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Synergetic Use of Principal Component Analysis

Applied to Normed Physicochemical


Measurements and GC × GC-MS to Reveal
the Stabilization Effect of Selected Essential Oils
on Heated Rapeseed Oil
Lilia Sghaier, Christophe B. Y. Cordella, Douglas N. Rutledge, Fanny Lefèvre, Mickaël Watiez, Sylvie Breton, Patrick Sassiat,
Didier Thiebaut, and Jérôme Vial

Abstract: Lipid oxidation leads to the formation of volatile compounds and very often to off-flavors. In the case of the
heating of rapeseed oil, unpleasant odors, characterized as a fishy odor, are emitted. In this study, 2 different essential oils

Food Chemistry
(coriander and nutmeg essential oils) were added to refined rapeseed oil as odor masking agents. The aim of this work was
to determine a potential antioxidant effect of these essential oils on the thermal stability of rapeseed oil subject to heating
cycles between room temperature and 180 °C. For this purpose, normed determinations of different parameters (peroxide
value, anisidine value, and the content of total polar compounds, free fatty acids and tocopherols) were carried out to
examine the differences between pure and degraded oil. No significant difference was observed between pure rapeseed
oil and rapeseed oil with essential oils for each parameter separately. However, a stabilizing effect of the essential oils, with
a higher effect for the nutmeg essential oil was highlighted by principal component analysis applied on physicochemical
dataset. Moreover, the analysis of the volatile compounds performed by GC × GC showed a substantial loss of the volatile
compounds of the essential oils from the first heating cycle.

Keywords: anisidine value, headspace volatiles, oxidative stability, peroxide value, rapeseed oil

Practical Applications: A solution to enhance the oxidative stability of rapeseed oil could be used to avoid the oxidation
of linolenic acid, which leads to the formation of off-flavors characterized by a fishy odor.

Introduction off-flavors characterized by a fishy odor (Hammer and Schieberle


Rapeseed oil is now the third vegetable oil, after soybean and 2013), which displeases consumers. It is therefore necessary to find
palm oils (Harwood and Gunstone 2007). This oil is consumed a solution to enhance the oxidative stability of rapeseed oil.
worldwide and its production represents more than 50 million One possibility to decrease the oxidation consists in the addi-
tons per year (Carré and Pouzet 2014). This oil provides essential tion of antioxidants into oils (Yanishlieva and Marinova 2001;
fatty acids, such as linoleic or linolenic acids. However, these Márquez-Ruiz and others 2014). The effect of substances on
compounds are subject to lipid oxidation, which leads to a decrease the stability of oils was characterized by different complemen-
in the nutritional and organoleptic properties of oils (Choe and tary methods allowing a monitoring of the degradation state of
Min 2006; Farhoosh and others 2009). Among the secondary oils during oxidation. The peroxide value (PV) is commonly used
oxidation products, some compounds are volatile and they can to measure the accumulation of hydroperoxides (Farhoosh and
affect flavor even at low concentrations due to their low odor others 2009). Various methods have been developed to perform
threshold (Kochhar 1996). More specifically, the oxidation of the this measure, based on the redox properties of hydroperoxides
linolenic acid present in rapeseed oil leads to the formation of (Barriuso and others 2013). Several secondary oxidation prod-
ucts can also be monitored. The anisidine value (AV) is re-
lated to the aldehydes generated during lipid secondary oxidation
JFDS-2016-1945 Submitted 11/23/2016, Accepted 3/18/2017. Authors Sghaier, (Barriuso and others 2013). The total polar compounds (TPC)
Lefèvre, Watiez, and Breton are with Lesieur, R&D Center ESPCI ParisTech - content is another marker of oils degradation. Polar compounds
CNRS, Coudekerque-Branche, France. Authors Sassiat, Thiebaut, Sghaier and Vial are produced as a result of the effect on lipids of atmospheric
are with Dept. of Analytical, Bioanalytical Sciences and Miniaturization (LSABM), oxygen, water present in the environment and a high tempera-
Inst. of Chemistry, Biology and Innovation (CBI) – ESPCI ParisTech, CNRS
ture (Farhoosh and others 2009). These compounds can be free
UMR 8231, PSL∗ Research Univ., 10 rue Vauquelin, 75231 Paris Cedex 05,
France. Authors Sghaier and Rutledge are with AgroParisTech, UMR1145 GENIAL fatty acids (FFA), diglycerides, oxidized triglyceride monomers,
Analytical Chemistry Laboratory, 16 rue Claude Bernard, 75005 Paris, France. triglyceride dimers, and triglyceride polymers (Dobarganes and
Author Cordella is with INRA, UMR1145 GENIAL Analytical Chemistry Lab., others 1988). Measurement of TPC, is directly related to the
16 rue Claude Bernard, 75005 Paris, France. Direct inquiries to author Sghaier degradation products present in oil, and this method is one of the
(E-mail: lilia.sghaier@gmail.com).
most accurate assessments of the thermo-oxidative degradation of
frying oils (Velasco and others 2004; Serjouie and others 2010;

