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Comparison of OXITEST and RANCIMAT methods to evaluate the oxidative

stability in frying oils

Article  in  European Food Research and Technology · October 2017

DOI: 10.1007/s00217-017-2995-y

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Eur Food Res Technol
DOI 10.1007/s00217-017-2995-y
ORIGINAL PAPER
Comparison of OXITEST and RANCIMAT methods to evaluate
the oxidative stability in frying oils
Federica Tinello1 · Anna Lante1 · Michele Bernardi2 · Francesca Cappiello2 ·
Fernanda Galgano3 · Marisa Carmela Caruso3 · Fabio Favati2   
Received: 20 July 2017 / Revised: 22 September 2017 / Accepted: 13 October 2017
© Springer-Verlag GmbH Germany 2017
Abstract  The oxidation of fatty acids in the presence of
atmospheric oxygen is recognized as one of the main fac-
tors affecting the shelf life of oils and fats causing rancidity
by the formation of off-flavors due to aliphatic aldehydes or
other volatile compounds. Therefore, the oxidative stability
is one of the most important parameters to be evaluated in
the formulation of commercial frying oils. In this regard, the
induction period of 15 frying oils has been measured using
two different methods of accelerated oxidation, OXITEST
and RANCIMAT, and correlated to their fatty acid content
previously detected by gas chromatographic technique.
Frying oils containing high contents of saturated fatty acids
mainly from the palm oil achieved an improved oxidative
stability by increasing their induction period. Moreover,
the linear regression analysis showed a good correlation
between the induction period values of the two instruments.
Hence, the innovative OXITEST method may be an easy,
fast, and eco-friendly alternative to the official RANCIMAT
method for evaluating the oxidative stability in oil- and fat-
containing products.
All authors contributed equally to this work.
* Fabio Favati
fabio.favati@univr.it
1
Department of Agronomy, Food, Natural Resources,
Animals, and Environment‑DAFNAE, Agripolis, University
of Padova, Viale Università 16, 35020 Legnaro, PD, Italy
2 stability of frying oils. Some measurements of the oxi-
Department of Biotechnology, University of Verona, Strada
Le Grazie 15, 37134 Verona, Italy
3 properties and volatile and non-volatile decomposition
School of Agricultural, Forestry, Food and Environmental
Sciences, University of Basilicata, Viale dell’Ateneo Lucano
10, 85100 Potenza, Italy
Keywords  Oxidative stability · Induction period ·
RANCIMAT · OXITEST · Frying oils · Fatty acids
Introduction
Frying, which is one of the most commonly used cook-
ing procedures in household, restaurants, fast-food chains,
and food industry, is a system involving oil, food, fryer and
processing [1]. The deep- and shallow-frying techniques
include, respectively, the dipping or contact of food with
hot oil at the temperature range of 150–190 °C to increase its
organoleptic quality and shelf life [2]. The hydrolysis, oxi-
dation and polymerization reactions occurring in the heated
oil during frying lead to the formation of non-volatile and
volatile compounds which affect the sensory and nutritional
properties not only of the oil but also of the fried food. These
chemical reactions depend on several factors such as frying
temperature and time, oxygen concentration, the presence
of antioxidants, and the type of oil and fryer [3]. The frying
stability of oils is associated mainly to their tocopherol and
fatty acid contents and profiles [4–6]. Mixtures of differ-
ent vegetable oils containing saturated and unsaturated fatty
acids are recommended in the formulation of frying oils. In
fact, the inclusion of oils rich in saturated fatty acids such as
palm oil in mixtures of unsaturated fatty acid-containing oils
can reduce the formation of oxidized products because of the
faster oxidation of the unsaturated rather than the saturated
fatty acids [7, 8].
There are several methods to evaluate the quality and
dation degree in frying oils are focused on oil physical
products [9]. Other methods which measure the resist-
ance to oxidation in forced conditions take into account
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 Eur Food Res Technol

