Food Anal.

Methods (2012) 5:14531458

DOI 10.1007/s12161-012-9399-1

A New Method for Determining Free Fatty Acid Content

in Edible Oils by Using Electrical Conductivity
Xiuzhu Yu & Chen Yang & Shuangkui Du & Jin-ming Gao

Received: 7 November 2011 / Accepted: 8 March 2012 / Published online: 24 March 2012
# Springer Science+Business Media, LLC 2012

Abstract In the current study, a new method for determining free fatty acid (FFA) content in edible oils has been
developed based on the electrical conductivity (EC) value
change of a 0.04 M potassium hydroxide (KOH) solution
during KOHFFA mixed reaction. At ambient temperature,
4.0000.001 g of edible oils (oils with a high melting point
were melted first) was dispersed in 40.00.1 ml of KOH
solution. The mixtures were stirred for 5 s and were held for
5 min. The EC value changes of the KOH solution layers were
determined, and the FFA was predicted basing on the calibration. The analytical conditions studied include the concentration of KOH solution, holding time, the types of edible oils,
setting temperature, and the ratio of oil to KOH solution (m/v).
The calibration was validated by using the American Oil
Chemists Society (AOCS) titrimetric FFA method. The analytical conditions were found to have no significant effects
on the results. When the ratio of oil to KOH solution was
1:10 (m/v), the calibration was y012.04x0.0769 (where x is
the EC value change and y is the FFA content), R00.9992,
SD00.0745, and a calibration with an SE of <0.0745 % FFA
over a range of 0 % to 5 % was obtained. The method was
validated using standard addition and the AOCS method. The
results show that the analytical performance of the EC
X. Yu (*) : C. Yang : S. Du
College of Food Science and Engineering,
Northwest A&F University,
28 Xinong Road Yangling,
712100 Shaanxi, China
e-mail: xiuzhuyu1004@hotmail.com
J.-m. Gao (*)
College of Science, Northwest A&F University,
22 Xinong Road Yangling,
712100 Shaanxi, China
e-mail: jinminggaocn@yahoo.com.cn

procedure is as good as or better than that of the AOCS

titrimetric procedure.
Keywords Electrical conductivity . Edible oils . Free fatty
acid . Analysis

Introduction
Fats and oils from a wide variety of sources are important to
the food industry and other industrial sectors, and their free
fatty acid (FFA) content is an important quality parameter.
The determination of FFA is important for edible oil and
biodiesel processes and quality control analyses (Hamm and
Hamilton 2000), which is used to monitor deodorizer efficiency (OBrien 1998) and serve as indicators for frying oil
replacement (Senorans and Ibanez 2002). The determination
of FFA by using the standard method of the American Oil
Chemists Society (AOCS) involves titrating oil dissolved
in alcohol with a strong base to a phenolphthalein end point
(AOCS 1989). Although these methods are simple, they are
tedious and problematic, especially when dealing with dark
oils. In addition, these methods require large amounts of
solvents that could pose potential environmental threats.
Therefore, various alternative approaches have been developed to measure FFA, including flow injection methods
(Nouros et al. 1997; Takamura et al. 1999; Mariotti and
Mascin 2001), as well as pH metric (Kwon and Rhee 1986)
and colorimetric techniques (Turyan et al. 1996). Chromatographic techniques, such as gas, high-performance liquid and
supercritical fluid chromatography, have also been used for
determining FFA (Dermaux et al. 1999; Kotani et al. 2002;
Senorans and Ibanez 2002; Rosenfeld 2002). These methods
provide more information at the expense of introducing more
complexity into the analysis. Fourier transform infrared