C 2017 Institute of Food Technologists


 R

doi: 10.1111/1750-3841.13712 Vol. 82, Nr. 6, 2017 r Journal of Food Science 1333
Further reproduction without permission is prohibited
Effect of essential oils on rapeseed oil . . .

Joaquı́n Aladedunye and Przybylski 2013). Many countries have this issue, the use of chemometric tools can be considered. For
established legal limits for TPC in frying oils with a rejection point instance, principal component analysis (PCA) has already been
at 25% of TPC (Paul and others 1997; Matthäus 2006). Dimers successfully applied to data sets related to degraded oils (Cordella
and polymers are formed from oxidized triacylglycerols (TAGs) and others 2012; Zribi and others 2014).
and can provide indications of the oxidative stability of oils during The aim of this work was to propose an original approach to
heating (Aladedunye and Przybylski 2013, 2014). In accordance stabilize rapeseed oil with 2 different essential oils with a monitor-
with French law, the total amount of polymers of TAG must be less ing for 15 heating cycles. The oxidative stability at 180 °C of pure
than 14% (Tynek and others 2012). Another parameter often con- refined rapeseed oil and rapeseed oil containing nutmeg essential
sidered to evaluate the oxidative stability of frying oils for human oil (NEO) or coriander essential oil (CEO) was investigates and
consumption is the content of FFA. FFA result from the hydrol- compared. It should be noted that these essential oils were already
ysis of TAGs, but also from the decomposition of hydroperoxides used in commercial formulations by Lesieur. After several cycles
(Zribi and others 2014). Furthermore, tocopherols are important of heating, samples were examined with a set of complementary
constituents of oil and have nutritional properties (Robledo and analyses. Considering the complexity of the reaction occurring
others 2013). They also have an effect on the degradation rate of during frying or cooking, the choice has been made to work
oils as a result of their antioxidant activity (Normand and others without food in this study.
2001) and they can inhibit the formation of hydroperoxides, and
Food Chemistry