the oxidative stability as one of the most important factors
to evaluate the shelf life, palatability, nutritional quality
and toxicity of oils and fats by determining the induction
period (IP), a parameter representing the time needed to
reach the starting point of lipid oxidation. Therefore, a
longer IP can be associated with a higher oxidation stabil-
ity. The active oxygen method (AOM) and the analysis of
carbonyl value (CV) have been the mostly used tests to
detect the oxidative stability of oils by measuring primary
or secondary oxidation products, such as hydroperoxides,
aldehydes and ketones, respectively [10]. However, these
chemical techniques, which are non-reproducible, time
consuming, expensive and involve the use of toxic rea-
gents, have been then replaced with accelerated oxidation
methods [11]. In this regard, the official oil stability index
(OSI) method of American Oil Chemists’ Society (AOCS)
requires the use of different analytical instruments that
can be utilized as an alternative, RANCIMAT (Metrohm,
Herisau, Switzerland) being one of these [12]. The RAN-
CIMAT method allows to evaluate not only the oxidative
stability of edible [13] and non-edible oils [14], fats [11],
nanoemulsions [15], encapsulated extract within nanoe-
mulsion and multiple emulsions [16] but also the antioxi-
dant performance of plant extracts [17, 18] by monitor-
ing over time the water conductivity, which increases as a
consequence of volatile acids formed after heating samples
under atmospheric pressure and constant values of temper-
ature and air flow. The OXITEST—Oxidation Test Reactor
(VELP, Usmate, MB, Italy)—is an innovative instrument
that can be utilized to investigate the oxidation stability
of solid [19–21] or liquid [22, 23] samples, without prior
preparation. The instrument monitors the oxygen uptake
of the reactive components present in food/feed samples to
evaluate the oxidative stability under accelerated oxidation
conditions, and the reactor temperature can be regulated
from room temperature to 110 °C, while the oxygen pres-
sure can reach up to 0.8 MPa. A few papers have investi-
gated its use for evaluating the oxidation stability of oils,
chia seeds, and bakery products [20, 22–25] and recently
the AOCS has approved a new analytical procedure (Cd
12c-16) based on the use of this instrument.
Aim of this work was to compare the OXITEST and
the RANCIMAT methods in evaluating the oxidative sta-
bility of commercial frying oils, taking also into account
the relationship between the IP values and the fatty acid
composition of the oils.
Materials and methods
Samples
A total of 15 frying oils from different commercial brands
were purchased from a local market and stored at room
temperature under dark conditions until the analyses. All
of the commercial oils were manufactured and packaged
in Italy with the exception of sample D, coming from Bel-
gium (Table 1).

Table 1  Composition and fat Code Composition Fat (g/100 mL)

content declared in the label of
the frying oil samples analysed
in the study
A Sunflower oil high in linoleic acid, sunflower oil high in oleic acid, cori- 92.0
ander essential oil (0.0002%)
B Sunflower, peanut oils 100a
C Sunflower, palm, peanut oils 92.0
D Sunflower oil, rapeseed oil, sunflower oil high in oleic acid, corn oil, 92.0
dimethylpolysiloxane (anti-foaming agent)
E Sunflower, fractionated vegetable oils NR
F Sunflower, fractionated palm oils 91.6
G Sunflower, fractionated palm oils 91.4
H Sunflower, fractionated palm oils 91.4
I Sunflower, fractionated palm, peanut oils 91.6
J Sunflower, fractionated palm, peanut oils 92.0
K Sunflower, soybean, fractionated palm oils 92.0
L Sunflower, bi-fractionated palm, peanut oils 92.0
M Sunflower, bi-fractionated palm, peanut oils 91.4
N Sunflower, bi-fractionated palm, peanut oils 91.6
O Sunflower (70%), bi-fractionated palm (20%), soybean (5%) oils 92.0