1454

(FTIR) spectroscopy has also been investigated as a means of

measuring FFA in edible oils. FTIR spectroscopy is a simple,
rapid, and non-destructive technique, which plays a critical
role in the rapid determination of various parameters of fats
and oils (Alberta et al. 2009; Yu et al. 2007). A large number
of studies on the determination of FFA contents in vegetable
oil have been published (Alberta et al. 2009; Al-Alawi et al.
2004, 2005). Sherazi et al. (2007, 2009) developed methods
using single-bound attenuated total reflectance FTIR spectroscopy to assess the different quality parameters of edible oils.
FTIR methods are limited by their expensive instruments and
are unsuitable for in-plant quality control or centralized commercial laboratories. The sensitivity of these methods is insufficient for catering to the wide range of edible oil quality. On
the other hand, other methods are lengthy and make use of
toxic organic solvents, which not only increase the cost of the
analysis, but also are the sources of potential health risks and
environmental pollution.
Electrical conductivity (EC) is a measure to test the
capability of a material to conduct an electric current. EC
is a very useful property because its values are affected by
the chemical composition of substances and the stress state
of crystalline structures. Therefore, EC information can be
used to estimate the amount of total dissolved salts (Giancoli
2007). The concentration of anions and cations, where H+,
K+, and OH are the most important, determines the EC
value changes in the potassium hydroxide solution during
the FFA and potassium hydroxide (KOH) reactions. Zhu et
al. (2008) developed a method for detecting hogwash oil
adulterated in edible oil by measuring the EC value of the
oils. The developed calibration and validation showed that
EC was a practicable and simple method for identifying
edible oil adulteration.
The standard method commonly used for FFA analysis is
based on the titration of a sample of oil with a KOH solution to
a phenolphthalein end point. In this study, we established an
effective method based on the EC value changes in a potassium hydroxide solution during reaction, and the developed
calibration was validated.

Materials and Methods

Food Anal. Methods (2012) 5:14531458

status. All chemicals and reagents used were of analytical

grade and were purchased from Xian Chemical company,
Ltd. (Xian, China).
Instrumentation
An EC instrument, DDSJ-308A, from Shanghai Precision &
Scientific Instrument Co., Ltd., was used.
Determination Principle
The underlying principle of FFA determination is based on
the EC value changes of the mixtures (oilKOH solutions)
during reaction. When the temperature is constant, the EC
value depends on the concentration of the KOH solution.
After neutralizing the FFA by using excessive KOH, its EC
decreased with the reduction of the concentration of KOH
solution. Then, the concentration of the KOH solution and
the changes in EC value were used to develop the calibration and to determine FFA in edible oils. The KOH solution
should be in a low concentration to avoid the saponification
of the oils.
Optimization Studies
Optimization studies on the effects of KOH solution concentration, holding time, the different types of edible oils,
and the ratio of oil to KOH solution (m/v), and setting
temperature were performed. The adopted experimental
conditions were a compromise between the analytical conditions and the EC value changes.
Effect of KOH Concentration
At ambient temperature, different concentrations of KOH solutions were prepared at approximately 0.01, 0.02, 0.03, 0.04,
0.05, and 0.06 M, and the EC of the solutions were determined
respectively. Then, 4.000 g of edible oils were dispersed in
40.0 ml of KOH solution. The mixtures were stirred for approximately 5 s and held for 5 min. The EC values of the KOH
solution layers were determined, and the EC value changes
were calculated. The oil layer was analyzed using a titration
method to ensure the complete reaction of the FFA.

Reagents
Effect of Holding Time
Various edible oils were obtained from local retail outlets.
Among these oils, refined rapeseed oil was used as the base
oil for preparing the calibration standards, whereas the other
oils include blind oil samples that were obtained from a
local market. After the FFA-free oil was acquired, it was
passed through a column of microwave-activated silica gel
to remove any polar oxygenated molecules, and the AOCS
Ca 5a-40 procedure was performed to confirm FFA-free

At ambient temperature, the EC value of the KOH solutions

(about 0.04 M) was determined. Then, 4.000 g of different
edible oils were dispersed in 40.0 ml of KOH solution. The
mixtures were stirred for approximately 5 s and held at
different time points ranging from 5 to 35 min with 5-min
intervals. The EC values of the KOH solution layers were
determined, and the EC value changes were calculated.