also reduce the formation of secondary oxidation products from Materials and Methods
the decomposition of hydroperoxydes (Lampi and others 1999; Unless otherwise specified, all reagents were purchased from
Kulås and others 2002). Consequently, the changes in the toco- Sigma Aldrich (Sigma-Aldrich Chemie, Saint-Quentin Fallavier,
pherols content are relevant concerning the degradation of the 3 France) and were of analytical or chromatographic grade.
studied formulations.
Among the secondary oxidation products, the analysis of volatile Oil samples
compounds is important because they are related to the deteriora- Refined rapeseed oil was supplied by Lesieur (Coudekerque-
tion of the flavor (Barriuso and others 2013). Gas chromatography Branche, France) and all experiments were performed on oil from
(GC) combined with headspace (HS) sampling is the technique the same batch.
of choice to analyze these molecules (Barriuso and others 2013). Three formulations were studied: non-enriched rapeseed oil
As a result of the complexity of the HS of edible oils, the use and rapeseed oil enriched with 10 ppm of NEO and 10 ppm of
of GC × GC is a real asset. Many studies to characterize the CEO.
volatile compounds from edible oils have been based on GC × GC NEO and CEO were purchased from René Laurent (Le Cannet,
(de Geus and others 2001; Hu and others 2014). Among HS tech- France).
niques, solid phase microextraction (SPME) is a selective approach
already successfully applied to the analysis of edible oils (Jeleń and Degradation procedure
others 2000; Doleschall and others 2003; Vichi and others 2003; Degraded oil samples were obtained by applying heating cycles
Guillen and Goicoechea 2008). in a fryer on 2.6 kg of oil. A heating cycle consisted in heating the
The oxidative stability of rapeseed oil has already been studied oil without food for 30 min from room temperature up to 180 °C,
(Kowalski and others 1997; Koski and others 2002; Joaquı́n Velasco then leaving it to cool down to 50 °C. Oil samples were taken
and others 2004). For example, Farhoosh and others determined at the end of each heating cycle (about 45 mL) with a headspace
the relation between different compositional parameters (PV, AV, in vials as small as possible. All samples were stored in the dark at
polyene index, total tocopherols content, total phenolics content, –20 °C until analysis. For practical reasons, only 1 sequence of
TPC) and the primary and secondary oxidation of a canola oil heating cycles was applied to each formulation.
heated at 180 °C (Farhoosh and Pazhouhanmehr 2009). Moreover,
several natural products are known for their antioxidant properties Oil analysis
(Yanishlieva and Marinova 2001). Urbančič and others (2014) Tocopherols. Tocopherols analyses were based on the ISO
showed a slowdown of the sunflower oil deterioration during 9936:2006 standard using a Shimadzu UFLC with a Prominence
deep-frying by the addition of rosemary extract. Ammari and RF-20A XS Shimadzu fluorescence detector (Shimadzu France,
others (2012a, 2012b) evaluated the antioxidant effect of a seed Marne la Vallée, France), set for excitation wavelength at 290nm
extract from Nigella sativa L. on corn and sunflower oils. Oils were and emission wavelength at 330 nm. The column was a normal-
heated at different temperatures between 120 and 190 °C and 3D- phase GenesisR
silica (250 mm x 4.6 mm; 4 μm; Grace Discovery
Front Face fluorescence analyses highlighted that the Nigella seed Sciences, Epernon, France). BHA (3-tert-butyl-4-hydroxyanisole)
extract improved the thermal stability and the shelf life of the oils. was used as internal standard. The concentrations of tocopherols
The antioxidant effect of several substances on rapeseed oil has also were expressed as milligrams for 100 g of oil. Only 1 determination
been evaluated. Taha and others (2014) examined the oxidative was performed for each sample, after analyzing 2 control samples.
stability of rapeseed oil treated with 3 different herbs: sage, thyme, Peroxide value. PV, expressed in milliequivalents of active
and rosemary. This study highlighted a higher oxidative stability oxygen per kilogram (mEq O2 /kg), is a measure of primary oxi-
of the treated oils at 120 °C, but a lower frying stability at 175 °C. dation products. PV was determined according to the method NF
Olmedo and others (2015) tested the antioxidant properties of EN ISO 3960 of June 2010. Potassium iodide was supplied by
oregano, laurel, and rosemary essential oils in sunflower oil. They PanReac AppliChem (PanReac AppliChem, Barcelona, Spain).
observed that these essential oils decreased the oxidative process in Only 1 determination was performed for each sample.
sunflower oil during accelerated oxidation. Anisidine value. AV is a measure of secondary oxidation prod-
The degradation of vegetable oils during oxidation is a com- ucts, more specifically unsaturated aldehydes. AV was determined
plex phenomenon and the interpretation of the results obtained according to NF EN ISO 6885. Absorbance was measured at
with different chemical analyses can be complicated. To overcome 350 nm using a Cary60 spectrophotometer (Agilent Technologies,

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Effect of essential oils on rapeseed oil . . .

Figure 1–Monitoring of polar


compounds during heating in
different formulations.

Food Chemistry
Figure 2–Monitoring of dimeric and
polymeric TAGs during heating in
different formulations.

