Determination of Volatile Compounds

in Apple Pomace by Stir Bar Sorptive

Extraction and Gas Chromatography-Mass
Spectrometry (SBSE-GC-MS)
C: Food Chemistry

Roberto Rodrı́guez Madrera and Belén Suárez Valles

Abstract: A method based on stir bar sorptive extraction combined with gas chromatography-mass spectrometry de-
tection (SBSE-GC-MS) has been optimized with the aim of applying it to the analysis of apple pomace. The method
allowed the identification of 124 volatile compounds after 3 h of extraction with a precision (RSDs) ranging between
2% (linalool) and 11% (ethyl hexanoate). Its use in analyzing varietal apple pomace revealed the interest of this substrate
as regards its content in aromas. From a semi-quantitative point of view, the higher content in aldehydes and esters
of the Blanquina variety is worth highlighting, as are the greater concentration of acids in the Clara variety and the
higher content of terpenes and norisoprenoides in the Coloradona variety. In contrast, the Ernestina and Perico varieties
presented the lowest levels of aromas.
Keywords: apple, aroma, GC-MS, pomace, SBSE

Practical Application: The analysis of varietal apple pomaces showed the importance of this type of waste in the food
industry, both for its content in aromas such as for its use as substrate in biotechnological processes.

Introduction (Regulation [EC] nr 110/2008 of the European Parliament of the

Cider-making is one of the main agrifood industries in As-
turias (northern Spain) with an annual average production of over
50 million liters, its principal residue being pomace (the mass re-
sulting from the pressing of the apple to obtain juice). Although
the amount of the extracted juice varies in relation to several fac-
tors such as apple variety, state of ripeness, type of pressing or
the application of different enzymatic treatments, a must yield of
around 65% to 70% (v/m) is usually estimated. This results in a
bulky residue with a great potential for application. A number
of different uses for this residue have been proposed: in the syn-
thesis of pectolytic enzymes (Berovic and Ostroversnik 1997) as
a substrate for the production of fructofuranosidase (Hang and
Woodams 1995), as an important source of antioxidant polyphe-
nols (Diñeiro Garcı́a and others 2009) or in the extraction of pectin
(May 1990), this last use being the only industrial application to
date (Gullón and others 2007).
Moreover, one of the usual agrifood-processing uses of fruit
pomaces is the production of spirits according to traditional
methods, which gives rise to quality products of recognized
prestige. Fruit marc spirits have a high alcoholic strength
and are characterized by notable levels of aroma compounds
recovered during the distillation of the fermented pomace
MS 20110659 Submitted 5/26/2011, Accepted 8/11/2011. Authors are with
Área de Tecnologı́a de los Alimentos, Servicio Regional de Investigación y Desarrollo
Agroalimentario (SERIDA), 33300-Villaviciosa, Asturias, Spain. Direct inquiries to for analysis by means of chromatographic techniques. SPME has
author Rodrı́guez Madrera (E-mail: rrodriguez@serida.org).
Council).
On the other hand, the flavors used in the agrifood industry
represent around 25% of the food additives market (Longo and
Sanromán 2006). In this respect, apple pomace could be an inter-
esting product for the sector on account of its content in aromas. In
consequence, a method for producing aromas via biotechnological
procedures has been proposed (Almonsino and others 1996).
The volatile fraction of fruits is formed by compounds belong-
ing to different chemical families such as alcohols, acids, esters,
carbonylic compounds, or terpenes, among others. Various ana-
lytical methods are used to study aromas in solid matrices of this
kind, generally consisting of a stage of extraction or concentration
of the aromas followed by a gas chromatographic analysis. Thus,
classic methods such as extraction-distillation, the principal dis-
advantages of which are the use of organic solvents and tedious
sample preparations with possible formation of several artifacts,
have sometimes been used (Ruberto and others 2008). In other
cases, methods based on headspace analysis (dynamic or static) and
purge and trap, whose principal disadvantage is the limitation of
use to the most volatile compounds, have been proposed (Rowan
and others 2009).
One of the most successful techniques for analyzing aromas in
foods has been solid phase microextraction (SPME). SPME em-
ploys a fused silica fiber coated with one or more polymers capable
of retaining the analytes of interest by absorption and/or adsorp-
tion. Subsequently, the retained aromas are thermally desorbed
several advantages, such as little or no sample preparation, the ab-
sence of organic solvents and good reproducibility. Nevertheless,

C 2011 Institute of Food Technologists

 R

C1326 Journal of Food Science r Vol. 76, Nr. 9, 2011 doi: 10.1111/j.1750-3841.2011.02406.x
Further reproduction without permission is prohibited
Aromas in apple pomace by SBSE-GC-MS . . .

the limited amount of stationary phase on the fused silica fiber
means it is not sensitive enough on occasions (Kataoka and others
2000).
Baltussen and others (1999) introduced stir bar sorption extrac-
tion (SBSE), a variation on SPME consisting of the use of magne-
tized bars covered with an absorbent polymer. SBSE presents the
same advantages as SPME, but in addition its sensibility increases
splitless mode raising the temperature from –40 to 320 ◦ C at a
rate of 10 ◦ C/s. The gas chromatograph was an Agilent 7890A
equipped with an MSD 5975C (Agilent Technologies, Palo Alto,
Calif., U.S.A.). The column employed was a FFAP (50 m × 0.2
mm i.d., 0.25 μm, Agilent Technologies). The oven temperature
program was as follows: 40 ◦ C (5 min) rising to 220 ◦ C (25 min)
at a rate of 3 ◦ C/min. Helium was used as carrier gas at a flow rate

