SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS

AOCS Official Method Cd 12b-92

Oil Stability Index

DEFINITION
All oils and fats have a resistance to oxidation which depends on the degree of saturation, natural or
added antioxidants, prooxidants or prior abuse. Oxidation is slow until this resistance is overcome, at
which point oxidation accelerates and becomes very rapid. The length of time before this rapid accel-
eration of oxidation is the measure of the resistance to oxidation and is commonly referred to as the
“induction period.”
In this method for determining the induction period, a stream of purified air is passed through a
sample of oil or fat which is held in a thermostated bath. The effluent air from the oil or fat sample is
then bubbled through a vessel containing deionized water. The conductivity of the water is continual-
ly monitored. The effluent air contains volatile organic acids, swept from the oxidizing oil, that
increase the conductivity of the water as oxidation proceeds. Formic acid is the predominant organic
acid formed (see References, 1). The conductivity of the water is monitored by a computer or strip
chart recorder. The Oil Stability Index (OSI) is defined as the point of maximum change of the rate
of oxidation, or mathematically as the maximum of the second derivative of the conductivity with
respect to time (see Fig. 1). This time-based end point may be determined by a computer that can cal-
culate the maximum of the second derivative with respect to time, or by a slope-change algorithm,
which is similar to detecting the onset of peaks for integration of GLC chromatograms. The end point
may be approximated by using other methods. One commonly used approximation is a graphic
method in which tangents are drawn manually (see Fig. 2). The OSI may be run at temperatures of
100, 110, 120, 130 and 140°C. Because by its nature this analysis has this temperature flexibility, all
OSI results should specify the OSI time, with the analysis temperature reported immediately after
(for example, “OSI 11.7 hours at 110°C”).
SCOPE
This method is applicable in general to all fats and oils and has been subjected to a collaborative
study (see References, 2) covering a broad range of sample types. It can be used to analyze crude
oils, or other oils that are prone to foaming, if one drop of silicone antifoam is added prior to analy-
sis. It may be used for other types of oils outside the range of samples tested in the collaborative
study (see Notes, 1 and References, 3). This analysis is an automated replacement for the Active
Oxygen Method (AOM) for fat stability, AOCS Official Method Cd 12-57.

GENERAL PRECAUTIONS remain adhered to a glass surface even after the most
1. Trace-metal contamination of the glassware will cause thorough rinsing with deionized water. Water used for
accelerated oxidation. Because of the difficulty in rinsing should be checked as a potential source of
removing the final traces of chromate from a glass sur- trace-metal contamination.
face, chromate cleaning solutions should not be used. 2. Improper temperature control is the most likely source
Only detergents without surface-active agents should of error. The temperature must be calibrated by check-
be used for cleaning. Traces of surface-active agents ing the actual temperature of a sample in the bath. The

Raw data

Intersection of
2nd Derivative
tangents

1st Derivative

Time Time
Figure 1. OSI determined by first and second derivatives. Figure 2. OSI determined by tangential method.
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 SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 12b-92 • Oil Stability Index