Courtaboeuf, France). Experiments were performed in duplicate Volatile compounds. The characterization of the HS of
and as a final result the average was calculated. AV was expressed rapeseed oil was determined by HS-SPME-GC × GC-MS.
without units. A Triplus autosampler (Thermo-Electron Corporation,
Courtaboeuf, France) was used to carry out SPME. One milliliter
Free fatty acids. FFA content was determined by titration of
of oil sample was put in a 20 mL HS vial. The vial was sealed with
20 ± 0.01 g of rapeseed oil dissolved in 100mL of ethanol/ether
a Teflon/silicon septum and aluminum cap. A DVB/CAR/PDMS
(1:1, v/v), with 0.05 M sodium hydroxide solution, using thymol
50/30 μm 2-cm-long fiber was used to perform the extraction
blue (PanReac AppliChem, Barcelona, Spain) as indicator. FFA
of the volatile compounds from rapeseed oil in order to absorb
content was expressed as free oleic acid percentage. Only 1 measure
compounds with different polarities and volatilities. The SPME
was performed for each sample.
fiber was previously conditioned in the GC injector at 250 °C
Total polar compounds and dimeric and polymeric for 1 h following the manufacturer’s recommendations. The
triacylglycerols. TPC and dimeric and polymeric triacylglyc- vial containing the oil was preheated at 90 °C for 1 min. The
erols (DPTG) were determined by FT-NIR (Fourier Transform- SPME fiber was then exposed to the HS of the vial for 30 min
Near Infrared) spectroscopy using validated methods developed by at 90 °C. The SPME fiber was introduced into the inlet of the
Lesieur. Indeed, methods using NIR were recently developed to GC for 5 min at 250 °C to perform the volatile compounds
measure TPC and DPTG (Gertz and others 2013; Zribi and others desorption.
2014). An MPA multipurpose FT-NIR analyzer equipped with Analyses were performed on a Trace GC × GC gas chromato-
an OPUS LAB spectroscopy software interface (Bruker Optics, graph (Thermo-Electron Corporation, Courtaboeuf, France)
Marne la Vallée, France) was used for data analysis and acquisition. coupled with an ISQ quadrupole mass spectrometer (Thermo-
The TPC and DPTG contents were expressed as a percentage in Electron Corporation, Courtaboeuf, France). For separation, the
oil (weight %). Experiments were performed in duplicate and as a column set was a DB-5 (5% phenyl) primary column (30 m x
final result the average was calculated. 0.25 mm ID x 0.25 μm film thickness; Agilent Technologies,

Vol. 82, Nr. 6, 2017 r Journal of Food Science 1335


Effect of essential oils on rapeseed oil . . .
Food Chemistry

Figure 3–Monitoring of free fatty acids during heating in different formulations.

Figure 4–Changes in peroxide value during heating in different formulations.

Waldbronn, Germany) coupled with a DB-WAX column Statistical data treatment


(polyethylene glycol) (2 m x 0.1 mm ID x 0.1 μm film thickness; Estimation of methods variability. Before working on real
Agilent Technologies, Waldbronn, Germany). The oven temper- samples, all analyses, excepted analysis of volatiles, were replicated
ature was programmed from 35 °C, held for 3min, and raised to 6 times on a reference sample. This procedure aimed at estimat-
200 °C at 3 °C/min, then held for 4 min at this temperature. ing and controlling the standard deviations, which had to meet
The Trace GC × GC was equipped with a dual CO2 cryogenic the requirements defined by Lesieur for each analysis. The rela-
modulator. The modulation period was set at 7 s. Helium was the tive standard deviations obtained with these replicates were then
carrier gas at a constant flow of 1 mL/min. The injector tem- used to define the variability of the results for real samples with
perature was 250 °C. The temperatures of the ion source and these same analyses. This procedure allowed to remedy the limited
transfer line were 230 and 250 °C, respectively. Electron ioniza- number of replicates for each analysis.
tion mass spectra were recorded at 70 eV ionization energy. The It should be noted that the error bars shown in Figure 1 to
mass spectrometer operated in full scan mode, with an m/z range 6 represent the standard deviations obtained for 6 replicates of a
between 25 and 230 and an acquisition frequency of 44.5 Hz. reference sample.
Total ion current chromatograms were recorded using Xcalibur Analysis of variance. Analysis of variance (ANOVA) was
software (Thermo-Electron Corporation, Courtaboeuf, France). used to establish if the differences between the results obtained
The GC × GC representation was done using the Chrom-Card for pure oil, oil with NEO and oil with CEO were significant.
software (Thermo-Electron Corporation). One-way ANOVA was performed with a risk of first kind α =
Compounds were identified by comparison of their mass spectra 0.05 for each analysis using Microsoft Excel software (Microsoft).
with those of the mass spectral library NIST MS Search 2.0, and Principal components analysis. PCA is a chemometric tool
with retention time and mass spectra of standard compounds. for unsupervised data analysis (Cordella 2012). This technique

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Effect of essential oils on rapeseed oil . . .

Food Chemistry
Figure 5–Changes in anisidine value during heating in different formulations.