C: Food Chemistry
around 100-fold because it uses a greater amount of stationary of 1 mL/min.
phase. SBSE has been successfully applied in recent years in the Identification was carried out by comparing spectra with those
analysis of aromas in different matrices of vegetable origin (Bicchi registered in a Wiley 138K mass spectral library and linear retention
and others 2003; Salinas and others 2004; Malowicki and others indices and confirmed by pure standards, when available.
2008). Analyses were carried out in SIM mode and the results were ex-
The aim of the present study was to develop an analytical pressed as microgram of internal standard (2-octanol) per kilogram
method based on SBSE followed by gas chromatographic anal- of wet pomace.
ysis and mass spectrometry (GC-MS) and to apply it to the study
of the aromas in apple pomace. Statistical analysis
The influence of the factors studied in the optimization of the
Material and Methods extraction method (stirring time and ionic strength) on the recov-
ery of aromas was evaluated by means of a t-test for independent
Standards
samples. The program used was SPSS version 15.0.
All standards were of analytical quality, with at least 98% purity.
The standard working solutions were prepared by dilution of com-
pounds in water. Ultra pure water was obtained from a Milli-Q Results and Discussion
system from Millipore (Mildford, Conn., U.S.A.). Optimization of the extraction method
The efficiency of the extraction by SBSE is affected by different
Samples variables: extraction time, stirring speed, and temperature, addition
A sample of multi-varietal apple pomace from an industrial cellar of inert salts or other modifiers such as methanol, pH, quantity,
and 5 mono-varietal apple pomaces from cider apples belonging and dilution of the sample, among others (Prieto and others 2010).
to different cider apple categories: Blanquina (acid), Clara (bitter), In addition, to obtain effective, reproducible extraction with stir
Coloradona (sweet-bitter), Ernestina (sweet), and Perico (semi-
acid), harvested in the year 2007, were analyzed.
To facilitate conservation, all the pomaces (composed of skin,
pulp and seeds) were dried until constant weight at 60 ◦ C for Dry pomace

48 h. Pomace moisture content ranged between 70% and 76%.

Rehydration
Pomace-Water (1:3)
Aroma extraction
Figure 1 shows the process used to obtain the extract described 30 min
subsequently.
At the time of carrying out the analysis, dried pomace was
rehydrated and mixed with double its amount of water to ob- Wet pomace
tain so-called prehomogenized pomace. An aliquot of this sample
(4.5 g) was then transferred to a 50 mL vial containing 50 mg of
Pomace-Water (1:2)
ascorbic acid and 2.8 ng of 2-octanol (internal standard, IS) and
30 mL of water. The mixture was homogenized in a Polytron PT Crushed 3 min
10–35 (Kinematica AG, Littau, Switzerland) during 2 min. Then
the polydimethylsiloxane coated stir bar (0.5 mm film thickness,
20 mm length, Twister, Gerstel GmbH & Co., Mülheim an der Pre-homogenized pomace
Ruhr, Germany) was added and the extraction was performed at
room temperature, stirring at 700 rpm for 3 h.
After the extraction, the stir bars were rinsed with distilled water, Homogenization
dried with a cellulose tissue and then placed in desorption tubes. Ascorbic acid Pomace-Water (3:20) I.S.
All analyses were carried out in triplicate.
2 min

Chromatographic analysis
Stir bar
The desorption tubes were placed in a thermal desorption unit
(TDU, Gerstel GmbH & Co.) and the stir bars were desorbed Extraction
from 25 ◦ C (1 min) to 295 ◦ C (10 min) at a rate of 60 ◦ C/min (3h)
in solvent vent mode (helium flow: 50 mL/min). The desorbed
aromas were cryofocussed in a CIS-4 programmable temperature
vaporizing (PTV) inlet (Gerstel GmbH & Co.) at –40 ◦ C using Chromatographic analysis
liquid nitrogen, the inlet liner being packed with quartz wool.
Analytes were transferred into the chromatographic column in Figure 1–Extraction procedure.