NR value not reported in the label

a
  g/100 g

13
Eur Food Res Technol
Oxidative stability measurement using the RANCIMAT
instrument
The oxidative stability of the frying oils was evaluated using
a RANCIMAT apparatus (Metrohm, model 743, Herisau,
Switzerland) and measuring over time the water conductiv-
ity [18]. A fixed amount of oil (3.0 ± 0.1 g) was added to
each reaction tube and subjected to accelerated oxidation
by raising the temperature to 110 °C under a 20 L h−1 air
flow. The volatile oxidation products formed under forced
conditions were transported by the air stream into the con-
ductometric cell, previously filled with 60 mL of distilled
water, and detected by continuously measuring the water
electrical conductivity (µS cm−1) over time (h). The oxi-
dative stability was expressed as the IP corresponding to
the time (h) at the intersection point between the horizontal
(conductivity, µS min−1) and vertical (time, h) tangents of
the fitted exponential oxidation curves. At this break point
the water conductivity increased over time because of the
fat oxidation. Two samples per frying oil were placed in the
equipment and analysed simultaneously in triplicate.
Oxidative stability measurement using the OXITEST
instrument
The oxidative stability of the frying oils was also assessed
using the OXITEST apparatus (­ Velp® Scientifica, Usmate,
Italy), featuring two thermostated and hermetically sealed
titanium chambers, each one able to contain up to three
round 35-mL titanium sample holders, that can be piled up
when working with large sample amounts. When the sample
size requires the use of less than three holders, the unused
ones are replaced with titanium spacers, so to keep constant
the void volume in each chamber [22]. An amount equal to
10 g of frying oil, thoroughly mixed immediately before
being sampled, was loaded in each chamber and due to the
limited sample amount only one holder was utilized, being
then necessary the use of two spacers. The working tempera-
ture was set at 110 °C, while the initial ­O2 pressure (grade
5.0; SAPIO, Monza, Italy) was set at 0.6 MPa. The instru-
ment was controlled by a specific software (OXISoft™,
­Velp® Scientifica, Usmate, Italy) that allowed to measure
over time the absolute pressure change in the two chambers,
monitoring the oxygen uptake of the active components of
the samples. For each chamber at the end of the test, the
program automatically calculated the IP from the resultant
oxidation curve, by means of a graphical method (two tan-
gent methods). Each analysis was repeated three times for a
total of six data acquired for each oil.
Assessment of the oil fatty acid content
The saturated and unsaturated fatty acid content in the fry-
ing oils was assessed by gas chromatographic analysis of
the respective methyl esters (FAMEs) [26]. The analyses
were carried out using an Agilent 7890A GC system (Agi-
lent Technologies, Cernusco sul Naviglio, Milano, Italy)
equipped with an Agilent 7683 autosampler and a Flame
Ionization Detector (FID). FAMEs were prepared by weight-
ing 40 mg of each frying oil in a small glass test tube and
adding 2 mL of hexane and 200 µL of 2 M methanolic KOH.
The mixture was then vortexed at room temperature for
2 min and then let to stand after the addition of about 0.5 g
of anhydrous ­Na2SO4 to eliminate any water formed during
the hydrolysis process. After the phase separation occurred,
an aliquot of the upper hexane layer (1 μL) was injected
in split mode (1:50) into a S­ LB®-IL111 capillary column
(100 m × 0.25 mm × 0.20 μm) (Sigma-Aldrich, Milano,
Italy). Helium (grade 6.0, SOL, Monza, Italy) was used as
carrier gas (1 mL min−1), while nitrogen (grade 5.0, SOL,
Monza, Italy) was used as make-up gas. The injector and
FID temperatures were set at 250 °C and separation of the
various FAMEs was achieved using an initial oven tempera-
ture of 100 °C, which after being kept constant for 13 min
was then raised to 240 °C at 4 °C min−1. The various fatty
acids were identified according to their retention time and
by spiking the samples, using a commercial mixture of pure
standards (Supelco 37 Component FAME Mix CRM47885,
Sigma-Aldrich, Milano, Italy). The fatty acid content was
expressed as percentage based on the sum of the detected
peak areas.
Statistical analysis
The data concerning the IP values and fatty acid content
were analysed by a one-way analysis of variance (ANOVA),
after verifying the normal distribution and homogeneity of
variance, using the PROC GLM of Statistical Analysis Sys-
tem (SAS) [27]. The model included the type of frying oil as
fixed effect. Least-square means were chosen for presenting
all data and compared using the Tukey’s multiple range test.
Differences among means with P ≤ 0.05 were accepted as
representing statistically significant differences. The box-
plots were computed using the SAS software package [27].
Linear regression analysis was carried out using the
PROC REG of SAS [27]. The OXITEST IPs were correlated
with those obtained using the RANCIMAT instrument, and
the assessed IPs of both instruments were correlated with
the fatty acid content.
13
 Eur Food Res Technol