Figure 6–Changes in the α, γ , β and δ-tocopherol contents during heating in 3 different formulations.

reduces the dimensionality of numerical datasets in a multivariate and SAISIR R


Package (Cordella and Bertrand 2014) were used
problem. PCA is oriented towards modeling variance/covariance to perform statistical treatments.
structure of the data matrix into combinations of the original The matrix used to perform PCA was composed of 9 columns
variables which contains the significant variations. In this study, corresponding to the different analyses and 48 lines corresponding
PCA was applied to characterize the effect of different formu- to the 3 formulations degraded by fifteen heating cycles and the 3
lations on oil degradation. Matlab
R
2012 software (MathWorks) fresh oils. The data collected were centered and standardized.

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Effect of essential oils on rapeseed oil . . .
Food Chemistry

Figure 7–GC × GC chromatograms of fresh pure oil (A), pure oil after the first heating cycle (B), difference between fresh oil with NEO and fresh pure
oil (C), difference between oil with NEO and pure oil after the first heating cycle (D), difference between fresh oil with CEO and fresh pure oil (E), and
difference between oil with CEO and pure oil after the first heating cycle (F). The white circles show the characteristic compounds of EOs.

Results and Discussion Changes in the TPC and DPTG. As shown in Figure 1 and
The main focus of this work was to determine the effect of the 2, the total amounts of TPC and DPTG were of the order of 5%
addition of 2 different essential oils to rapeseed oil when heating and less than 3% in fresh oils, respectively.
at 180 °C. The formation of TPC and DPTG was linear and increased
with the number of heating cycles. The equations are reported in
Changes in the analytical indices Figure 1 and 2. At the end of 15 heating cycles, the formation of
The changes in TPC, DPTG, FFA, PV, AV, and tocopherols TPC did not exceed 25% for all samples. Interestingly, the TPC
content were evaluated for the 3 target formulations. All samples percentages were higher than 20% after 13 cycles for the oil with
were evaluated from 0 to 15 heating cycles. CEO, and after 14 cycles for pure oil, whereas the oil with CEO

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Effect of essential oils on rapeseed oil . . .

Figure 8–Scores (A) and loadings (B) for


PC1.

Food Chemistry

only reached this value after 15 cycles. Concerning the formation Changes in FFA. At the beginning of the heating experiment,
of DPTG, pure oil and oil with NEO reached the legal limit of all samples started with a low amount of FFA (<0.04%) (Figure 3),
14% after 14 heating cycles, whereas the amount of DPTG was which was common for refined oils (Normand and others 2001;
only 13.3% after 15 cycles in oil with CEO. Matthäus 2006).
However, the rates of TPC and DPTG formation were sim- After 15 heating cycles, the content of FFA did not exceed
ilar and no significant difference was observed between the 3 0.16%, which was still relatively low. The evolutions of FFA were
formulations. The evolution of TPC, and in particular the phe- not significantly different for the 3 formulations. These values were
nomenon of linear increase, was consistent with previous stud- lower than those observed in other studies (Normand and others
ies on rapeseed oil (Normand and others 2001; Matthäus 2006). 2001; Aladedunye and Przybylski 2014), but it could be due to
The amounts of DPTG observed were also logical with other the absence of introduction of food material into the oil, which
works about heated vegetable oils (Matthäus 2006; Zribi and would add water to the system and promote hydrolysis reactions
others 2014). (Zhang and others 2012).

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Effect of essential oils on rapeseed oil . . .

Figure 9–Scores (A) and loadings (B) for


PC2.
Food Chemistry

Changes in PV and AV. The initial PV and AV were less ondary oxidation products (Choe and Min 2006). For this reason,
than 0.2 mEq O2 /kg and 0.1, respectively, which indicated that the measurement of hydroperoxides (PV) is not always directly
fresh samples were not degraded before (Figure 4 and 5). correlated with sample oxidation, especially in the case of heated
The evolution of PV was characterized by a large increase be- samples (Casal and others 2010). The AV is an empiric measure-
tween the cycles 0 and 1, followed by a slight decrease until the ment of the content of aldehydes. Thus, this value is correlated
fourth cycle. After that, PV was more or less stable. The changes with secondary oxidation products (Barriuso and others 2013). A
in AV showed a strong increase, related to an extensive degrada- simultaneous interpretation of the results obtained for PV and AV
tion of unsaturated fatty acids. It should be noted that the decrease can therefore provide an accurate estimation of the oxidation level
observed for cycles 10 and 11 of pure oil was not logical and could of oils (Casal and others 2010; Barriuso and others 2013). Inter-
be due to a drift of the analytical technique. estingly, AV presented a higher increase until the fifth cycle than
Hydroperoxydes are primary oxidation products, which are not after, which was related to a constant PV after 5 heating cycles.
stable at high temperature, and are decomposed to more stable sec- Finally, no significant difference was found between the results

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Effect of essential oils on rapeseed oil . . .