Vol. 76, Nr. 9, 2011 r Journal of Food Science C1327

 Aromas in apple pomace by SBSE-GC-MS . . .
bars, the sample mixture must be well homogenized and in liquid
form, which favors the distribution of the analytes between the
matrix and the stationary phase.
The fact that the sample is made up of skin, pulp, and seeds,
which have dissimilar compositions, means that the pomace must
be properly homogenized to guarantee adequate sampling. For
this reason, subsequent to prior testing, it was decided to add
C: Food Chemistry
double the amount of water to the wet pomace followed by a
first crushing, thereby obtaining a homogeneous, compact puree
(prehomogenized pomace), thus favoring the subsequent handling
of the sample (Figure 1). Water was then added to aliquots of
prehomogenized pomace and the particle size was reduced in
a Polytron PT 10–35. This favored the suitable transfer of the
analytes to the aqueous phase for their subsequent extraction.
Furthermore, the stirring of the bar was uniform, thus increasing
extraction reproducibility. This prior treatment was found to be
indispensable to obtain homogeneous samples and prefixed the
level of dilution and the amount of sample (pomace : water, 3 : 20).
On the other hand, desorption of the analytes extracted with
the stir bar should be complete to ensure good reproducibility of
the method and to avoid the existence of a memory effect in suc-
cessive extractions. Taking into account the compounds present in
the sample and the limitations of use of stir bars, a maximum des-
orption temperature of 295 ◦ C during 10 min was chosen. When
the stir bars were desorbed a second time under these conditions,
the chromatogram only showed peaks due to the polydimethyl-
siloxane (PDMS) film. However, as no volatiles from the pomace
were detected, this allowed us to discard any memory effect and
avoid preconditioning of the stir bar between successive extrac-
tions.
Using stir bars causes their physical degradation, with losses of
PDMS, and the deposition of non-volatile substances. This dimin-
ishes the extraction capacity and leads to a lack of reproducibility
between bars depending on the number of extractions that have
been carried out. To avoid this drawback, it was decided to use an
internal standard. 2-Octanol was thus selected taking into account
its absence in the samples and the PDMS-water distribution co-
efficient (log KO/W = 2.72), which favors an effective extraction
(David and others 2003). The extraction of this compound under
the final conditions was good, with a reproducibility of 5%, while
the relative areas to the internal standard compensated the loss of
reproducibility when the bars began to deteriorate, thus extending
their useful life.
Bearing in mind previous studies based on SBSE in similar
matrices, it was decided to perform the extractions at room tem-
perature (22 ± 2 ◦ C) at a stirring speed of 700 rpm (Bicchi and
others 2003; Salinas and others 2004).
The influence of the other parameters studied (stirring time and
ionic strength) was determined by comparing the relative areas to
the internal standard for the chromatographic peaks. To avoid in-
terferences in the estimation of the peak areas, SIM mode was used,
employing characteristic m/z relationships for each compound.
To evaluate the influence of stirring time, relative areas from
2 extraction times (1 and 3 h) were compared for 16 analytes:
ethyl hexanoate, methyl octanoate, ethyl decanoate, 2-phenylethyl
acetate, amyl alcohols, 1-octanol, 2-phenylethanol, linalool,
farnesol (isomer 2), α-farnesene, β-ionone, acetophenone,
6-methyl-5-heptenone, γ -decalactone, octanoic acid, and de-
canoic acid, covering the entire volatile range and the distinct
chemical families (Figure 2a and 2b).
The results showed different behaviors for the analytes under
study. A significant increase in extraction (P < 0.05) was detected
C1328 Journal of Food Science r Vol. 76, Nr. 9, 2011
after 3 h for ethyl decanoate, 1-octanol, farnesol, α-farnesene,
linalool, and β-ionone, while a significant decrease (P < 0.05)
in extraction efficacy was only detected for ethyl hexanoate and
2-phenylethyl acetate. No significant differences (P > 0.05) were
found for the rest of the aromas studied as regards extraction
time. It was also noted that a longer extraction time (3 h) favored
equilibrium in the distribution of analytes between the 2 phases
and improved extraction reproducibility. Under these conditions,
the reproducibility of the method ranged between 2% (linalool)
and 11% (ethyl hexanoate), values similar to those obtained with
SBSE in other matrices (Salinas and others 2004; Gomes Zuin and
others 2005).
Subsequently, the influence of ionic strength on extraction ef-
ficacy was evaluated by adding 3 g NaCl and stirring for 3 h. In
this case, a significant increase was only detected in the extrac-
tion (P < 0.05) of ethyl hexanoate, methyl octanoate, linalool,
and 2-phenylethyl acetate, which could be due to a reduction
in their solubility in the aqueous phase (Giordano and others
2009). In contrast, a significant decrease in extraction (P < 0.05)
was detected for α-farnesene, farnesol, β-ionone, 1-octanol, amyl
alcohols, γ -decalactone, and ethyl decanoate, which could have
different causes: the occupation of the surface of the PDMS by salt
ions (Portugal and others 2008) or an “oil effect,” via which the
presence of a salt in solution promotes the movement of non-polar
compounds to the surface of the aqueous phase, thus minimizing
their interaction with the PDMS (Gomes Zuin and others 2005).
As can be seen, the use of NaCl did not improve the extraction of
the analytes or the detection of new ones, so it was decided not
to use NaCl.
The analytical method allowed us to detect and identify
124 volatile compounds in apple pomace samples (Table 1), be-
longing to different chemical families, and found in the aroma of
apples by other methods (Young and others 2004; López and oth-
ers 2007; Xiaobo and Jiewen 2008; Reis and others 2009; Rowan
and others 2009).
Taking into account our interest in applying the analytical
method to study the volatile compounds in apple pomace before
and after alcoholic fermentation, known amounts of 9 aroma com-
ponents (1-octanol, linalool, methyl octanoate, ethyl decanoate,
acetophenone, decanoic acid, γ -decalactone, 2-phenylethyl ac-
etate, and 6-methyl-5-hepten-2-one), representing the different
chemical families, were added at 2 levels of addition (Table 2). The
standard additions were extracted under the optimized conditions.
The results for both matrices showed no significant differences
(P > 0.05) between the slopes of the regression lines, with a suit-
able linearity (R2 > 0.99). Thus, systematic errors such as the
matrix effect may be discarded.
Apple pomace analysis
The optimized method was applied to the analysis of 5 apple
pomaces, all derived from mono-varietal mixtures of cider apples.
Figure 3 shows the chromatogram obtained for the Perico vari-
ety. Semi-quantitative analysis was used for determination of the
volatile compounds in apple pomace. Concentrations were calcu-
lated as follows: P eISa k area
ar ea
× IS concentration. Total of 33 relevant
compounds from an odorant and technological standpoint from
different chemical families were semi-quantified (Table 3).
As can be seen, 10 carbonylic compounds were semi-quantified.
Worth highlighting among these because of their organolep-
tic relevance are the aliphatic aldehydes (2-heptenal, 2-octenal,
2-nonenal, and 2.4-decadienal isomers), responsible for herba-
ceous odors as observed in grape pomace distillates by Williams
Aromas in apple pomace by SBSE-GC-MS . . .