Results and discussion Furthermore, other studies have shown that when using the
OXITEST the working temperature may affect the oxygen
Correlation between RANCIMAT and OXITEST solubility in lipids, with a reduction of the assessed IP val-
ues of about 25% for each 10 °C increment [22]. Anyhow it
The frying oils consisted of edible mixtures derived from should also be pointed out that when using the RANCIMAT
two or three different vegetable oils achieving a total fat apparatus the oil sample was exposed to a continuous flow of
content around 92 g/100 mL (Table 1). As regards the com- air at atmospheric pressure, having an oxygen concentration
position declared in the label of the frying oils analysed in of about 21%. Conversely, when working with the OXITEST
this study, sunflower oil was present in all samples. Moreo- instrument the oil sample was exposed to pure ­O2 under
ver, both A and D frying oils contained sunflower oil high pressure (0.6 MPa).
in oleic acid, while only the former contained also sunflower The OXITEST and RANCIMAT instruments achieved
oil high in linoleic acid. The enrichment of oleic [4] and the same results in spite of their different measurement
linoleic acids [5] in sunflower oil leads to health benefits, approach. In this regard, the effect of the type of frying
improved shelf life and fast formation of polar compounds oil on the IP value was statistically significant (P ≤ 0.001)
during frying. The D sample from Belgium contained also regardless of the analytical method applied. The boxplots in
rapeseed and corn oil, and dimethylpolysiloxane as anti- Fig. 1 divided the frying oil samples into three main groups
foaming agent to further protect lipids from thermal deg- based on their IP values obtained using both the OXITEST
radation [28], while the A sample contained also coriander and the RANCIMAT methods. In the first group the K and N
essential oil (0.0002%) to improve the flavor. Palm oil was samples, with IP mean values of about 5.42 and 10.47 h for
present in most of the investigated frying oils and the F–K OXITEST and RANCIMAT methods, respectively, showed
and L–O samples contained the fractionated and bi-frac- the highest oxidative stability (Table 2). In the second group,
tionated forms of palm oil to improve the oxidative stability the C–J and L–M samples, with IP mean values of about
and shelf life of the products [29]. Peanut oil was found in 3.73 and 7.00 h for OXITEST and RANCIMAT methods,
the composition of 7 out of the 15 studied oils (B, C, I, J
and L–N samples) but reasonably at different percentages,
being reported in the labels at the second place for the B
sample and at the third place for the others. Only K and O
samples contained soybean oil as second and third ingredi-
ent, respectively, while the E oil composition resulted to be
not well defined, being only reported in the label that the
second of the two ingredients was represented by “fraction-
ated vegetable oils”.
The oxidative stability of the frying oil samples was eval-
uated by measuring their IPs with two different methods.
In particular, with the RANCIMAT instrument, where the
accelerating oxidation factors are temperature and air flow,
the IP values were assessed as the time needed for achieving
an increase in the measured water conductivity due to the
production of oxidation-related moieties. Conversely, with
the OXITEST method, where the accelerating factors are
temperature and high levels of pure ­O2, the IP values rep-
resented the time needed to achieve a significant pressure
decrease of the oxidizing gas in the chamber, thus a sudden
change in the lipid oxidation rate.
To compare the two instruments at similar analytical
conditions, the working temperature was set at 110 °C for
both, taking into account that temperature can play a cru-
cial role in fat oxidation, therefore, affecting the determina-
tion of the IP values. In particular, some authors studying
the relationship between RANCIMAT and active oxygen
method at different temperatures have reported an oxidative
stability decrease of about 27% for sunflower, palm, soybean Fig. 1  Boxplots of the IP values of the frying oils obtained by OXIT-
and rapeseed oil when moving from 110 to 130 °C [11]. EST (a) and RANCIMAT (b) methods

13
Eur Food Res Technol

Table 2  IP values of the frying oils obtained by OXITEST and

RANCIMAT methods
Sample IP OXITEST (h) IP RANCIMAT (h)
Mean ± SD CV% Mean ± SD CV%