temperature must be maintained within at least tube. An all-glass assembly, either Martin M 7920, or
± 0.1°C. The temperature of the water in the effluent Brinkmann EA1087, or equivalent, is satisfactory.
trap must not exceed 25°C, which minimizes the loss 8. Conductivity measurement tube and probe—The tube
of formic acid. should be capable of holding 50 mL water such that a
depth of 5 cm or more covers the tip of the aeration
APPARATUS tube to provide contact time with the water. This is
1. Two commercial instruments embody the equipment necessary for the volatile organic acids to dissolve. The
needed for this analysis: conductivity probe should supply a full-scale sensitivi-
(a) Oxidative stability instrument—from Omnion Inc., ty of 300–500 uS-cm−1.
Maple and Plain Streets, Rockland, MA, 02370, 9. Instrumentation for detecting conductivity and means for
USA (manufactured under license from Archer recording and evaluating the end point—A computer or
Daniels Midland Co., Decatur, IL, USA); and multichannel strip chart recorder may be used to monitor
(b) Rancimat—model 617 from Brinkmann Instru- the conductivity of each probe in the instrument.
ments, Inc., subsidiary of Sybron Corporation, 10. Tube-cleaning bath—consisting of a stainless steel pan
Cantiague Road, Westbury, NY, 11590, USA. or sink, heated with either steam coils (note: do not use
2. Constant temperature bath—to maintain all samples at copper tubing) or gas, and of sufficient area to permit
the desired temperature to within ± 0.1°C. laying the required number of aeration and test tubes
3. Air distributing manifold—constructed of stainless on the bottom.
steel, plastic, aluminum or glass. Airflow between 11. Test tube brush—fan tip, nylon.
sample channels should be calibrated to permit the 12. Tongs—stainless steel or nickel plated, suitable for
same airflow to within 10%. This may be accom- handling tubes in hot detergent solution.
plished by using either matched capillary tubes or nee- 13. Hood.
dle valves and flow meters.
4. Source of low pressure, clean, oil-free, low CO2, com- REAGENTS
pressed dry air (see Notes, 2)—5.5 pounds per square 1. Water—deionized or distilled (conductivity < 5 µS-cm−1).
inch (PSI) is sufficient to supply air to 24 sample tubes. 2. 1 , 1 , 1 - Trichloroethane, purified grade (see Notes,
The air may be purified with the system as follows: Caution).
(a) Air inlet tube from compressed air source 3. Acetone—ACS Grade (see Notes, Caution).
equipped with stainless steel needle valve or pres- 4. Detergent—for cleaning glassware without leaving
sure regulator. contaminating residue. Contrad 70, Liquinox, Citranox
(b) A Balston Model 75-45 Purge Gas Dryer for FTIR RBS (Fluka Ltd., Buchs, Switzerland) and Micro are
and a Balston A963-000 Trace Hydrocarbon satisfactory.
remover. Balston, Inc., Naperville, IL 60563, or its 5. Silicone antifoam—Dow Corning A. One drop is
equivalent. added to oils that have a tendency to foam.
(c) Bottled air may be used, provided that it has low
CO2 levels (<2 ppm CO2). CLEANING OF SAMPLE REACTION TUBES
5. Thermometer—accurately calibrated, able to indicate The following procedures are either for instruments that do
the temperature to within ± 0.1°C. For OSI operation, a not utilize disposable glassware, or for precautionary clean-
precision thermometer calibrated against an NIST ing of disposable glassware.
traceable thermometer is required. Brooklyn 1. Initial cleaning—The initial cleaning should be per-
Thermometer, Farmingdale, NY provides such a ther- formed immediately after the preceding determination.
mometer with a temperature range of 95–150°C in Melt and drain off as much of the fat from sample tube
0.1°C finest readable interval, 76 mm immersion. The as possible. Wash off the remaining fat with a suitable
stock # is 3551 RM. Brooklyn Thermometer will pro- solvent. 1,1,1-Trichloroethane is a satisfactory fat and
vide NIST traceable calibration for an additional oil solvent.
charge. For temperature calibration of the Rancimat 2. Tube cleaning—Prepare a 1% solution of detergent
Analyzer, an NIST traceable calibrated platinum resis- (Reagents, 4) and heat almost to boiling in the cleaning
tance (RTD) digital thermometer is required with cali- bath. Rinse each tube with the hot detergent solution,
bration points at 100, 110, 120, 130 and 140°C. Omega brushing briefly with the nylon brush (Apparatus, 11).
Engineering, Inc., Stanford, CT, provides such a ther- Then place the tubes in the hot detergent solution con-
mometer. A custom 3 wire 2 mm × 7 mm sensor probe tained in the cleaning bath (Apparatus, 10) in such a
is required for calibration of sample temperature with manner that no air bubbles are trapped within. Boil
air flowing through the sample. vigorously for 30 min. Brush each tube vigorously
6. Gas flow meter—either a rotameter type or a soap bubble with a nylon brush, and rinse in the hot detergent solu-
type is satisfactory. Airflow should be measured at each tion. Rinse thoroughly with tap water, followed by
tube under the conditions of the test and should be adjust- deionized water, and place upright in a test tube rack.
ed to 2.5 ± 0.1 mL/sec. Alltech provides a digital flow Fill with deionized water and soak at least 1 hr. At the
meter, Model 7080, suitable for air flow measurement. end of the soaking period, rinse the tubes once again
7. Sample reaction tubes—soda glass 25 mm × 200 mm. with fresh deionized water, drain on clean filter paper
Each tube requires a two-hole stopper and aeration and dry. Store in a dust-free location.