Food Anal. Methods (2012) 5:14531458

Effect of Different Types of Edible Oils

At ambient temperature, the EC value of the KOH solutions
(about 0.04 M) was determined. Then, 4.000 g of different
edible oils were dispersed in 40.0 ml of KOH solution. The
mixtures were stirred for approximately 5 s and held for
5 min. The EC values of the KOH solution layers were
determined, and the EC value changes were calculated.
The regression equations between FFAs and EC value
changes were developed for each oil type and compared
with each other.
Effect of Setting Temperature
The effect of setting temperature on the EC valued was
determined by adding 4.000 g of soybean oil to 40 ml of
0.04 M KOH solution at various temperatures: 15, 20, 25,
30, 35, 40, 50, 60, 70, 80 C, stirring for about 5 s and
holding at the specific temperature for 5 min. Palm oil
samples were also studied at 50, 60, 70, and 80 C by
adding melted sample, stirring, and holding. After setting,
the EC values of the KOH solution were determined.

1455

obtained by using the EC procedure. A calibration equation

for the prediction of FFA content was derived by using the
linear regression of the gravimetrically determined FFA
percentages to the EC value changes.
Validation
A number of validation samples were spiked with hexanoic
acid, whereas others, which contain different levels of FFA,
were collected from the local market. The value changes of
the standards were obtained using the EC procedure. The
validation samples were processed by using the calibration
equation, and the reported FFA results were compared with
the added hexanoic acid (expressed as a percentage of oleic
acid) and the values obtained by using the AOCS titrimetric
FFA (AOCS Ca 5a-40) method immediately prior to the EC
analysis.
Statistical Analysis
Analysis of variance and Duncans multiple range tests were
performed using the SAS program version 9.1 for Windows.
Statistical significance was set at p<0.05.

Effect of Ratio of Oil to KOH Solution

At ambient temperature, the EC value of KOH solutions
(about 0.04 M) was determined. Then, 4.000 g of different
edible oils (containing different levels FFA) were dispersed
in 8.0, 12.0, 16.0, 20.0, and 40.0 ml of KOH solution. The
mixtures were stirred for approximately 5 s and held for
5 min. The EC values of the KOH solution layers were
determined, and the EC value changes were calculated.
The regression equations between FFAs and EC value
changes were developed for each ratio of oil to KOH solution (m/v) and compared with each other.
Analytical Protocol
At ambient temperature, prior to EC analysis, about 0.04 M
KOH solution was prepared, and its EC value was determined. Then, 4.0000.001 g of standard and validation
samples (oils with high melting point were melted first)
was dispersed in 40.00.1 ml of KOH solution. The mixtures were stirred for approximately 5 s and were held for
5 min. The EC values of the KOH solution layers were
determined, and the EC value changes were calculated.
Calibration
Calibration standards were prepared through the gravimetric
addition of hexanoic acid to the FFA-free rapeseed oil to
span an FFA range of 0 % to 5 % (expressed as a percentage
of oleic acid). The EC value changes of the standards were

Results
Analytical Condition
Effect of KOH Concentration
The KOH concentration was chosen from 0.01 to 0.06 M to
investigate the effect of KOH concentration on EC value
changes. It showed that the change of EC with molarity was
very small. When the KOH concentration varied between
0.02 and 0.06 M, the EC value changes ranged from 0.060
0.002 S/m to 0.0650.001 S/m. Statistical analysis showed
that the KOH concentration had no significant (P>0.05)
effect on the EC value changes. The FFA content in the oil
layer was also determined based on the AOCS titrimetric
FFA method; the oil layer did not have detectable FFA.
Therefore, KOH concentrations from 0.02 to 0.06 M can
be used for different requirements under various conditions,
and 0.04 M was chosen for the following experiments.
Effect of Holding Time
The KOH was mixed thoroughly in soybean oil, rapeseed
oil, and peanut oil samples with different levels of FFA. The
solution was held, and the EC value changes of the KOH
solution layers were then determined from 5 to 35 min with
5-min intervals. When the holding time varied from 5 to
35 min, the ranges of the EC value changes for three

