Identification of Volatile Compounds in Cured Mexican Vanilla
Identification of Volatile Compounds in Cured Mexican Vanilla
Identification of Volatile Compounds in Cured Mexican Vanilla
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Original article
Abstract – Introduction. Headspace solid-phase microextraction combined with gas chromatography–mass spec-
trometry (HS–SPME–GC/MS) was applied to identify volatile compounds in cured vanilla (Vanilla planifolia G.
Jackson) beans originating from the state of Veracruz, Mexico. Materials and methods. A 6 kg batch from the 2012
production year was used in the study. Carboxen/polydimethylsiloxane (85 µm) fibre was selected because it had shown
a high capacity to extract detected volatiles. Results and discussion. A total of 81 volatile compounds were detected,
of which 77 were identified by comparing their mass spectra and retention times, as well as their Kovats retention in-
dices, with those of injected standards and/or by searching the National Institute of Standards and Technology’s Mass
Spectral Library database. Of those compounds, 21 shikimate derivatives, 14 terpenes/cadinenes, 12 furan derivatives,
6 esters, 7 acids, 4 ketones, 5 aldehydes, 4 hydrocarbons, 3 alcohols and 1 pyrrole were identified. Conclusion. A total
of 31 volatiles have already been reported as aroma-active compounds in cured beans of Vanilla planifolia and Vanilla
tahitensis. However, to our knowledge, 11 aromatic compounds found in this study have not previously been detected
in vanilla beans.
Résumé – Identification des composés volatils des gousses de vanille mexicaine séchées (Vanilla planifolia G.
Jackson) par HS-SPME–GC/MS. Introduction. La micro-extraction par surface de tête en phase solide couplée
avec chromatographie en phase gazeuse et spectrométrie de masse (HS–SPME–GC/MS) a été appliquée pour identifier
les composés volatils des gousses de vanille séchées (Vanilla planifolia G. Jackson) provenant de l’État de Veracruz,
au Mexique. Matériel et méthodes. Un lot de 6 kg issu de la production de l’année 2012 a été utilisé dans l’étude.
Une fibre en carboxy-polydiméthylsiloxane (85 µm) a été choisie en raison de sa grande capacité d’extraction des
composés volatils détectés. Résultats et discussion. Un total de 81 composés volatils ont été détectés, dont 77 ont été
identifiés en comparant leurs spectres de masse et leurs temps de rétention, ainsi que leurs indices de rétention Kovats,
avec ceux des standards injectés et/ou en recherchant dans la base de données de la bibliothèque de l’Institut National
des Références et de Technologie en Masse Spectrale. Parmi ces composés, ont été identifiés 21 dérivés shikimate, 14
terpènes/cadinènes, 12 dérivés du furanne, 6 esters, 7 acides, 4 cétones, 5 aldéhydes, 4 hydrocarbures, 3 alcools et 1
pyrrole. Conclusion. Un total de 31 volatils avaient déjà été rapportés comme composés aromatiques actifs dans les
gousses séchées de Vanilla planifolia et Vanilla tahitensis. Cependant, à notre connaissance, 11 composés aromatiques
présents dans cette étude n’avaient jamais encore été détectés dans les gousses de vanille.
Mots clés : Mexique / vanille / Vanilla planifolia / composé d’arôme / composé volatil
Corresponding author: apsilva30@hotmail.com
408 S. Hassan et al.: Fruits 71 (2016) 407–418
2.2 Preparation of standards operated in splitless mode for 3 min at 300 ◦ C. The oven tem-
perature program began with 3 min at 35 ◦ C, followed by a
Individual stock solutions of each commercial standard 6 ◦ C/min increase to 280 ◦ C, and then 5 min at 280 ◦ C. Detec-
were prepared in 15-mL screw-top amber vials (Supelco, tion was carried out by MS on the total ion current obtained by
Oakville, ON, Canada) as follows. Pre-sterilized tips for use electron impact at 70 eV. The mass range acquisition was m/z
with Rainin Instrument POSD pipettes (Oakland, CA, USA) 30–400. The best analytical conditions were determined using
were used for distributing the standards. A 10-µL amount of real cured vanilla bean samples. However, 25 standards were
each commercial standard was individually diluted in 10 mL injected to determine their Kovats retention indices. Volatiles
of methanol (Solution 1). A 1-mL amount of Solution 1 was were identified on the basis of their retention times when
then diluted ten-fold individually and as a group in methanol standards were available, as well as by means of searches of
(Solutions 2 and 3, respectively). A 10-µL amount of Solutions the 2005 version of the National Institute of Standards and
2 and 3 were separately diluted in 1-mL of NaCl solution (6 M) Technology’s Mass Spectral Library database. Otherwise, they
and then analyzed by HS–SPME–GC/MS. were identified on the basis of their linear retention indices as