A 2.69i ± 0.08 2.97 4.78h ± 0.26 5.36

B 2.79i ± 0.05 1.79 4.80h ± 0.15 3.08
C 3.61fg ± 0.15 4.16 6.72fg ± 0.08 1.21
D 3.85cde ± 0.15 3.90 7.74c ± 0.20 2.52
E 4.07bc ± 0.08 1.97 7.43cd ± 0.38 5.11
F 3.93bcd ± 0.02 0.51 7.35cd ± 0.16 2.19
G 3.79def ± 0.08 2.11 6.84ef ± 0.13 1.90
H 3.64efg ± 0.15 4.12 6.66fg ± 0.19 2.81 Fig. 2  Correlation between the IP values of the frying oils obtained
I 3.51g ± 0.12 3.42 7.03def ± 0.13 1.85 by OXITEST and RANCIMAT methods
J 3.89bcd ± 0.05 1.28 7.15de ± 0.11 1.57
K 5.38a ± 0.11 2.04 10.67a ± 0.21 1.98
L 3.72defg ± 0.11 2.96 6.74ef ± 0.09 1.32 E, the CV% values resulted to be higher than 5%, specifically
M 3.27h ± 0.10 3.06 6.31g ± 0.22 3.53 5.11 and 5.36% (Table 2).
N 5.46a ± 0.09 1.65 10.27a ± 0.27 2.65
O 4.11b ± 0.16 3.89 9.03b ± 0.12 1.29 Correlation between oxidative stability and fatty acid
content
SD standard deviation, CV% coefficient of variation
a–h
   Values are the mean (±  standard deviation) of three replicates.
Means within each column and with different superscripts are statisti- The oxidative stability is affected by several factors includ-
cally different (P ≤ 0.05) ing the content of saturated and unsaturated fatty acids [4],
tocopherols [5] and other antioxidants compounds [15] in
the oil samples and the storage conditions [30]. The fatty
respectively, showed an intermediate oxidative stability. In acid profile of oils is generally recognized as the most deci-
the third group, the oils A and B, with IP mean values of sive parameter influencing the oxidation stability of oils [8].
about 2.74 and 4.79 h for OXITEST and RANCIMAT meth- Moreover, some authors highlighted that the frying stability
ods, respectively, showed the lowest oxidative stability. of high oleic sunflower oil is influenced by the fatty acid
Only for sample O, the IPs assessed with the two methods content rather than the tocopherol isomeric composition
resulted in a different ranking of the oil as regards its oxida- [5]. Therefore, the oxidative stability of frying oils has been
tion stability. In particular, when tested with the OXITEST discussed on the basis of their fatty acid content which was
method its IP was not significantly different from those of determined by GC analysis. As reported in Table 3 all frying
samples E, F and I, oils with an intermediate relative oxi- oil samples were rich in unsaturated and poor in saturated
dative stability. Conversely, when using the RANCIMAT fatty acids with a total mean of 83.7 and 16.3%, respec-
instrument its IP resulted more similar to those of oils K and tively. Moreover, the fatty acid content differed significantly
N, characterized by the highest stability (Table 2). (P ≤ 0.001) based on the formulation of frying oils. In detail,
Statistical analysis of the oils IP data assessed with the the boxplot in Fig. 3 divided the frying oil samples into three
OXITEST and the RANCIMAT methods revealed a signifi- groups based on their saturated fatty acid content: (1) K
cant positive correlation (r = 0.97; r2 = 0.93; P ≤ 0.001) and N samples high in saturated fatty acids whose content
between the IP values of these different methods (Fig. 2). was on average of 27.3%; (2) C, E–J, and O samples with
In detail, the IPs obtained when using the OXITEST were an averaged content of saturated fatty acids around 17.3%;
approximately two times lower than those obtained when (3) A, B, D, L, and M samples low in saturated fatty acids
using the RANCIMAT instrument. As a consequence, the whose content was on average 10.3% (Table 3). The boxplot
OXITEST method was comparable to the RANCIMAT one in Fig. 4 divided the frying oil samples into three groups
for evaluating the relative oxidative stability of the frying based on their unsaturated fatty acid content: (1) A, B, D, L,
oils but it accelerated more the fat oxidation, thus detecting and M samples high in unsaturated fatty acids whose content
the IP value in a shorter time. In addition, both methods was on average of 89.7%; (2) C, E–J, and O samples with an
showed good CV% values, mainly below 5%. In particular, averaged content of unsaturated fatty acids around 82.7%;
for OXITEST the assessed CV% values ranged from 0.51 to (3) K and N samples low in unsaturated fatty acids whose
4.16, while for RANCIMAT the values ranged from 1.21 to content was on average of 72.7% (Table 3). The boxplot in
3.53. However, with the latter instrument for two oils, A and Fig. 5 divided the frying oil samples into three groups based

13
 Eur Food Res Technol

Table 3  Fatty acid content of the frying oils

Sample Saturated fatty acids (%) Unsaturated Saturated/unsatu-
fatty acids (%) rated fatty acids

A 10.13h ± 0.03 89.87bc ± 0.03 0.11g ± 0.00

B 11.38g ± 0.01 88.63  ± 0.01 0.13fg ± 0.00
c

C 15.65e ± 0.18 84.36e ± 0.18 0.19e ± 0.01

D 7.45i ± 0.66 a
92.56  ± 0.66 0.08a ± 0.01
E 19.88b ± 0.05 80.12h ± 0.05 0.25b ± 0.00
F 18.32c ± 0.10 81.69  ± 0.10 0.23bc ± 0.01
g

G 17.63c ± 0.14 82.38g ± 0.14 0.22cd ± 0.01

H 16.03de ± 0.36 83.97ef ± 0.36 0.20de ± 0.01
I 17.09cd ± 0.02 82.92fg ± 0.02 0.21cde ± 0.00
J 16.18de ± 0.59 83.82ef ± 0.59 0.20de ± 0.01
K 27.44a ± 0.04 72.57i ± 0.04 0.38a ± 0.00
L 9.65h ± 0.08 b
90.35  ± 0.08 0.11g ± 0.00 Fig. 4  Boxplot of the unsaturated fatty acid content of the frying oils
M 12.98f ± 0.31 87.02d ± 0.31 0.15f ± 0.00
N 27.23a ± 0.03 i
72.77  ± 0.03 0.37a ± 0.00
O 17.55c ± 1.09 82.45  ± 1.09 0.22cd ± 0.02
g

a–h
   Values are the mean (±  standard deviation) of three replicates.
Means within each column and with different lowercase letters are
statistically different (P ≤ 0.05)

Fig. 5  Boxplot of the ratio between saturated and unsaturated fatty

acid content of the frying oils

and unsaturated fatty acid content (Figs. 6, 7, respectively).