1456

Food Anal. Methods (2012) 5:14531458

treatments were 0.0430.002 S/m to 0.0450.002 S/m,

0.033 0.002 S/m to 0.0350 0.002 S/m, and 0.019
0.001 S/m to 0.0200.001 S/m, respectively. Plots of EC
versus holding time showed that slopes of the three treatments were 4105, 6105, and 6105; they were all
close to zero. Statistical analysis showed that the holding
time had no significant (P>0.05) effects on the EC change
values. Thus, the holding time of 5 min was sufficient for
the current experiment and was chosen for the analysis.

Effect of Ratio of Oil to KOH Solution

The plots of FFA to the EC value changes with the various
ratios of oil to KOH solution were linear and fit the following equations:
y1:2 2:63x 0:053 R 0:9997; SD 0:008

y1:3 3:58x 0:063 R 0:9995; SD 0:009

y1:4 5:93x 0:004 R 0:9993; SD 0:023

y1:5 9:60x 0:111 R 0:9990; SD 0:012

y1:10 11:85x 0:093 R 0:9995; SD 0:009

Effect of Different Types of Edible Oils

Different types of edible oils have different fatty acid compositions and the presence of different ions, which may
affect the experiment. Different levels of FFA in rapeseed,
olive, coconut, and palm oils were determined, and the EC
value changes were calculated prior to EC analysis (Fig. 1).
Figure 1 demonstrates good linearities between different
levels of FFA in the oils and EC value changes, with high R
values (R>0.99), and the slopes and intercepts of the equations made from olive, palm, coconut, and rapeseed oils are
very close. No effect seems to be associated with the oil
types.
Effect of Setting Temperature

FFA (%)

Plots of the EC values for soybean oil and palm oil at

various temperatures were shown to have little effect on
the EC values. The range of EC change values for soybean
oil and palm oil at various temperatures from 15 to 80 C and
50 to 80 C were 0.0650.002 S/m to 0.0670.002 S/m, and
0.1000.001 S/m to 0.9970.001 S/m, respectively. Statistical analysis showed that the temperatures had no significant
(P>0.05) effect on the EC value changes. Temperature effects
could be ignored in the EC procedure, so we chose ambient
temperature for the FFA analysis.

These equations show that as the ratio of oil to KOH

solution decreases, the slopes of the equations increase, and
the intercepts that give the best fit of the data vary slightly.
The range of determination was wider, but the precision
decreased with the elevation of the ratio of oil to KOH
solution. For an edible oil sample, low EC value changes
yield larger errors. All of the R values exceeded 0.99, and all
of the ratio of oil to KOH solution were variable. Different
ratios of oil to KOH solution can be used in different experiments basing on the needs of the range and the precision of
determination. A ratio of oil to KOH solution of 1:10 (m/v)
was used in the current study.
Calibration

0.8

Reliable results for free fatty acid content were verified in

the range of 0 % to 5 % using a ratio of oil to KOH solution
of 1:10. The results could be predicted by the following
equation:

0.7

y 12:04x  0:0769; R 0:9992; SD 0:0745;

0.6

where x is the EC value change and y is the FFA content

(percent).
In the range of 0 % to 5 % FFA, a high R value and the
overall SD of 0.0745 % show that the performed calibrations give highly consistent results.

0.5
Olive oil
y=12.44x-0.0158, R=0.9998,SD=0.0473
Palm oil
y=11.82x-0.0323, R=0.9935,SD=0.0303
Coconut oil
y=12.50x-0.0166, R=0.9995,SD=0.0109
Rapeseed oil
y=12.71x-0.0136, R=0.9991,SD=0.0136

0.4
0.3
0.2
0.1

0.0
0.00 0.01 0.02 0.03 0.04 0.05 0.06 0.07 0.08 0.09

EC value changes (S/m)