well as electron ionization mass spectra from the literature or
from reference compounds. The linear retention index was cal-
2.3 Sampling and sample preparation culated using n-alkanes (C5–C20) as a reference.
extraction time of 30 min and desorption time of 3 min. Be- could contaminate the system and thus would still be detected
cause of the very high percentage of vanillin (85%) in rela- in the subsequent blank samples (the carry-over phenomenon).
tion to the total amount (surface area) of volatiles detected in Matrix preparation and sample volume/weight can also
cured vanilla beans from Papantla, Veracruz, Mexico [14], the strongly influence the adsorption of analytes onto the SPME
total number of volatiles detected was used instead of their fibre. At higher values for each of these parameters, reverse
total amount (surface area) to select the SPME fibre. Thus, diffusion of analytes could occur from the fibre to the sample,
81, 70 and 78 volatile compounds were detected when using resulting in a reduction of the fibre’s capacity to adsorb the an-
CAR/PDMS, CW/DVB and PA fibres, respectively. The ma- alytes [56]. A series of tests were carried out on the selected
jority of compounds detected using CW/DVB and PA fibres fibre in cut cured vanilla beans with and without the addition
were also detected using CAR/PDMS fibre; however, the sur- of 1 mL of MilliQ water or NaCl solution (6 M). These treat-
face areas of their related peaks were different for each fibre. In ments evaluated the influence of the addition of water or salt on
terms of sensitivity and selectivity, CAR/PDMS fibre provided the migration of the analytes from the matrix to the headspace.
better results than the others for volatile components and was Neither agitation nor the addition of MilliQ water or NaCl so-
thus selected for the present study. This finding concurred with lution (6 M) produced any improvement in percentage recov-
those of other studies on foodstuffs [51–53]. In fact, this fibre eries for the majority of volatiles (results not shown). A 1-g
showed the greatest capacity to extract chemical compounds mass of cured vanilla beans was sufficient to allow detection of
with a broad spectrum of polarities and molar masses [53]. targeted compounds; no significant increase in sensitivity was
observed when the sample weight was increased to 2 or even
3 g. The desorption time was set at 9 min, as a shorter time
3.1.3 Selection of HS–SPME–GC/MS parameters was not sufficient to completely desorb some analytes, partic-
The temperature of extraction and the time of extraction ularly vanillin. All trials conducted for method development
and desorption were reported previously to be the most sig- and validation used 1 g cut cured vanilla beans, CAR/PDMS
nificant factors in HS–SPME–GC/MS analysis of flavor com- fibre (85 µm), an extraction temperature of 40 ◦ C, an extraction
pounds. Factors modifying the matrix can also influence the time of 20 min, and a desorption time of 9 min. The detection
sensitivity of the fibre extraction. The addition of a salt such as limit (DL) was assumed to be less than or equal to three times
NaCl improves the effectiveness of the extraction by decreas- the signal/noise (S/N) ratio (DL ≤ 3 S/N). The quantification
ing the solubility of the analytes (the phenomenon of salting limit (QL) was assumed to be less than or equal to 10 times the
out) in an aqueous sample [54]. The pH can also modify the S/N (QL ≤ 10 S/N).
matrix; for example, the use of a 0.1 M phosphate buffer, with The repeatability of extraction by CAR/PDMS fibre was
a pH lower than the pKa of the acids involved, decreases the measured with six samples of cured vanilla beans (1 g cut
solubility of the acids and renders them more volatile. Finally, vanilla) under the conditions established above. Coefficients
sample agitation reduces the extraction time and generally im- of variation (CV) ranged from 2.2% to 21.9% (see table I).