Figure 10–Principal component analysis


applied to all samples analyzed between 0
and 15 heating cycles presented in the
score-plot (A) and the correlation circle
(B).

Food Chemistry

obtained for the 3 formulations, whether for PV or AV. The PV Changes in the tocopherols content. As shown in Figure 6,
and AV observed were similar to those obtained on vegetable oils the main isomers of tocopherols were α-tocopherol and
in other studies (Abdulkarim and others 2007; Casal and others γ -tocopherol, which correspond to the expected distribution of
2010; Talpur and others 2010; Aladedunye and Przybylski 2013). tocopherol isomers in rapeseed oil (Harwood and Gunstone 2007).
The observed evolutions of PV and AV are consistent with the The tocopherol contents in fresh oil were also conventional
reactions occurring during oxidation. Indeed, oxidation involved (Harwood and Gunstone 2007). Moreover, a strong decrease was
a radical mechanism and hydroperoxides were formed during the observed for all isomers, which indicated a degradation of these
propagation phase, but they were also decomposed to form sec- molecules. The α-tocopherol and γ -tocopherol contents were
ondary oxidation products, such as aldehydes (Choe and Min practically inexistent after 15 heating cycles. The decomposition
2006). In the case of this study, the amount of hydroperoxides of α-tocopherol seemed to be faster than that of γ -tocopherol.
formed and consumed seems almost equal from the fourth heat- Indeed, the number of heating cycles required to reduce original
ing cycle. levels by 50% was smaller for α-tocopherol than for γ -tocopherol.

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Effect of essential oils on rapeseed oil . . .

This observation was consistent with some previous studies oils, PCA was applied to the data set of all analyses performed
(Gordon and Kourimská 1995; Barrera-Arellano and others 2002), for the 3 different formulations from fresh oil over 15 heating cy-
but in contrast to other investigations (Matthäus 2006; Aladedunye cles. Figure 8 and 9 show the results for the first and the second
and Przybylski 2013). Finally, no significant difference was ob- principal component (PC1 and PC2).
served in the evolutions of the 3 studied formulations for α and Figure 10A shows the score-plot for PC1 and PC2, and
γ -tocopherols, in contrast with β and δ-tocopherols, for which Figure 10B shows the correlation circle (normed loadings) of the
the evolutions were significantly different. The decrease of β and physicochemical variables for the same PCs.
δ-tocopherols for pure oil was significantly different of the decrease PC1 and PC2 explained a great part of the total variance
observed for the oils enriched with EOs. (91.7%). The joint interpretation of the score-plot and correlation
Detailed analysis of volatile compounds. Lipid oxidation circle allowed to better understand the relationships between sam-
leads to the formation of a variety of volatile compounds and ples and physicochemical parameters. PC1 was mainly related to
the analysis of volatile compounds is important because they are the 4 tocopherol isomers on the one hand, and to AV, DPTG, TPC
related to the deterioration of flavor (Barriuso and others 2013). and FFA on the other hand (Figure 8B and 10B). It should be noted
As described in a previous study, HS sampling combined with that these 2 groups of parameters were negatively correlated. PC2
GC × GC-MS is the method of choice to separate and to identify was mostly related to PV. Interestingly, PC1 was related to com-
the volatile compounds of rapeseed oil, due to a complex matrix pounds degraded during oxidation (tocopherols) or to oxidation
Food Chemistry

