:53PM 4.53069 g R1= 0.0577 mmol Consumption
R2= 12.512 meq/kg Content
4/6 Groundnut Oil 5/17/2012 3:09:05PM 4.72777 g R1= 0.0603 mmol Consumption
R2= 12.552 meq/kg Content
5/6 Groundnut Oil 5/17/2012 3:20:57PM 4.51762 g R1= 0.0574 mmol Consumption
R2= 12.493 meq/kg Content
6/6 Groundnut Oil 5/17/2012 3:34:37PM 4.76492 g R1= 0.0605 mmol Consumption
R2= 12.481 meq/kg Content

Sample: Statistics

RX Name n MeanValue Unit s srel[%]

R1 Consumption 6 0.0588 mmol 0.0014 2.376
R2 Content 6 12.502 meq/kg 0.029 0.235

Peroxide Values of different oils and fats

Sample Sample size (g) n Mean Value Unit s srel[%]

Olive Oil 3.0 - 5.0 6 10.830 meq/kg 0.116 1.067

Sunflower Oil 3.0 - 5.0 6 2.995 meq/kg 0.089 2.971
Palm Oil 3.0 - 5.0 6 3.585 meq/kg 0.074 2.059
Margarine 3.0 - 5.0 6 1.177 meq/kg 0.057 4.815
Coconut Oil 3.0 - 5.0 6 0.239 meq/kg 0.01 4.324

METTLER TOLEDO Page 2 of 6 Titration Application M624-2012

 Titration curve

Blank value determination of Groundnut Oil Peroxide Value determination of Groundnut Oil

Table of measured values

Blank value determination of Groundnut Oil.
Volume Increment Signal Change 1st deriv. Time Temperature
mL mL mV mV mV/mL s °C
0.0000 NaN 270.3 NaN NaN 0 25.0
0.0100 0.0100 270.8 0.5 NaN 3 25.0
0.0200 0.0100 269.8 -1.0 NaN 6 25.0
0.0450 0.0250 264.7 -5.1 NaN 10 25.0
0.0640 0.0190 259.0 -5.7 NaN 13 25.0
0.0825 0.0185 149.0 -10.0 -931.99 16 25.0
0.0925 0.0100 235.9 -13.1 -717.51 22 25.0
EQP1 0.095141 NaN 229.8 NaN -954.16 NaN NaN
0.1025 0.0100 212.7 -23.2 -565.61 33 25.0
0.1125 0.0100 205.3 -7.4 -625.00 40 25.0
0.1375 0.0250 195.7 -9.6 -518.85 48 25.0
0.1770 0.0395 188.1 -7.6 NaN 55 25.0
0.2590 0.0820 179.7 -8.4 NaN 62 25.0
0.3590 0.1000 174.1 -5.6 NaN 67 25.0
0.4590 0.1000 171.2 -2.9 NaN 71 25.0
0.5590 0.1000 169.5 -1.7 NaN 74 25.0

Peroxide value determination of Groundnut Oil.

Volume Increment Signal Change 1st deriv. Time Temperature
mL mL mV mV mV/mL s °C
0.000 NaN 316.4 NaN NaN 0 25.0
0.0100 0.0100 316.9 0.5 NaN 3 25.0
0.0200 0.0100 317.2 0.3 NaN 6 25.0
0.0450 0.0250 317.4 0.2 NaN 9 25.0
0.1075 0.0625 317.5 0.1 NaN 12 25.0
0.2075 0.1000 317.6 0.0 1.10 15 25.0
--- --- --- --- --- --- ---
5.2075 0.1000 294.8 -1.9 -18.56 168 25.0
5.3075 0.1000 292.7 -2.1 -19.59 170 25.0
5.4075 0.1000 290.2 -2.5 -23.28 174 25.0
5.5075 0.1000 287.0 -3.2 -37.21 177 25.0
5.6075 0.1000 282.7 -4.3 -69.16 180 25.0
5.7075 0.1000 276.1 -6.6 -164.05 183 25.0
5.8075 0.1000 263.1 -13.0 -444.41 187 25.0
5.8460 0.0385 249.6 -13.5 -667.09 190 25.0
5.8560 0.0100 243.1 -6.5 -499.13 193 25.0
EQP1 5.865024 NaN 229.9 NaN -555.27 NaN NaN
5.8660 0.0100 228.5 -14.6 -551.76 202 25.0
5.8760 0.0100 205.3 -23.2 -542.60 215 25.0
5.8860 0.0100 196.2 -9.1 -600.58 224 25.0
5.9085 0.0225 187.6 -8.6 -569.80 232 25.0
5.9585 0.0500 177.4 -10.2 -313.61 240 25.0
6.0320 0.0735 168.5 -8.9 NaN 248 25.0
6.1320 0.1000 163.6 -4.9 NaN 253 25.0