and Strauss (1978). The presence of unbranched chain aldehy-
des and ketones is associated with the action of different enzymes
on apple polyunsaturated fatty acids, mainly linoleic and linolenic
acids (Fauconnier and Marlier 1997). However, the most abun-
dant carbonylic compound in all cases was benzaldehyde, with a
strong smell of bitter almonds, derived from the hydrolysis of the
amygdalin present in the seeds that form part of the apple pomace.
highest concentrations of these acids were detected in the Clara
variety.
Another important group of aroma compounds are esters,
whose presence in apples is due to the esterification of the cor-
responding alcohols and carboxylic acids through the action of
an alcohol acyl-CoA transferase (Fellman and others 2000). Esters
are usually associated with fruity odors and noticeable differences

Free fatty acids are considered as the main precursors of es-
ters, alcohols and volatile aldehydes in apple (Fellman and others
2000). Octanoic, nonanoic, decanoic, and dodecanoic acids were
detected in all the analyzed samples, decanoic acid being the acid
extracted in greater amounts in all cases. On the other hand, the
C: Food Chemistry
between pomaces were observed in this study. The pomaces of
the Blanquina variety were found to present a major abundance
of esters in general, except for ethyl butyrate, more abundant in
the Coloradona variety (Table 3). Furthermore, all the samples
showed the presence of γ -decalactone, a compound responsible

Figure 2–(A) Influence of time and ionic strength on the extraction of aromas in apple pomace (mean and standard deviation). (B) Influence of time
and ionic strength on the extraction of aromas in apple pomace (mean and standard deviation).

Vol. 76, Nr. 9, 2011 r Journal of Food Science C1329

 Aromas in apple pomace by SBSE-GC-MS . . .

Table 1–Volatile compounds detected in apple pomace by SBSE-GC-MS.

Retention time Linear retention index Identification Descriptor
Alcohols
Amyl alcohols 22.53 1145 1.2 malt burnt
1-Hexanol 29.37 1276 1.2 herbaceous
1-Octen-3-ol 33.65 1372 1.2 mushroom
C: Food Chemistry

1-Octanol 38.21 1482 1.2 oily fatty

Z-2-octen-1-ol 40.80 1516 1.2 green
1-Decanol 46.19 1683 1.2 fat
Phenylmethanol 50.60 1806 1.2 floral
2-Phenylethanol 51.66 1834 1.2 rose floral
Aldehydes and ketones
Hexanal 16.99 1043 1.2 grass
Heptanal 22.19 1139 1.2 fat citrus
2-Hexenal 23.68 1165 1.2 green. apple
Octanal 27.18 1231 1 green fat
1-Hydroxy-2-propanone 27.34 1234 1.2 cheese
1-Octen-3-one 27.65 1240 1.2 mushroom
2-Heptenal 28.79 1264 1 almond fruity
6-Methyl-5-hepten-2-one 29.21 1272 1.2 green fat
2,6-Dimethyl-5-heptenal 30.12 1291 1 melon green
3-Methyl-2-cyclopenten-1-one 30.80 1306 1 fruity
2-Nonanone 31.80 1329 1.2 green
Nonanal 31.9 1333 1.2 green fat
2-Octenal 33.55 1368 1 green walnut
2-Decanone 36.00 1428 1 orange
(E,E)-2,4-heptadienal 36.22 1433 1 nuts fat
Decanal 36.64 1444 1 orange peel tallow
3,5-Octadien-2-one 37.50 1465 1 fruity mushroom
Benzaldehyde 37.54 1466 1.2 almonds
2-Nonenal 38.06 1478 1 cucumber fat
(E,E)-2,4-octadienal 40.16 1527 1 cucumber
6-Methyl-3,5-heptadien-2-one 40.26 1530 1 cinnamon coconut
2-Undecanone 40.65 1538 1.2 orange green
(Z)-2-decenal 42.43 1578 1.2 tallow
Acetophenone 42.62 1582 1.2 floral almonds
(E,E)-2,4-nonadienal 44.53 1633 1 green fat
2-Undecenal 46.49 1692 1 sweet pungent
(E,Z)-2,4-decadienal 46.85 1703 1 fried
(E,E)-2,4-decadienal 48.46 1747 1 fried
3-Phenylpropenal 56.49 1968 1.2 -
Benzophenone 70.45 2403 1 balsamic
Acids
Acetic acid 34.10 1382 1.2 vinegar
Propanoic acid 37.28 1459 1.2 pungent rancid
Crotonic acid 43.65 1607 1 burnt
Hexanoic acid 49.00 1762 1.2 sweat
Octanoic acid 56.18 1959 1.2 sweat cheese
Nonanoic acid 59.54 2063 1.2 green fat
Decanoic acid 62.77 2165 1.2 rancid fat
Benzoic acid 67.96 2331 1.2 urine
Dodecanoic acid 69.44 2374 1.2 fat
Terpenes and norisoprenoids
Linalool 37.80 1472 1.2 floral lavender
β-cyclocitral 41.64 1560 1 mint
Safranal 42.70 1526 1 herbaceous sweet
β-farnesene 43.54 1604 1 citric sweet wood
β-citral isomer 1 43.75 1610 1 lemon
β-citral isomer 2 45.58 1665 1 lemon
Camphene 45.83 1672 1 camphor
α-farnesene 46.61 1696 1 wood sweet
β-damascenone 48.80 1757 1.2 apple roses honey
Nerylacetone 49.99 1790 1 floral
Nerolidol isomer 1 51.46 1829 1 wood floral
6,10-Dimethyl-5,9-undecadien-2-ol 52.98 1869 1 fruity sweet
β-ionone 53.13 1873 1 violet raspberry
Nerolidol isomer 2 55.83 1949 1 wood floral
Pseudoionone isomer 1 56.43 1967 1 -
Pseudoionone isomer 2 59.28 2054 1 -
2-Methyl-6-methylene-1,7-octadien-3-one 61.47 2124 1 -
Farnesol isomer 1 64.25 2213 1.2 floral
Farnesol isomer 2 65.52 2255 1.2 floral
(Continued)