Fig. 3  Boxplot of the saturated fatty acid content of the frying oils
on their ratio between saturated and unsaturated fatty acid
content (Table 3): (1) K and N samples with an averaged
ratio of 0.38; (2) C, E–J, and O samples with an averaged
ratio of 0.22; (3) A, B, D, L, and M samples with an aver-
aged ratio of 0.12.
A comparison between the results of the IP and fatty acid
content suggested that the frying oils with greater IP values
had a higher saturated fatty acid content. Several authors
have reported that fat oxidation decreases at increasing
levels of saturated fatty acids and at decreasing levels of
unsaturated fatty acids [4, 8, 21]. In this regard, the IP val-
ues of the frying oil samples obtained using OXITEST and
RANCIMAT methods have been correlated to their saturated
The correlation has also been investigated between the IP
values and the ratio between saturated/unsaturated fatty
acids (Fig. 8) to assess the relationship between the fatty
acid composition and the measured oxidative stability.
The positive correlation of the IP values obtained using
OXITEST (r = 0.83; r2 = 0.69; Fig. 6a) and RANCIMAT
(r = 0.78; r2 = 0.61; Fig. 6b) methods with the saturated
fatty acid content was statistically significant (P ≤ 0.001).
The IP values increased at increasing saturated fatty acid
content. In detail, each 10% increment of the saturated fatty
acid content in the frying oils extended approximately 1
and 2 h the IPs obtained with OXITEST and RANCIMAT
methods, respectively. The negative correlation of the IP
values obtained using OXITEST (r = − 0.837; r2 = 0.69;
Fig. 7a) and RANCIMAT (r = − 0.78; r2 = 0.61; Fig. 7b)
methods with the unsaturated fatty acid content was statisti-
cally significant (P ≤ 0.001). The IPs decreased at increasing

13
Eur Food Res Technol
Fig. 6  Correlation of the IP values of the OXITEST (a) and RANCI-
MAT (b) methods with the saturated fatty acid content of the frying
oils
Fig. 7  Correlation of the IP values of the OXITEST (a) and RANCI-
MAT (b) methods with the unsaturated fatty acid content of the fry-
ing oils
saturated fatty acid content. In detail, each 10% increment of
the unsaturated fatty acid content in the frying oils decreased
of approximately 1 and 2 h the IP obtained with OXITEST
and RANCIMAT methods, respectively. The positive cor-
relation of the IP values obtained using OXITEST (r = 0.86;
r2 = 0.73; Fig. 8a) and RANCIMAT (r = 0.80; r2 = 0.64;
Fig. 8b) methods with the ratio saturated/unsaturated fatty
acid content was statistically significant (P ≤ 0.001). The
IPs resulted to increase at increasing saturated/unsaturated
ratios, with a 10% increment of the latter causing roughly an
increase of approximately 0.8 and 1.5 h of the IPs assessed
Fig. 8  Correlation of the IP values of the OXITEST (a) and RANCI-
MAT (b) methods with the ratio between saturated and unsaturated
fatty acid content of the frying oils
with the OXITEST and the RANCIMAT instruments,
respectively. Hence, an increased oxidative stability of the
frying oils was related to an increase in the saturated fatty
acid content based on the formulation of the ingredients
declared in the label (Table 1). In details, the frying oils K
and N, which showed to be the most stable according to their
assessed IPs (5.4 and 5.5 h with OXITEST; 10.7 and 10.3 h
with RANCIMAT) (Table 2), were the richest in saturated
fatty acids, (27.4 and 27.2%, respectively) (Table 3) whose
levels may be associated to the use of fractionated palm oil
in the industrial formulations (Table 1). In fact palmitic acid
was found to be present in oils K and N at the highest levels,
22.4 and 22.5%, respectively. Moreover, the fractionation
process applied on the palm oil utilized in these samples
(Table 1) led to a greater oxidative stability in comparison
to the oil C, containing unfractionated palm oil and char-
acterized by a lower palmitic acid level (10.9%) and lower
IP values, as assessed with the OXITEST and RANCIMAT
methods (3.61 and 6.72 h, respectively) (Table 2). On the
contrary, samples A and B, that resulted to be more prone
to oxidation (Fig. 1) according to their lowest IP values
assessed using the OXITEST (2.69 and 2.79 h) or the RAN-
CIMAT (4.78 and 4.80 h) method, did not contain any palm
oil and showed to be poor in saturated fatty acids (10.1 and
11.4%, respectively) and rich in unsaturated fatty acids (89.9
and 88.6%), due to their composition based on sunflower and
peanut oils (Tables 1, 2, 3). Several authors have reported
that palm oil, rich in palmitic acid, has shown to be less sen-
sitive to oxidation than other vegetable oils rich in unsatu-
rated fatty acids and olive oil [7, 29, 31]. Furthermore, some
authors evaluating the oxidative stability of several vegetable
oils with the RANCIMAT method at different temperatures,
13