proves extraction efficiency [55]. These parameters were all
considered in a previous study [43] and were taken into ac-
3.2 Identification of volatile components
count in the present one as well. Extraction temperatures of 40,
50 and 60 ◦ C and extraction times of 20, 30 and 40 min were The analysis of cured vanilla bean samples using HS–
tested in all combinations using CAR/PDMS fibres. A low ex- SPME–GC/MS detected 81 volatiles, of which 77 were iden-
traction temperature (e.g., 40 ◦ C) promotes the detection of tified. Of those compounds, 21 were shikimate derivatives, 14
less polar compounds, which come out of the column before were terpenes/cadinenes, 12 were furan derivatives, 6 were es-
10 min. In contrast, a high extraction temperature (e.g., 60 ◦ C) ters, 7 were acids, 4 were ketones, 5 were aldehydes, 4 were
promotes the detection of more polar compounds, which come hydrocarbons, 3 were alcohols and 1 was a pyrrole. A total of
out of the column after 10 min, specifically vanillin. Figure 1 31 volatiles have already been reported as aroma-active com-
shows the extraction percentages of vanillin using CAR/PDMS pounds in cured beans of V. planifolia and V. tahitensis (ta-
fibre at different temperatures and extraction times. The low- ble I). However, to our knowledge, 11 aromatic compounds
est percentage was obtained at 40 ◦ C 20 min−1 (10%), and the found in this study have not previously been detected in vanilla
highest percentage was obtained at 60 ◦ C 40 min−1 (100% of beans.
the scale).
A high percentage of vanillin saturates the detec-
tor [14, 17]. Of all the extraction conditions tested, the best 3.2.1 Shikimate derivatives (21)
were 40 ◦ C for 20 min−1 using CAR/PDMS fibre (see figure 2).
This choice was a compromise between the more volatile (less Shikimate derivatives were found to be the most abun-
polar) products that come out of the column before 10 min dant family and their total relative surface area was over
and the more polar products, especially vanillin, that come out 50%, 48% of which was related to vanillin. This fam-
after 10 min. Indeed, increases in the time and temperature ily comprised aldehydes (benzaldehyde, p-anisaldehyde, p-
of extraction will help increase the intensity of vanillin. This hydroxybenzaldehyde and vanillin), alcohols (benzyl alcohol,
increase in vanillin intensity has two drawbacks if one is in- p-cresol, guaiacol, p-creosol, p-anisyl alcohol and eugenol),
terested in detecting as many volatile components as possible: esters (salicylic acid methyl ester, (Z)-cinnamic acid methyl
less polar components that are present in vanilla in very small ester, (E)-cinnamic acid methyl ester, vanillic acid methyl es-
percentages may not be detected, and the high level of vanillin ter and benzoic acid phenylmethyl ester), ethers (veratrole,
Table I. Volatile compounds identified in vanilla beans by HS–SPME–GC/MS at 40 ◦ C 20 min−1 using CAR/PDMS fibre.
Compoundsa RSA (%)b CVc Kovats RId (Exp.) (Lit.) IDe Odor descriptorsf and references