C1330 Journal of Food Science r Vol. 76, Nr. 9, 2011

Aromas in apple pomace by SBSE-GC-MS . . .

for the peach aroma. Lactones impart characteristics fruity odors,
on account of which these compounds are of major interest to the
industry as flavorings (Albrecht and others 1992).
Some of the constituent of vegetables with major aromatic po-
tential are terpenes and norisoprenoids. From the 8 analyzed com-
pounds belonging to this group, an isomer of farnesol was the most
abundant in the apple pomace extracts, its content in the pomace
of the Coloradona variety being noteworthy. It should also be
noted that this compound not only has a relevant aroma contri-
bution (floral odor), but that it presents significant antimicrobial
activity against human and plant pathogens (Hornby and others
2001; Jabra-Rizk and others 2006). Moreover, other terpenes such

C: Food Chemistry
Table 1–Continued
Retention time Linear retention index Identification Descriptor
Esters
Isoamyl acetate 18.76 1077 1.2 banana
Isobutyl 2-methylbutanoate 21.72 1130 1 fruity
Ethyl hexanoate 24.42 1178 1.2 apple peel fruit
Hexyl acetate 26.26 1212 1.2 herbaceous
Isoamyl 2- methylbutanoate 26.75 1222 1 apple
2-Methylbutyl 2- methylbutanoate 26.93 1226 1 apple
Z-3-hexen-1-yl acetate 28.22 1252 1.2 herbaceous
Hexyl propanoate 29.71 1283 1 fruity
Methyl octanoate 31.71 1327 1.2 orange
Butyl hexanoate 32.85 1353 1 fruity
Hexyl butanoate 33.02 1357 1.2 fruity
Hexyl 2- methylbutanoate 33.26 1363 1 strawberry
Ethyl octanoate 33.87 1377 1.2 fruity fat
Isoamyl hexanoate 34.97 1403 1 fruity
Z-3-hexen-1-yl 2- methylbutanoate 35.37 1413 1 herbaceous sweet
Ethyl nonanoate 38.16 1481 1 mulberry
Methyl decanoate 40.48 1535 1 wine
Hexyl hexanoate 41.22 1551 1.2 orange peel peach
Methyl benzoate 41.45 1556 2 lettuce raisin
Ethyl decanoate 42.24 1574 1.2 grape
Diethyl succinate 43.03 1592 1.2 wine fruity
Ethyl benzoate 43.19 1595 1.2 floral camomile
Ethyl 9-decenoate 44.06 1619 1 -
Phenylmethyl acetate 45.32 1657 1.2 floral
Ethyl phenylacetate 47.39 1718 1 fruity sweet
Methyl salicylate 47.70 1726 1 mint
Methyl dodecanoate 48.38 1745 1 coconut fat
2-Phenylethyl acetate 48.60 1751 1.2 roses honey
Ethyl 2,4-decadienoate 49.67 1781 1 pear
Ethyl dodecanoate 49.70 1781 1.2 leaves
Isoamyl decanoate 50.30 1798 1 brandy coconut
Butyl benzoate 50.45 1802 1 balsamic
2-Phenylethyl propanoate 51.30 1824 1 fruity rose
Isoamyl ethyl succinate 51.30 1825 1 -
2-Phenylethyl isobutanoate 53.71 1888 1 fruity rose
2-Phenylethyl butanoate 54.06 1897 1 floral
Isoamyl phenylacetate 54.89 1921 1 rose honey
E-11,13-dimethyl-1,2-tetradecen-1-yl acetate 56.97 1982 1 -
Ethyl tetradecanoate 57.01 1983 1.2 ether
Hexyl benzoate 57.59 2000 1 woody balsamic
Ethyl cinnamate 59.17 2051 1.2 cinnamon honey
2-Phenylethyl hexanoate 60.42 2091 1 banana pineapple
Ethyl hexadecanoate 63.44 2187 1.2 fat
Ethyl 9-hexadecenoate 64.05 2206 1 -
Ethyl octadecanoate 70.19 2396 1 -
Ethyl oleate 70.65 2409 1 mild waxy
Methyl linoleate 70.88 2416 1 -
Ethyl linoleate 72.41 2461 1.2 fat
Lactones
γ -butyrolactone 41.86 1565 1.2 caramel sweet
3-Methyl- γ -butyrolactone 45.20 1654 1 -
γ –decalactone 59.73 2069 1.2 peach
γ –dodecalactone 66.65 2292 1.2 fruity sweet
Others
Pentylbenzene 33.15 1360 1 -
Furfural 34.76 1398 1.2 caramel
5-Methylfurfural 39.42 1511 1.2 burnt sugar caramel
Furfuryl alcohol 42.80 1586 1.2 burnt
Naphtalene 46.30 1687 1 tar
2-Methylbenzotiazol 72.21 2455 1 -
1 Identification by linear retention indices and/or Mass Spectrum (quality > 85).
2 Confirmed with pure standards.