confirmed the greatest resistance of palm oil to fat oxida-
tion, being characterized by higher IP values (19.95, 10.89,
and 5.70 h) than rapeseed (7.38, 4.10, and 1.85 h), soybean
(5.24, 2.61, and 1.51 h) and sunflower oils (3.70, 1.89, and
0.88 h) at 110, 120, and 130 °C, respectively [11].
Conclusions
The detection of IP value represents an important parameter
for evaluating the oxidative stability of frying oils. Acceler-
ated tests such as OXITEST and RANCIMAT are able to
speed up the fat oxidation and monitor the oxidative stability
in a relatively short period of time without using expen-
sive and environmental hazardous reagents. The regression
analysis carried out on the IP values showed a linear correla-
tion between RANCIMAT and OXITEST instruments which
achieved the same comparative results following different
procedures. Hence, the OXITEST method may represent
a valid alternative to the RANCIMAT method and could
be a useful tool for the edible oil industry in formulating
oil mixtures to be utilized for food frying. Furthermore, the
method does not necessitate any specific sample preparation
and requires less time for carrying out oxidative stability
studies, therefore, allowing higher throughputs.
Acknowledgements  The authors would like to thank VELP Scienti-
fica (Usmate, MB, Italy) for lending the OXITEST instrument and Dr.
Paola Ornaghi for technical assistance.
Compliance with ethical standards  18. Mihaylova DS, Lante A, Tinello F, Albert IK (2014) Study on
Conflict of interest  The authors declare that they have no conflict
of interest.
Compliance with ethics requirements  This article does not contain
any studies with human or animal subjects.
References
1. Stier RF (2004) Frying as a science—an introduction. Eur J Lipid
Sci Technol 106:715–721
2. Oreopoulou V, Krokida M, Marinos-Kouris D (2006) In: Mujum-
dar AS (ed) Handbook of industrial drying, 3rd edn. CRC Press,
Boca Raton
3. Choe E, Min DB (2007) Chemistry of deep-fat frying oils. J Food
Sci 72:77–86
4. Marmesat S, Morales A, Velasco J, Dobarganes MC (2012) Influ-
ence of fatty acid composition on chemical changes in blends
of sunflower oils during thermoxidation and frying. Food Chem
135:2333–2339
5. Aladedunye F, Przybylski R (2013) Frying stability of high oleic
sunflower oils as affected by composition of tocopherol isomers
and linoleic acid content. Food Chem 141:2373–2378
13
Eur Food Res Technol
6. Gliszczyńska-Swigło A, Sikorska E, Khmelinskii I, Sikorski M
(2007) Tocopherol content in edible plant oils. Pol J Food Nutr
Sci 57:157–161
7. De Marco E, Savarese M, Parisini C, Battimo I, Falco S, Sac-
chi R (2007) Frying performance of a sunflower/palm oil blend
in comparison with pure palm oil. Eur J Lipid Sci Technol
109:237–246
8. De Leonardis A, Macciola V (2012) Heat-oxidation stabil-
ity of palm oil blended with extra virgin olive oil. Food Chem
135:1769–1776
9. Melton SL, Jafar S, Sykes D, Trigiano MK (1994) Review of
stability measurements for frying oils and fried food flavor. J Am
Oil Chem Soc 71:1301–1308
10. AOCS (1989) Official methods and recommended practices of the
AOCS, 4th edn. AOCS, Champaign, Ill
11. Anwar F, Bhanger MI, Kazi TG (2003) Relationship between ran-
cimat and active oxygen method values at varying temperatures. J
Am Oil Chem Soc 80:151–155
12. AOCS (2012) Official methods and recommended practices of the
AOCS, 6th edn. AOCS, Champaign, Ill
13. Mateos R, Uceda M, Aguilera MP, Escuderos ME, Maza
GB (2006) Relationship of Rancimat method values at vary-
ing temperatures for virgin olive oils. Eur J Lipid Sci Technol
223:246–252
14. Peer MS, Kasimani R, Rajamohan S, Ramakrishnan P (2017)
Experimental evaluation on oxidation stability of biodiesel/die-
sel blends with alcohol addition by rancimat instrument and FTIR
spectroscopy. J Mech Sci Technol 31:455–463
15. Lante A, Friso D (2013) Oxidative stability and rheological prop-
erties of nanoemulsions with ultrasonic extracted green tea infu-
sion. Food Res Int 54:269–276
16. Mohammadi A, Jafari SM, Esfanjani AF, Akhavan S (2016)
Application of nano-encapsulated olive leaf extract in controlling
the oxidative stability of soybean oil. Food Chem 190:513–519
17. Lante A, Nardi T, Zocca F, Giacomini A, Corich V (2011) Evalu-
ation of red chicory extract as a natural antioxidant by pure lipid
oxidation and yeast oxidative stress response as model systems. J
Agric Food Chem 59:5318–5324
the antioxidant and antimicrobial activities of Allium ursinum L.
pressurised-liquid extract. Nat Prod Res 28:2000–2005
19. Riciputi Y, Caboni MF (2017) Assessing oil oxidative stability in
Tarallini by ­OXITEST®. Ital J Food Sci 29:63–73
20. Caruso MC, Galgano F, Colangelo MA, Condelli N, Scarpa T,
Tolve R, Favati F (2017) Evaluation of the oxidative stability of
bakery products by OXITEST method and sensory analysis. Eur
Food Res Technol 243:1183–1191
21. Verardo V, Riciputi Y, Sorrenti G, Ornaghi P, Marangoni B,
Caboni MF (2013) Effect of nitrogen fertilisation rates on the
content of fatty acids, sterols, tocopherols and phenolic com-
pounds, and on the oxidative stability of walnuts. Food Sci Tech-
nol 50:732–738
22. Comandini P, Verardo V, Maiocchi P, Caboni MF (2009) Acceler-
ated oxidation: comparative study of a new reactor with oxidation
stability instrument. Eur J Lipid Sci Technol 111:933–940
23. Caruso MC, Galgano F, Scarpa T, Ornaghi P, Favati F (2017)
Accelerated shelf life studies of extra virgin olive oils using the
Oxitest method. Inform 28:26–29
24. Verardo V, Riciputi Y, Trivisonno MC, Marconi E, Caboni MF
(2010) Effect of the addition of air-classified barley flours on
the lipid stability of bakery products. Eur Food Res Technol
231:309–319
25. Amato M, Caruso MC, Guzzo F, Galgano F, Commisso M, Boch-
icchio R, Labella R, Favati F (2015) Nutritional quality of seeds
and leaf metabolites of Chia (Salvia hispanica L.) from Southern
Italy. Eur Food Res Technol 241:615–625
Eur Food Res Technol