METTLER TOLEDO Page 3 of 6 Titration Application M624-2012

 Comments

General remarks:
- Fats and oils undergo changes during storage due to oxidation with air (oxidative rancidity). The
compounds formed react as peroxides. Their content is commonly expressed as the "peroxide value"
in milliequivalents peroxidic oxygen per kg of sample. Peroxide values higher than 10 – 20 meq
O2/kg are commonly interpreted as rancidity.
- Unrefined oils show peroxide values. They are usually in the range between 5 – 20 meq O2/kg.
Peroxide values are expressed either in milliequivalent/kg (meq/kg) or millimoles/L (mmol/L).
- The duration of the generation time (time between adding KI solution, adding of deionized water and
the starting of titration) is the critical point for repeatable results. As an example, the influence of stir
time has been studied and found that 180 s is the optimal stirring time:

Stir time (s) Result (meq/kg)

60 11.563

180 10.830

600 10.656

- Based on the sample solubility, different solvents can be used such as acetic acid: chloroform (3:2),
acetic acid: cyclohexane (3:2) and acetic acid: isooctane (3:2).
- The sample has to be protected from the air, stored in a cool, place and should not be opened before
the determination is commenced. The samples will undergo oxidation with air (oxidative rancidity).
Olive oil was analyzed after exposing to air and sunlight, the peroxide value found to be changed
from 10.830 meq/kg to 14.640 meq/kg, hence support this statement.
- Automation can be achieved with a Rondo/Rondolino sample changer (setting conditioning with
deionized water for 30-60 seconds).
- Use 0.1M sodium thiosulfate titrant when the expected peroxide value is above 20 meq/kg and use
0.01M sodium thiosulfate titrant when the expected peroxide value is less than 20 meq/kg.
- Clean the electrode thoroughly with plenty of deionized water each and before every titration.
Procedures:
1. Titer determination.
2. Blank value determination.
Impurities of the solvent are taken into account by a blank value determination.
The blank value determination is done under the same conditions as the peroxide value
determination itself.
The blank value is automatically stored as B[Blank Peroxide value] in the instrument setup.
3. Sample titration.

Reaction Scheme:
- Generation of hydroperoxides:
R-H + O2 → R-OOH (Reaction I)
- Generation of iodine:
→
- +
KI + CH3COOH HI + CH3COO K (Reaction II)
ROOH + 2HI → ROH + H2O + I2 (Reaction III)
- Titration step:
I2 + 2Na2S2O3 → Na2S4O6 + 2 NaI (Reaction IV)

METTLER TOLEDO Page 4 of 6 Titration Application M624-2012

 Method
Add. EQP criteria No
Blank value determination Termination
At Vmax 5.0 mL
001 Title At potential No
Type General titration At slope No
Compatible with T70/T90 After number of
ID BlankPeroxide2 recognized EQPs Yes
Title Peroxide value Blank 180S Number of EQPs 1
Author admin Combined termination
Date/Time 5/14/2012 11:39:13 AM criteria No
Modified at 5/14/2012 11:39:13 AM Condition
Modified by admin Condition No
Protect No
SOP None 010 Calculation R1
Result Consumption
002 Sample Result unit mmol
Number of IDs 1 Formula R1=Q
ID 1 Blank Constant C= 1
Entry type Fixed volume M M[None]
Fixed volume 20 mL z z[None]
Density 1.0 g/mL Decimal places 3
Correction factor 1.0 Result limits No
Temperature 25.0°C Record Statistics Yes
Extra statistical
003 Titration stand (Manual stand) functions No
Type Manual stand Send to buffer No
Titration stand Manual stand1 Condition No