Vol. 76, Nr. 9, 2011 r Journal of Food Science C1331

 Aromas in apple pomace by SBSE-GC-MS . . .

as linalool, ß-cyclocitral, and citral, and norisoprenoids such as ß-
ionone and ß-damascenone, all of which are responsible for fruity
and floral aromas, were detected in all the samples (Guillot and
others 2006).
In agreement with other studies on apple aroma (Rowan and
others 2009; López and others 2007), alcohols constituted the
group of least relevance. Although the principal pathway of the
C: Food Chemistry
different microorganisms (Fisher and others 1999) through the
action of lipoxygenases on linoleic and linolenic acids (Manzel
and Schreier 2007).
Conclusions
The proposed analytical method has been successfully used to

formation of alcohols in food products is the fermentation of
carbohydrates and amino acids, in the case of some alcohols such
as 1-hexanol, which is responsible for herbaceous aromas, their
presence in apple products is varietal (Young and others 1996).
1-hexanol, 1-octanol and 1-octen-3-ol were the most abundant
alcohols in all cases. In this respect, it is important to emphasize
that 1-octen-3-ol (intense mushroom-like odor) is considered an
off-flavor in apple juice and its presence is usually associated with
determine volatile compounds in apple pomace, allowing the de-
tection of 124 compounds, 33 of which belonging to different
chemical families were semi-quantified. The reproducibility of the
method ranged between 2% (linalool) and 11% (ethyl hexanoate).
Its use in the analysis of varietal apple pomaces revealed the interest
of this substrate as regards its content in aromas. From a qualitative
and semi-quantitative point of view, carbonylic compounds and
esters were the most important in all the cases.

Table 2–Regression lines in fermented and unfermented pomaces from standard additions.
Unfermented pomace Fermented pomace
Addition
m/z range (mg/L) slope intercept r2 slope intercept r2
1-Octanol 84 0 to 0.282 0.328 0.006 0.999 0.328 0.015 0.997
Linaool 93 0 to 0.267 0.575 0.009 0.999 0.572 0.003 1.000
Methyl octanoate 74 0 to 098 9.436 0.004 1.000 9.510 0.022 1.000
Ethyl decanoate 88 0 to 0557 2.148 0.001 1.000 2.097 0.549 0.996
2-Phenylethyl acetate 104 0 to 0352 3.815 0.072 0.997 3.790 0.042 0.999
6-Methyl-5-hepten-2-one 55 0 to 0397 0.414 0.031 0.997 0.414 0.027 0.998
Acetophenone 105 0 to 0165 0.833 0.009 0.997 0.895 0.005 1.000
γ -decalactone 85 0 to 0150 6.972 0.055 0.992 6.480 0.037 0.998
Decanoic acid 60 0 to 3731 0.150 0.263 0.994 0.152 0.980 0.990

Abundance

I.S. 6 11+13 22
16
9000000

8000000
5

7000000

6000000

5000000 7
8 25
15
4000000
28
29 17
20 21 24
3000000
2 3 30 32 23
9 14
10 31 27 33
2000000

18 19
26
1000000 4 1 12

Time--> 15.00 20.00 25.00 30.00 35.00 40.00 45.00 50.00 55.00 60.00 65.00

Figure 3–Chromatogram obtained with the selected extraction conditions for the apple pomace from the Perico variety displaying the analyzed aromas.
I.S. Internal standard (2-octanol). Peak references are in Table 3.

C1332 Journal of Food Science r Vol. 76, Nr. 9, 2011

Aromas in apple pomace by SBSE-GC-MS . . .

Table 3–Semi-quantitative concentrations of volatile compounds in varietal apple pomaces. Mean and standard deviation expressed
as microgram of 2-octanol/kg of wet pomace.
Blanquina Clara Coloradona Ernestina Perico
Id m/z (acid) (bitter) (sweet-bitter) (sweet) (semi-acid)
Alcohols
1-Hexanol 1 56 38.84(0.78) 97.87(0.81) 65.46(5.34) 102.22(6.96) 34.22(1.33)
1-Octen3–ol 2 57 462.00(22.00) 147.11(13.33) 93.32(4.20) 66.65(2.54) 185.57(17.58)