Shikimate derivatives [21]
o-Cresol methyl etherg 0.10 13.7 (946) (983) MS, RI
Benzaldehyde 0.63 13.3 (956) (956) MS, RI, STD Sweet aromatic, spicy, bitter almond and dark, cherry-like [3]
Benzyl alcohol 0.33 17.6 (1026) (1021) MS, RI Chemical, fruity with balsamic nuances [3]
p-Cresol methyl ether 0.24 16.1 (1035) (1032) MS, RI Plastic, ether [2]
p−Cresol 0.78 5.7 (1071) (1075) MS, RI, STD Balsamic, wood, spicy, animal, phenolic [1–3]
Guaiacol 3.00 3.9 (1087) (1089) MS, RI, STD Chemical, sweet, smoky, aromatic, phenolic, medicinal [1–4]
1,2-Dimethoxybenzene (veratrole) 0.14 4.1 (1145) (1150) MS, RI Aromatic, somewhat phenolic, medicinal, slightly [3]
p-Creosol 0.50 4.6 (1189) (1202) MS, RI, STD Smoky, powerful cresylic [2, 3]
Salicylic acid methyl ester 0.91 4.8 (1193) (1190) MS, RI, STD Chalk, medicinal, phenolic, sweet, characteristic wintergreen [1, 3]
p-Anisaldehyde 0.08 7.7 (1257) (1251) MS, RI, STD Anise-like, almond, sweet, herbaceous-spicy, creamy, raspberry-like [2, 3, 5]
p-Anisyl alcohol 0.04 2.2 (1302) MS, STD Herbal, anise-like, sweet aromatic, balsamic, caramel, nutty, floral [1–3, 5]
(Z)-Cinnamic acid methyl ester 0.09 20.0 (1309) (1312) MS, RI, STD Sweet, fruity, balsamic, strawberry-like, cinnamon-like [1, 3, 4]
Eugenol 0.05 10.0 (1354) (1357) MS, RI Clove-live, spicy [3–5]
(E)-Cinnamic acid methyl ester 0.09 12.0 (1390) (1379) MS, RI Fruity, balsamic, strawberry-like, cinnamon-like [2–5]
p-Hydroxybenzaldehydeh 0.03 9.3 (1391) MS, STD Vanilla-like, biscuit [1]
Vanillin 48.02 7.1 (1406) (1393) MS, RI, STD Vanilla, sweet, creamy [1, 3–5]
Isovanillin 0.12 12.7 (1440) (1392) MS, RI Phenol, medicinal [5]
Acetovanillone 0.01 15.1 (1494) (1439) MS, RI Vanilla, sweet, honey [1, 3]
Vanillic acid methyl ester 0.15 11.6 (1515) (1526) MS, RI Sweet aromatic, spicy, slightly vanilla [3]
Vanillic acidh 0.04 20.6 (1567) (1560) MS, RI Sweet aromatic, somewhat vanilla; creamy, milky [3]
Benzoic acid phenylmethyl esterh 0.04 4.62 (1772) (1775) MS, RI Balsamic, oil, herb [6]
Terpenes/cadinenes [14]
m−Menth-1-eneg 1.87 8.2 (971) (987) MS, RI
S. Hassan et al.: Fruits 71 (2016) 407–418
p−Cymene 0.25 13.2 (1008) (1025) MS, RI, STD Fresh, solvent, citrus [8]
d-Limonene 2.48 3.5 (1014) (1030) MS, RI, STD Citrus-like, fresh [6, 8]
α−Terpinene 0.18 10.9 (1054) (1018) MS, RI, STD Lemony, citrusy [8]
α−Bergamotene 0.36 6.0 (1434) (1431) MS, RI Wood, warm, tea [6]
α−Curcumene 0.14 10.9 (1480) (1480) MS, RI Herb [6]
Calamenene 0.12 12.7 (1527) (1524) MS, RI Herb, spice [6]
α−Calacorene 0.07 8.7 (1548) (1550) MS, RI Wood [6]
Cadalene 0.02 11.5 (1680) (1675) MS, RI Herbal, savory [3]
α−Cubebeneg 0.04 7.9 (1348) (1348) MS, RI Herb, wax [6]
α−Copaene 0.29 4.7 (1378) (1377) MS, RI Wood, spice [6]
γ−Muurolene 0.04 6.0 (1478) (1475) MS, RI Herb, wood, spice [6]
α−Muurolene 0.05 8.3 (1500) (1502) MS, RI Wood [6]
β−Cadinene 0.16 3.3 (1520) (1519) MS, RI
Furan derivatives [12]
2-Ethylfuran 0.96 13.4 (698) (702) MS, RI
411
412
Table I. Contineud.