Fig. 1 FFA versus EC plots

Validation
Validation samples of different types of edible oils were
prepared by spiking fresh oils (0 % FFA as determined by
the AOCS titrimetric method) with hexanoic acid in the
amount required to cover 0 % to 5 %. The EC FFA was
calculated using Eq. 6. The following regression equation

Food Anal. Methods (2012) 5:14531458

1457

Table 1 Results of the triplicate analysis using the AOCS method and
the EC method
Number

AOCS method

EC method

Mean

SD

Mean

SD

1
2
3
4
5
6
7
8
9
10

0.457
0.130
0.136
1.419
1.864
1.964
2.054
3.214
4.029
4.892

0.036
0.053
0.029
0.124
0.044
0.065
0.086
0.068
0.154
0.126

0.467
0.156
0.297
1.431
1.810
1.523
1.946
3.226
4.102
4.825

0.057
0.058
0.014
0.019
0.024
0.037
0.048
0.053
0.097
0.087

Mean SDr
SDDa

0.078
0.042

0.049
0.027

Results expressed as the percentage of oleic acid (w/w)

SDr SD for reproducibility, SDDa SD of the differences for accuracy

was obtained by using the AOCS and EC methods for the oils
that were subjected to the standard addition of FFA mixture
relative to the gravimetrical data:
y 0:933x 0:0591; R 0:9995; SD 0:0471;

where x is the added FFA and y is the EC FFA (percent).

The regression equation illustrates the excellent concurrence between the titrimetric and EC methods, as well as
their comparable capability to track the amounts of FFA
gravimetrically added to the oils. The results also indicate
a close to correspondence 1:1 between the methods and
show that the intercepts are close to zero. Thus, the methods
concur to within 0.0471 %. According to Eq. 7, no effect
seems to be associated with oil type. The regression SD
shows that the relative sensitivity of the EC method was
approximately 0.1 % FFA content. The EC method is workable for the FFA analysis in edible oils.
Blind Oil Samples Validation
To further validate the performance of the EC method,
ten different edible oil samples were collected from the
local market. The oils were immediately analyzed using
the AOCS titrimetric FFA method in triplicate, with
titrations being completed as quickly as possible before
EC analysis. The obtained results compared with the
analytical data provided with these oil samples are presented in Table 1.
Generally, an excellent concurrence between the EC means
and FFA means was obtained using the AOCS method, and the
EC method was shown to be capable of producing accurate

and reproducible FFA data independent of the oil type. Moreover, the EC method appears to be a valid alternative to the
AOCS titrimetric procedure. In terms of overall reproducibility, the EC method had a mean SDr of 0.049 %, which was
slightly better than that of the AOCS procedure (0.078 %). For
each method, the value of SDDa, which is a measure of the
variability around the MDa, is of the same order of magnitude
with the SDr value, where the EC method again performed
slightly better.

Discussion and Conclusion

The current study aimed primarily to simplify FFA
analyses in edible oils by the application of the EC
procedure. Results showed that the EC method is comparable to the AOCS Ca 5a-40 method in terms of
accuracy. Moreover, the procedures of the EC method
are easier to perform than that of the AOCS method.
Moreover, the KOH solution need not be prepared in
accurate concentration. However, earlier procedures using large quantities of organic solvents lead to environmental pollution. Detection range and accuracy depend
on EC value changes, and the EC value relates to the
ratios of sample oil to KOH solution. The method
becomes scalable with a simple adjustment in the oil/
solution ratio, and the analysis of edible oils containing
about 0.1 % FFA is possible. The method has a strong
sensitivity, which makes the use of FFA as an indicator
of lipid oxidation possible. Furthermore, the sensitivity
of this method accurately monitors the refining process
conducted in edible oils. The method can easily be
automated when combined with automation devices.
The study showed that experimental parameters had no
significant effect on the determination of FFA. Different
ratios of oil to KOH solution can be used under different
conditions depending on the range and precision. When the
ratio of oil to KOH solution was 1:10 (m/v), the calibration
was y012.04x0.0769, R00.9982, SD00.0745. The results
from two methods were highly consistent, and the relative
deviation of the data on the blind samples experiment was
less than 10 %. This method avoids the subjectivity from
judging the end point and does not require any organic
solvent. Moreover, the EC instrument is cheap and affordable for general QC laboratories. With these hallmarks of a
sound general method, this new methodology could serve as
an alternative to the AOCS instrumental method for the
analysis of FFA in edible oils.
Acknowledgments The authors would like to thank the China Postdoctoral Science Foundation (20110491694) and the Fundamental
Research Funds for the Central Universities (QN 2009074) for the
financial support.