004 Dispense (normal) [1] 011 End of sample

Titrant AA/Chloroform
Concentration 100 012 Blank
Volume 20 mL Name Blank Peroxide Value
Dosing rate 60.0 mL/min Value B= Mean[R1]
Condition No Units mmol
Limits No
005 Stir Condition No
Speed 40%
Duration 130 s 013 Calculation R2
Condition No Result Mean Blank EQP
Result unit mmol
006 Instruction Formula R2=Mean[R1]
Instruction Add 1 mL KI Constant C= 1
Mode Confirmation M M[None]
Print Yes z z[None]
LabX command No Decimal places 4
Condition No Result limits No
Record statistics Yes
007 Stir Send to buffer No
Speed 30% Condition No
Duration 180s
Condition No

008 Instruction
Instruction Add Water
Mode Confirmation
Print Yes
LabX command No
Condition No

009 Titration (EQP) [1]

Titrant
Titrant Na2S2O3
Concentration 0.01 mol/L
Sensor
Type mV
Sensor DM140-SC
Unit mV
Temperature acquisition
Temperature acquisition No
Stir
Speed 30%
Predispense
Mode None
Wait time 0 s
Control
Control User
Titrant addition Dynamic
dE(set value) 8.0 mV
dV(min) 0.01 mL
dV(max) 0.1 mL
Mode Equilibrium controlled
dE 0.5 mV
dt 1.0 s
t(min) 3.0 s
t(max) 130.0 s
Evaluation and recognition
Procedure Standard
Threshold 200 mV/mL
Tendency None
Ranges 0

METTLER TOLEDO Page 5 of 6 Titration Application M624-2012

 Termination
Sample titration At Vmax 10.0 mL
At potential No
001 Title At slope No
Type General titration After number of
Compatible with T70/T90 recognized EQPs yes
ID PeroxideValue2 Combined termination
Title Peroxide value 180S Number of EQPs 1
Author admin Combined termination
Date/Time 5/14/2012 11:45:55 AM criteria No
Modified at 5/14/2012 11:45:55 AM Accompanying stating
Modified by admin Accompanying stating No
Protect No Condition
SOP None Condition No

002 Sample 010 Calculation R2

Number of IDs 1 Result Consumption
ID 1 Olive Oil Result Unit mmol
Entry type Weight Formula R1=Q
Lower limit 0.0 [g] Constant C= 1
Upper limit 5.0 [g] M M[None]
Correction factor 1.0 z z[None]
Temperature 25.0°C Decimal places 4
Entry Arbitrary Result limits No
Record statistics Yes
003 Titration stand (Manual stand) Extra statistical
Type Manual stand Functions No
Titration stand Manual stand1 Send to buffer No
Condition No
004 Dispense (normal) [1]
Titrant AA/Chloroform 011 Calculation R2
Concentration 100 Result Content
Volume 20 mL Result unit meq/kg
Dosing rate 60.0 mL/min Formula R2=(Q-B[Blank Peroxide
Condition No value])*C/m
Constant C= 1000
005 Stir M M[None]
Speed 40% z z[None]
Duration 30 s Decimal places 3
Condition No Result limits No
Record Statistics Yes
006 Instruction Extra statistical
Instruction Add 1mL KI Functions No
Mode Confirmation Send to buffer No
Print Yes Condition No
LabX command No
Condition No 012 End of sample

007 Stir
Speed 30%
Duration 180s
Condition No

008 Instruction
Instruction Add water
Mode Confirmation
Print Yes
LabX command No
Condition No

009 Titration (EQP) [1]

Titrant
Titrant Na2S2O3
Concentration 0.01 mol/L
Sensor
Type mV
Sensor DM140-SC
Unit mV
Temperature acquisition
Temperature acquisition No
Stir
Speed 30%
Predispense
Mode None
Wait time 0 s
Control
Control User
Titrant addition Dynamic
dE(set value) 8.0 mV
dV(min) 0.01 mL
dV(max) 0.1
Mode Equilibrium controlled
dE 0.5 mV
dt 1.0
t(min) 3.0
t(max) 30.0
Evaluation and recognition
Procedure Standard
Threshold 200 mV/mL
Tendency None
Ranges 0
Add. EQP criteria No

METTLER TOLEDO Page 6 of 6 Titration Application M624-2012