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 SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 12b-92 • Oil Stability Index

Tube cleaning, alternative procedure—Add excess 2. Allow the oil temperature to equilibrate for 15 min
potassium hydroxide (KOH) to an isopropyl before reading the thermometer. When noting the tem-
alcohol/water solution (90:10, v/v) and mix until satu- perature, do not move the thermometer up or down.
ration has occurred. Half fill the tubes with this solu- 3. Adjust the block temperature controller to compensate
tion, and heat in a boiling water bath for several hours for any differences. Block temperature must be within
with occasional shaking. Finally, rinse with deionized ± 0.1°C of the target temperature (e.g., for 110°C in the
water. If traces of oil remain, the saponification must range of 109.9–110.1°C; for 130°C in the range of
be repeated. When the tubes have been cleaned in this 129.9–130.1°C).
manner, they should be immersed in a 1% solution of 4. After adjustments have been made, wait until the con-
detergent at >80°C for 1 hr. Finally, rinse all glassware troller is showing the new temperature, and then allow
with deionized water and dry. the oil to equilibrate for 15 min. Record the tempera-
3. Conductivity tube and probe cleaning—To clean the ture and repeat calibration steps 2–4 until the target
conductivity tubes and probes, first rinse the reaction temperature is reached.
products with denatured ethanol. The hot detergent 5. Check the temperature periodically to ensure accuracy.
solution should then be used as a soak or, with the aid
of a brush, to scrub conductivity probes that have the PROCEDURE
rigidity to withstand such treatment. Rinse repeatedly 1. Fill the conductivity tubes with 50 mL deionized water
with deionized water, and soak the tubes and probes in and attach the probes. Verify that the water conductivi-
deionized water for at least 1 hr. When ready to reuse, ty in the tube is 25 µS-cm−1 or less and that the con-
fill the tubes with deionized water and connect the ductivity is constant.
probe to the conductivity meter. After 30 min, if the 2. Unless already completely liquid, the sample should be
water conductivity is under 25 µS-cm −1 and is not melted at a temperature not more than 10°C above its
changing, then the conductivity tube is ready for reuse. melting point. The sample size is 5.0 ± 0.2 g for OSI
If the water conductivity in the tube is too high (i.e., and Rancimat instruments without an insert, or 2.5 ±
>25 µS-cm−1), the probe must be recleaned. 0.2 g for Rancimat instruments fitted with disposable
inserts. Carefully place the sample directly into the
SAMPLING bottom of the reaction tube. It has been found that bet-
1. Because this procedure is a measure of oil stability, ter precision is obtained if the sample is not allowed to
anything that might detrimentally affect the stability of coat the side of the tube. Avoid, as noted in the sam-
the sample must be avoided. Samples should be kept pling procedure, contaminating the sample during
cool and in the dark. Where packaged fats are transfer.
involved, the sample should consist of an unopened 3. In the collaborative studies, all determinations were
package, if possible. If this is not possible, samples performed at 110 and 130°C. The temperature should
must be removed from large containers or processing be selected so as not to permit the OSI time to be less
equipment with clean sampling devices of stainless than 4 hr or more than 15 hours. Times less than 4 hr
steel, aluminum, nickel or glass. Samples of solid fat result in a wider variation of end point determination.
should be taken at least 5 cm from the walls of large Sample temperature should be checked periodically to
containers and 2.5 cm from the walls of small contain- ensure the temperature controllers have not drifted.
ers. If liquid oil is poured from a container, the pouring 4. Connect the tubing from the air manifold to the con-
spout or lip should first be cleaned thoroughly using a ductivity measurement tube, and adjust the aeration
clean cloth moistened with acetone. After removal tubes to within 5 mm of the bottom of both the reac-
from packages or processing equipment, samples tion and the conductivity tubes. Measure the airflow,
should be transported and stored only in clean glass or adjusting to 2.5 ± 0.2 mL/sec (see Note, 3). An exhaust
plastic containers. Samples should be protected from hood is beneficial for the removal of obnoxious
contact with heat, light and air as much as possible. volatiles resulting from oxidation reactions.
Samples should be stored with little or no headspace; if 5. A computer or multichannel strip chart recorder should
headspace is unavoidable, purge the headspace with be used to monitor the conductivity of each probe in
nitrogen. the instrument. A plot of water conductivity vs. time
obtained from the recorder is then analyzed, and the
TEMPERATURE CALIBRATION OSI inflection point is determined either by a micro-
Note—Because variations in temperature are potentially the processor-computed slope/change algorithm or a maxi-
most serious source of error, temperature calibration of the mum of the second derivative, or by the tangential
instrument is critical. method (see Fig. 2), etc.
1. The oil sample should be added to the reaction tube to
the level equivalent to 5 g for OSI and Rancimat PRECISION
instruments without an insert, and 2.5 g for Rancimat 1. A collaborative study, in which the stability of rape-
instruments fitted with disposable inserts. The airflow seed oil and palm oil was determined at 100°C by 11
is then adjusted as directed (Apparatus, 6), and a ther- laboratories in Norway and the UK, was reported by
mometer (Apparatus, 5) is suspended in the oil such Woestenburg and Zaalberg (References, 4) to have
that it does not touch the walls of the reaction tube. given an interlaboratory reproducibility coefficient of