C: Food Chemistry
1-Octanol 3 84 57.83(4.01) 60.66(4.31) 24.74(1.16) 20.90(0.98) 27.00(3.88)
Aldehydes and ketones
2-Hexenal 4 83 3.35(0.16) 9.06(0.24) 7.93(0.33) 3.33(0.23) 6.29(0.41)
2-Heptenal 5 55 543.89(4.04) 165.00(8.84) 124.79(5.37) 118.33(2.63) 355.82(34.54)
2-Octenal 6 83 548.51(25.80) 331.53(3.53) 224.87(0.19) 192.55(2.41) 358.48(17.91)
2-Nonenal 7 70 156.92(0.52) 162.8(5.41) 116.81(2.30) 75.26(0.27) 103.15(2.35)
E,Z-2,4-decadienal 8 81 518.18(7.13) 297.06(20.20) 213.10(6.69) 193.73(0.77) 328.44(25.75)
E,E-2,4-decadienal 9 81 325.13(8.29) 286.08(15.22) 207.98(9.07) 177.88(1.88) 201.54(5.45)
6-Methyl-5-hetpen-2-one 10 55 66.46(0.32) 128.65(0.69) 180.26(2.61) 138.68(7.93) 59.35(4.52)
3,5-Octadien-2-one 11 95 26.92(0.12) 43.57(0.82) 28.17(1.00) 17.24(1.18) 19.85(0.08)
6-Methyl-3,5-heptadienone 12 109 52.27(0.80) 147.81(1.43) 100.39(4.02) 69.38(1.62) 24.09(0.23)
Benzaldehyde 13 106 3074.58(60.16) 2274.46(32.53) 1001.03(114.12) 1338.75(53.45) 1560.65(133.50)
Acids
Octanoic acid 14 60 243.18(9.55) 593.78(2.72) 463.21(42.34) 374.61(12.28) 207.18(17.07)
Nonanoic acid 15 60 189.57(3.20) 525.29(0.49) 224.15(11.05) 125.14(1.50) 147.65(10.56)
Decanoic acid 16 60 711.25(7.86) 2406.82(31.61) 1933.10(180.03) 1946.22(15.17) 744.44(64.09)
Dodecanoic acid 17 60 309.05(3.83) 589.17(44.65) 476.59(36.53) 460.82(6.76) 238.04(20.56)
Terpenes y norisoprenoids
Linaool 18 93 32.41(0.66) 31.71(0.89) 21.62(0.35) 9.11(1.10) 23.21(0.48)
β-cyclocitral 19 137 12.41(0.20) 11.37(0.34) 16.10(0.24) 15.28(0.31) 13.61(0.18)
citral 20 69 222.63(7.42) 172.02(15.74) 206.48(3.34) 136.21(1.42) 156.45(16.28)
β-damascenone 21 69 71.02(1.28) 27.83(1.8) 20.77(1.40) 13.95(1.26) 38.42(3.81)
nerylacetone 22 69 646.30(7.22) 471.85(13.37) 583.59(32.22) 389.28(5.89) 490.04(39.45)
β-ionone 23 177 168.64(7.18) 184.14(0.36) 231.84(6.8) 192.76(0.85) 138.21(1.10)
Pseudo-ionone 24 69 106.03(3.37) 248.86(0.06) 363.16(6.98) 255.01(0.08) 70.05(4.44)
Farnesol (isomer 2) 25 69 1795.70(129.93) 2163.03(65.19) 3663.09(367.63) 1283.62(32.80) 784.04(66.17)
Esters and lactones
Methyl octanoate 26 74 15.94(0.79) 4.99(0.38) nd nd 8.78(0.74)
Methyl decanoate 27 74 52.57(0.75) 11.54(1.53) 4.23(0.27) nd 19.77(2.47)
Ethyl octanoate 28 88 284.96(10.65) 275.10(0.71) 178.53(15.32) 129.68(9.03) 169.44(21.64)
Ethyl decanoate 29 88 1678.08(203.36) 535.18(32.85) 300.81(16.54) 200.51(11.42) 257.66(37.28)
Ethyl benzoate 30 105 375.83(3.48) 91.49(5.99) 188.37(3.62) 102.54(3.26) 69.21(3.90)
Hexyl butyrate 31 71 9.68(0.16) 34.58(0.86) 291.84(25.87) 21.93(1.07) 12.32(1.22)
2-Phenylethyl acetate 32 104 319.29(9.83) 165.35(0.39) 49.53(2.24) 283.14(8.68) 182.42(6.35)
γ -decalactone 33 85 22.23(0.12) 33.17(2.22) 28.46(1.73) 46.84(0.43) 22.99(0.13)
Id = Identification in Figure 3, m/z = ion selected in SIM mode, nd = not detected.

Acknowledgment
Financial support for this study was managed by the Natl. Inst.
of Research and Agro-Food Technology (INIA) and co-financed
with ERDF and ESF funds (project RTA2009–00113-00–00).
References
Albrecht W, Heidlas J, Schwarrz M, Tressi R. 1992. Biosynthesis and biotechnologycal pro-
duction of aliphatic γ – and δ–lactones. In: Teranishi R, Takeoka GR, Güntert M, editors.
Flavor precursors: thermal and enzymatic conversions. Washington, D.C.: American Chemical
Society. p 46–58.
Almonsino AM, Benoussan M, Belin JM. 1996. Unsaturated fatty acid bioconversion by ap-
ple pomace enzyme system: factors influencing the production of aroma compounds. Food
Chemistry 55:327–32.
Baltussen E, Sandra P, David F, Cramers C. 1999. Stir bar sorptive extraction (SBSE) a novel
extraction technique for aqueous samples: theory and principles. J Microcol 11:737–47.
Berovic M, Ostroversnik H. 1997. Production of Aspergillus niger pectolytic enzymes by solid
state bioprocessing of apple pomace. J Biotechnol 53:47–53.
Bicchi C, Cordero C, Rubiolo P, Sandra P. 2003. Stir bar sorptive extraction (SBSE) in sample
preparation from heterogeneous matrices: determination of pesticide residues in pear pulp at
ppb (ng/g) level. Eur Food Res Technol 216:449–56.
David F, Tiemport B, Sandra P. 2003. Stir-bar sorptive extraction of trace organic compounds
from aqueous matrices. LC-GC North Am 21:108–18.
Diñeiro Garcı́a Y, Suárez Valles B, Picinelli Lobo A. 2009. Phenolic and antioxidant composition
Giordano A, Fernández-Franzón M, Ruiz MJ, Font G, Picó Y. 2009. Pesticide residue deter-
mination in surface waters by stir bar sorptive extraction and liquid chromatography/tandem
mass spectrometry. Anal Bioanal Chem 393:1733–43.
Gomes Zuin V, Montero L, Bauer C, Popp P. 2005. Stir bar sorptive extraction and high-
performance liquid chromatography-fluorescence detection for the determination of poly-
cyclic aromatic hydrocarbons in Mate teas. J Chromatogr A 1091:2–10.
Guillot S, Peytavi L, Bureau S, Boulanger R, Lepoutre JP, Crouzet J, Schorr-Galindo S. 2006.
Aroma characterization of various apricot varieties using headspace-solid phase microex-
traction combined with gas chromatography-mass spectrometry and gas chromatography-
olfactometry. Food Chem 96:147–55.
Gullón B, Falqué E, Alonso JL, Parajó JC. 2007. Evaluation of apple pomace as a raw material
for alternative applications in food industries. Food Technol Biotechnol 45:426–33.
Hang YD, Woodams EE. 1995. β–Frucofuranosidase production by Aspergillus niger species from
apple pomace. Lebensm Wiss U Technol 28:340–2.
Hornby JM, Jensen EC, Lisec AD, Tasto JJ, Janhnke B, Shoemaker R, Dussault P, Nickerson
KW. 2001. Quorum sensing in the dimorphic fungus Candida albicans is mediated by farnesol.
Appl Environ Microbiol 67:2982–92.
Jabra-Rizk MA, Meiller TF, James CE, Shirtliff ME. 2006. Effect of farnesol on Staphylococcus
aureus biofilm formation and antimicrobial susceptibility. Antimicrob Agents Chemopher
50:1463–9.
Kataoka H, Lord HL, Pawliszyn J. 2000. Applications of solid-phase microextraction in food
analysis (review). J Chromatogr A 880:35–62.
Longo MA., Sanromán MA. 2006. Production of food aroma compounds: microbial and enzy-
matic methodologies. Food Technol Biotechnol 44:335–53.
López ML, Villatoro C, Fuentes T, Graell J, Lara I, Echeverrı́a G. 2007. Volatile compounds
quality parameters and consumer acceptance of ‘Pink Lady R ’ apples stored in different