(Sghaier and others 2015). products (aldehydes, DPTG, polar compounds, FFA) while PC2
To complete the previous observations, analyses of the headspace was associated with primary oxidation products (hydroperoxides).
of heated rapeseed oil with and without essential oils were per- Three different paths were discernible for the 3 different for-
formed by HS-SPME-GC × GC-MS to evaluate the effect mulations. PC1 separated pure oil from the other formulations.
of these essential oils on the formation of volatile compounds. Indeed, a time shift corresponding to 1 cycle was observed
Figure 7 shows the GC × GC chromatograms obtained for pure for the 2 oils with EOs, especially concerning the first cycles
oil and differences between the chromatograms of oils with EOs (dark arrow on Figure 10A). It was possible that the essential
and pure oil to easily visualize the differences. oils modified the induction period. This phenomenon was al-
As shown in Figure 7A and B, the headspace of fresh oil con- ready observed with other formulations of plant materials added
tained a very small amount of volatile compounds in comparison to rapeseed oil (Taha and others 2014). However, the differ-
with the heated oil. In other works, 110 volatile compounds were ence observed between fresh pure oil and fresh oils with EOs
identified in heated rapeseed oil, which highlighted a substantial on PC1 was expected because the initial samples were modi-
modification of the headspace of oil from the very first cycles fied with the addition of EOs. PC2 seemed more specific of
(Sghaier and others 2015). These compounds belonged to differ- the antioxidant effect of EOs (Figure 9A). For example, the
ent chemical families: alkanes, alkenes, furans, ketones, aldehy- coordinates on PC2 of cycles 3, 4, and 5 were always lower
des, acids, alcohols, and lactones (Sghaier and others 2015). The in the case of oils with EOs (orange arrow on Figure 10A).
formation of aldehydes was consistent with the evolution of AV These results indicated a faster degradation for pure oil than for the
described before. other formulations, which revealed a stabilizing effect of the EOs.
In order to characterize the compounds related to the 2 essen- Moreover, a slight difference between the oil with NEO and the
tial oils, 2 solutions of rapeseed oil spiked with NEO and CEO oil with CEO was also highlighted. The values observed for the
at 0.01% were prepared and analyzed. Differences between the oil with NEO on PC1 and PC2 were always slightly lower than
chromatograms of oils spiked with EOs and fresh pure oil were those observed for the oil with CEO, which suggested a better
calculated using the Chrom-Card software and allowed to identify stabilizing effect of NEO on rapeseed oil. According to the PCA,
the compounds of each essential oil (Figure 7C and E). However, the oil with NEO showed therefore the best heating stability.
these compounds were not detected in samples degraded by 1 These results were consistent with other works highlighting
heating cycle (Figure 7D and F) and they were not observed in the antioxidant potential of coriander and nutmeg (Wangensteen
samples degraded by more heating cycles. It should be noted that and others 2004; Su and others 2007; Divya and others 2012;
the differences observed in the subtracted chromatograms were Msaada and others 2014; Šojić and others 2015). Particularly, it was
mostly due to slight variations of retention times, which stresses already shown that these plants contained phenolic compounds
the necessity of a reliable 2D time alignment before any further and carotenoids, known for their antioxidant effect (Choe and
chemometric treatment of these data could be considered. Thus, Min 2009). The total phenolic content from methanolic extracts
the composition of the headspace of rapeseed oil did not seem to of nutmeg and coriander, expressed as mg of gallic acid equivalents
be deeply changed by the presence of the EOs. per gram of dry weight (mg GAE/g DW), were determined in
other studies and were of the order of 0.6 and 1 mg GAE/g DW,
Chemometric analysis respectively (Neffati and others 2011; Gupta and others 2013).
The effects of EOs on rapeseed oil were complex and due to
different interactions between different types of molecules. For Conclusions
this reason, it was necessary to perform complementary analyses, In the present work, 3 formulations were considered: pure re-
such as the determination of TPC, DPTG, PV, AV, FFA, and fined rapeseed oil and rapeseed oil enriched with 2 different es-
tocopherols contents. However, establishing connections between sential oils. Different complementary analyses were performed to
these different results was not easy. In the present study, the EOs evaluate the oxidative stability of these formulations with degra-
seemed to have a stabilizing effect, but no significant difference dation by heating cycles at 180 °C. The use of chemometric tools
was observed. In order to extract meaningful information from demonstrated the stabilizing effect of essential oils on rapeseed oil.
various analyses, the use of chemometric tools was very helpful. More specifically, rapeseed oil enriched with the NEO displayed
To have an overview of all results and to detect difference be- a better heat stability than with the CEO. Moreover, the study of
tween pure rapeseed oil and rapeseed oil enriched with essential the volatile compounds of the 3 formulations before heating and

1342 Journal of Food Science r Vol. 82, Nr. 6, 2017


Effect of essential oils on rapeseed oil . . .

after the first heating cycle showed no difference concerning the Hammer M, Schieberle P. 2013. Model studies on the key aroma compounds formed by an
oxidative degradation of ω-3 fatty acids initiated by either copper(II) ions or lipoxygenase. J
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The authors have no conflict of interest to declare. volatile secondary oxidation products in fish oil. Eur J Lipid Sci Technol 104:520–29.
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