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SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 12b-92 • Oil Stability Index

variation (RSD R) of 9.1% and an instrumental repeata- 6. Table 2 contains the statistical analysis of the interlab-
bility coefficient of variation of 3.3%. oratory study carried out by the AOCS A O M
2. The OSI method was subjected to a collaborative study Alternatives Committee in 1991, in which 14 laborato-
among 15 laboratories, using all currently available ries participated, each obtaining four test results for
commercial instruments noted in this method. The each sample analyzed at 130°C (statistical results eval-
overall average coefficient of variation was 10.2% for uated in accordance with ISO 5725-1986).
samples having from 7 to 80 hr of stability at 110°C.
Summaries of the statistical analysis of the results of NOTES
the collaborative study carried out by the AOCS Caution
Active Oxygen Method (AOM) Alternatives Trichloroethane is an irritant to eyes and skin. The TLV is
Committee are shown in Table 1 for 110°C and in 350 ppm in air.
Table 2 for 130°C. Generally, a larger variation was Acetone is highly flammable and forms explosive perox-
noted for lower temperatures and longer induction ides with oxidizing agents. Use effective fume-removal
periods. device. Do not mix with chloroform. The explosive limits in
3. Repeatability limit—The absolute difference between air are 2.6–12.8%. The TLV is 750 ppm. It is narcotic in high
two independent single test results, obtained with the concentrations and moderately toxic by ingestion and inhala-
same method on identical test material in the same lab- tion.
oratory by the same operator using the same equipment
within short intervals of time, should not be greater NUMBERED NOTES
than the repeatability limit (r) as calculated from the 1. The OSI analysis may be used to analyze many other
formulas in Tables 1 and 2. types of oils and fats; however, some of these were
4. Reproducibility limit—The absolute diff e r e n c e not included in the collaborative study. Extremely
between two single test results, obtained with the same stable fats may be analyzed by accelerating the rate of
method on identical test material in different laborato- oxidation by raising the temperature at which the oil
ries with different operators using different equipment, is oxidized (e.g., to 140°C). The analysis of these
should not be greater than the reproducibility limit (R) high-stability oils and fats can be performed if the
as calculated from the formulas in Tables 1 and 2. conductivity cell water loss is kept to a minimum by
5. Table 1 contains the statistical analysis of the interlab- cooling, or is kept at constant volume by the periodic
oratory study completed by the AOCS AOM addition of fresh water to make up for any loss. The
Alternatives Committee in 1991, in which 15 laborato- stability of free fatty acids has been found to be too
ries participated, each obtaining four test results for low to permit analysis at 110°C, but the analysis
each sample analyzed at 110°C [statistical results eval- could be performed at a lower temperature. Fish oils
uated in accordance with International Organization and linseed oils have been successfully analyzed at
for Standardization (ISO) 5725-1986]. 80°C, but in general have a long, sloping oxidation