1458

References
Al-Alawi A, van de Voort FR, Sedman J (2004) New FTIR
method for the determination of FFA in oils. J Am Oil Chem
Soc 81:441446
Al-Alawi A, van de Voort FR, Sedman J (2005) A new FTIR method
for the analysis of low levels of FFA in refined edible oils.
Spectrosc Lett 38:389403
Alberta A, van de Voort FR, Simpson BK (2009) FTIR determination
of free fatty acids in fish oils intended for biodiesel production.
Process Biochem 44:401405
AOCS Method Ca 5a-40, Official methods and recommended practices
of AOCS, 4th edn, AOCS, Champaign, IL, 1989
Dermaux A, Sandra P, Ferraz V (1999) Analysis of free fatty acids and
fatty acid phenacyl esters in vegetable oils and margarine by
capillary electro-chromatography. Electrophoresis 20:7479
Giancoli D (2007) Physics for scientists and engineers (4th edn.)
mastering physics series. Upper Saddle River, NJ: Prentice Hall
Hamm W, Hamilton RJ (2000) Edible oil processing. Sheffield Academic Press, Sheffield, p 234
Kotani A, Kusu F, Takamura K (2002) New electrochemical detection
method in high-performance liquid chromatography for determining free fatty acids. Anal Chim Acta 465:199206
Kwon DY, Rhee JS (1986) A simple and rapid colorimetric method for
determination of free fatty acids for lipase assay. J Am Oil Chem
Soc 63:186189
Mariotti E, Mascini M (2001) Determination of extra virgin olive oil
acidity by FIA-titration. Food Chem 73:235238

Food Anal. Methods (2012) 5:14531458

Nouros PG, Georgiou CA, Polissiou MG (1997) Automated flow
injection spectrophotometric non-Aqueous titrimetric determination of the free fatty acid content of olive oil. Anal Chim Acta
351:291297
OBrien RD (1998) Fats and oils: formulating and processing for
applications. Technomic Publishing Company, Lancaster, p 96
Rosenfeld JM (2002) Application of analytical derivatizations to the
quantitative and qualitative determination of fatty acids. Anal
Chim Acta 465:93100
Senorans FJ, Ibanez E (2002) Analysis of fatty acids in foods by
supercritical fluid chromatography. Anal Chim Acta 465:131141
Sherazi STH, Mahesar SA, Bhanger MI, van de Voort FR, Sedman J
(2007) Rapid determination of free fatty acids in poultry feed lipid
extracts by SB-ATR FTIR spectroscopy. J Agric Food Chem
55:49284932
Sherazi STH, Talpur MY, Mahesar SA, Kandhro A, Arain S (2009)
Main fatty acid classes in vegetable oils by SB-ATR-Fourier
transform infrared (FTIR) spectroscopy. Talanta 80:600606
Takamura K, Fuse T, Arai K, Kusu F (1999) A review of a new voltammetric method for determining acids. J Electroanal Chem 468:5363
Turyan YI, Berezin OY, Kuselman I, Shenhar A (1996) pH-metric
determination of free fatty acids in vegetable oils without titration.
J Am Oil Chem Soc 73:295301
Yu X, van de Voort FR, Sedman J (2007) Deternination of proxide
value of edible oils by FTIR spectroscopy with the use of the
spectral reconstitution technique. Talanta 74:241246
Zhu R, Wang D, Yang XJ, He DP (2008) Application of electrical
conductivity measurement in qualified edible oil adulteration
identification. J Cereals & Oils 11:4243