of by-products from the cider industry: apple pomace. Food Chem 117:731–8.
Fauconnier ML, Marlier M. 1997. Fatty acid hydroperoxided pathways in plants: a review. Grasas
y Aceites 48:30–7.
Fellman JK, Miller, TW, Mattinson DS, Mattheis JP. 2000. Factors that influence biosynthesis
of volatile flavor compounds in apple fruits. HortScience 35:1026–33.
Fisher G, Schwalbe R, Müller M, Ostrowski R, Dott W. 1999. Species-specific production of
microbial volatile organic compounds (MVOC) by airborne fungi from a compost facility.
Chemosphere 39:795–810.
conditions. Postharv Biol Technol 43:55–66.
Malowicki SMM, Martin R, Qian MC. 2008. Volatile composition in raspberry cultivars grown
in the pacific northwest determined by stir bar sorptive extraction-gas chromatography-mass
spectrometry. J Agric Food Chem 56:4128–33.
May CD. 1990. Industrial pectins: sources production and applications. Carbohydr Polym
12:79–99.
Menzel M, Schreier P. 2007. Enzymes and flavour biotechnology. In: Berger RG, editor.
Flavours and fragrances. Berlin: Springer. p. 489–505.

Vol. 76, Nr. 9, 2011 r Journal of Food Science C1333

 Aromas in apple pomace by SBSE-GC-MS . . .
Portugal FCM, Pinto ML, Nogueira JMF. 2008. Optimization of polyurethane foams for en-
hanced stir bas sorptive extraction of triazinic herbicides in water matrices. Talanta 77:765–73.
Prieto A, Basauri O, Rodil R, Usobiaga A, Fernández LA, Etxebarria N, Zuloaga O. 2010.
Stir-bar sorptive extraction: a view on method optimisation novel applications limitations and
potential solutions. J Chromatogr A 1217:1642–67.
Reis SFAR, Rocha SM, Barros AS, Delgadillo I, Coimbra MA. 2009. Establishment of the
volatile profile of ‘Bravo de Esmolfe’ apple variety and identification of varietal markers. Food
Chem 113:513–21.
Rowan DD, Hunt MB, Domouro A, Alspach PA, Weskett R, Volz RK, Gardner SE,
C: Food Chemistry
Chagne D. 2009. Profiling fruit volatiles in the progeny of a ‘Royal Gala’ x ‘Granny Smith’
apple (malus x domestica) cross. J Agric Food Chem 57:7953–61.
Ruberto G, Renda A, Amico V, Tringali C. 2008. Volatile components of grape pomaces from
different cultivars of Sicilian Vitis vinifera L. Bioresource Technol 99:260–8.
C1334 Journal of Food Science r Vol. 76, Nr. 9, 2011
Salinas MR, Zalacaı́n A, Pardo F, Alonso GL. 2004. Stir bar sorptive applied to
volatile constituents evolution during Vitis vinifera ripening. J Agric Food Chem 52:
4821–7.
Williams PJ, Strauss CR. 1978. Spirit recovered from heap-fermented grape marc: nature origin
and removal of the off-odour. J Sci Food Agric 29:527–33.
Xiaobo Z, Jiewen Z. 2008. Comparative analysis of apple aroma by a tin-oxide gas sensor array
device and GC/MS. Food Chem 107:120–8.
Young H, Gilbert JM, Murray SH, Ball RD. 1996. Causal effects of aroma compounds on Royal
Gala apple flavours. J Sci Food Agric 71:329–36.
Young JC, Chu CLG, Lu X, Zhu Honghui. 2004. Ester variability in apple varieties as determined
by solid-phase microextraction and gas chromatography-mass spectrometry. J Agric Food
Chem 52:8086–93.