26. Ichihara K, Shibahara A, Yamamoto K, Nakayama T (1996) An
improved method for rapid analysis of the fatty acids of glyc-
erolipids. Lipids 31:535–539
27. SAS (2013). SAS/STAT(R) 9.2 User’s guide, 2nd edn. SAS Insti-
tute Inc., Cary. http://support.sas.com/documentation/cdl/en/
statug/63033/HTML/default/viewer.htm#glm_toc.htm. Accessed
29 June 2017
28. Gerde JA, Hammond EG, White PJ (2011) Influence of polydi-
methylsiloxane on the oxygen concentration of oils at various
temperatures. J Am Oil Chem Soc 88:925–929
29. Mba OI, Dumont MJ, Ngadi M (2015) Palm oil: processing, char-
acterization and utilization in the food industry—a review. Food
Biosci 10:26–41
30. Mancebo-Campos V, Salvador MD, Fregapane G (2014) Anti-
oxidant capacity of individual and combined virgin olive oil
minor compounds evaluated at mild temperature (25 and 40 °C)
as compared to accelerated and antiradical assays. Food Chem
150:374–381
31. Romano R, Giordano A, Vitiello S, Le Grottaglie L, Musso SS
(2012) Comparison of the frying performance of olive oil and
palm superolein. J Food Sci 77:519–531

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