Table 1
Statistical results of interlaboratory study in which samples were analyzed at 110°C.
Samplea
A B C D E F I J L
Number of
labs after
removal of
outliers 15 14 15 15 13 14 13 12 13
n 58 55 56 57 48 52 49 45 51
Outliers 0 3 1 1 8 4 0 4 0
Mean, hoursb 10.1 7.7 17.9 23.7 13.9 46.5 68.1 69.7 18.6
Repeatability, Sr 0.6 0.4 0.8 0.9 0.3 1.2 4.2 3.5 1.8
RSDr 5.96 4.80 4.34 4.00 2.11 2.51 6.22 5.03 9.62
r (2.8 × Sr) 1.68 1.12 2.24 2.52 0.8 3.36 11.76 9.80 5.04
Reproducibility, SR 1.3 0.9 2.0 1.9 1.14 4.1 13.5 9.0 4.4
RSDR 12.33 11.23 11.09 8.13 8.18 8.92 19.88 12.92 23.93
R (2.8 × SR) 3.64 2.52 5.60 5.32 3.19 11.48 37.91 25.23 12.32
aKey to samples: A, sunflower/soybean oil blend; B, nonhydrogenated soybean oil; C, liquid/hydrogenated soybean oil blend; D, hydro-
genated soybean oil/cottonseed oil blend; E, liquid/hydrogenated corn oil blend; F, hydrogenated corn oil/cottonseed oil blend; I, jojoba
oil; J, high-stability oil; L, crude corn oil.
bInduction period in hours.

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 SAMPLING AND ANALYSIS OF COMMERCIAL FATS AND OILS
Cd 12b-92 • Oil Stability Index

Table 2
Statistical results of interlaboratory study in which samples were analyzed at 130°C.
Samplea
A C D E F I J
Number of
labs after
removal of
outliers 14 14 14 14 14 11 14
n 51 51 53 54 50 41 49
Outliers 1 0 2 2 3 4 4
Mean, hoursb 2.6 4.4 5.8 3.4 10.9 11.5 16.1
Repeatability, Sr 0.13 0.18 0.18 0.13 0.46 0.39 0.60
RSDr 4.98 4.14 3.13 3.85 4.24 3.44 3.70
r (2.8 × Sr) 0.36 0.50 0.50 0.36 1.29 1.09 1.01
Reproducibility, SR 0.32 0.49 0.54 0.39 1.09 1.74 1.70
RSDR 12.26 11.04 9.27 11.37 10.01 15.17 10.51
R (2.8 × SR) 0.90 1.37 1.51 1.09 3.05 4.87 4.76
a Key to samples: A, sunflower/soybean oil blend; C, liquid/hydrogenated soybean oil blend; D, hydrogenated soybean oil/cottonseed oil
blend; E, liquid/hydrogenated corn oil blend; F, hydrogenated corn oil/cottonseed oil blend; I, jojoba oil; J, high-stability oil.
b Induction period in hours.

curve rather than a sharply defined rapid rise. ple onto the walls of the reaction tube, it may be helpful
Margarines and mayonnaises may be tested after sep- to decrease the size of the bubbler orifice (References, 6).
aration and drying of the oil with anhydrous magne-
sium sulfate (MgSO 4) prior to analysis. REFERENCES
2. The presence or absence of moisture in the air may 1. deMan, J.M., F. Tie and L. deMan, J. Am. Oil Chem.
have an effect on the oxidative stability of certain Soc. 64:993 (1987).
antioxidants (References, 5). As noted in Reference 5, 2. Jebe, T., M.T. Matlock and R.T. Sleeter, J. Am. Oil
when analyzing fats containing moisture-susceptible Chem. 70:1055 (1993).
antioxidants, it is advisable to place a drying tower 3. Matlock, M.G., T. Jebe and R.T. Sleeter, Unpublished.
containing a suitable drying agent in the air train. 4. Wo e s t e n b u rg, W.J., and J. Zaalberg, Fette Seifen
3. Precision decreases if the oil coats the side of the reaction Anstrichm. 88:53 (1986).
tube. If the required airflow causes splattering of the sam- 5. Mehlenbacher, V.C., The Analysis of Fats and Oils,

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