Compoundsa RSA (%)b CVc Kovats RId (Exp.) (Lit.) IDe Odor descriptorsf and references
Furfural 4.08 4.2 (835) (830) MS, RI, STD Sweet caramel-like, nutty, baked bread, almond [3]
2-Furyl methyl ketone (2-acetylfuran) 0.27 11.5 (882) (893) MS, RI Balsamic [3]
2-Butylfuran 1.01 18.9 (888) (893) MS, RI
γ−Butyrolactone 0.03 12.1 (944) (927) MS, RI, STD Caramel, sweet [6]
5-Methylfurfural 0.67 4.3 (952) (978) MS, RI, STD Almond, caramel, burnt [6]
2-Pentylfuran 7.94 4.8 (976) (980) MS, RI, STD Green bean, butter [6]
2-[(2E)-2-Pentenyl]furang 0.43 6.9 (983) (1000) MS, RI
2-[(1E)-1-Pentenyl]furang 0.25 15.9 (1042) (1048) MS, RI
6-(5-Methyl-furan-2-yl)-hexan-2-oneg 0.03 12.4 (1276) MS
Dihydro-5-pentyl-2(3H)-furanone (γ-nonanolactone) 0.05 10.0 (1361) (1360) MS, RI Creamy-fatty, coconut- and apricot-like [3, 4]
Dihydroactinidiolide 0.02 9.9 (1539) (1536) MS, RI
Esters [6]
Hexanoic acid methyl ester 0.44 15.6 (919) (884) MS, RI, STD Fruity, fresh, sweet [6]
Heptanoic acid methyl ester 0.08 10.1 (1005) (1006) MS, RI
2-Methyl-2-nonenoic acid methyl esterg 0.35 10.8 (1110) MS
Octanoic acid methyl ester 0.21 18.6 (1120) (1112) MS, RI Fruity, fatty [3]
Nonanoic acid methyl ester 0.19 10.3 (1219) (1224) MS, RI, STD Oily, fatty, slightly fruity [3]
Hexadecanoic acid methyl esterh 0.83 12.7 (1838) (1870) MS, RI
Acids [7]
2-Methylpropanoic acid (isobutyric acid) 0.12 16.7 (769) (765) MS, RI Buttery[1]
3-Methylbutanoic acid (isovaleric acid) 0.29 20.0 (858) (848) MS, RI, STD Buttery, oily, acid, cheese, unpleasant [1–5]
2-Methylbutanoic acid 0.47 18.8 (867) (863) MS, RI Cheese, fruity, animal, acid, sweaty, buttery [2–5]
Pentanoic acid (valeric acid) 0.07 16.0 (892) (913) MS, RI, STD Cheese, strongly acidic, caprylic [1, 3]
Hexanoic acid 0.75 6.0 (986) (990) MS, RI, STD
2-Heptenoic acid 0.39 11.4 (1115) MS
Nonanoic acid 0.08 10.0 (1263) (1272) MS, RI Oily, fatty, caprylic, cheesy [3]
S. Hassan et al.: Fruits 71 (2016) 407–418
Ketones [4]
Pentan-2-one 7.65 20.1 (682) (679) MS, RI Ether, fruit [6]
1-(2-Methyl-1-cyclopenten-1-yl)-ethanoneg 1.18 12.1 (850) (996) MS, RI
3-Octen-2-one 0.55 13.7 (1024) (1034) MS, RI Nut, crushed bug, fatty [6]
Hexahydrofarnesyl acetoneh 0.14 20.8 (1811) (1843) MS, RI Fat [6]
Aldehydes [5]
Propanal 0.28 18.4 (506) (506) MS, RI Solvent, pungent, plastic [6, 7]
2-Methylpropanal 0.31 19.7 (550) (554) MS, RI Green, pungent, burnt, malty [7]
Hexanal 1.74 12.6 (799) (803) MS, RI, STD Green, fruity [2]
Heptanal 0.61 10.9 (900) (904) MS, RI, STD Soap, fat, almond oily, rancid, powerful [6, 8]
Safranal 0.11 4.8 (1200) (1210) MS, RI, STD Herb, sweet [6]
Hydrocarbons [4]
2,4-Dimethylhexaneg 2.31 7.8 (797) (736)
(Z)-2-Octeneg 0.59 14.6 (803) (808) MS, RI
S. Hassan et al.: Fruits 71 (2016) 407–418 413
[1]: Pérez-Silva et al. [2, 14]: Brunschwig et al. [16]; [3] Zhang and Mueller [4, 17]: Takahashi et al. [18]; (http://www.flavornet.org [7, 65]: http://www.pherobase.com [66];
ID: identification. The reliability of the identification proposal is indicated by the following: MS: mass spectrum in agreement with the mass spectral database; RI: Kovats RI in agreement
o-cresol methyl ether and p-cresol methyl ether), one ke-
tone (acetovanillone), and one acid (vanillic acid). The ma-
jority of these shikimate derivatives have previously been
reported as aromatic compounds in the genus Vanilla [14,
16–19, 57], with the exception of o-cresol methyl ether.
Odor descriptorsf and references
MS, RI
Compounds are listed on the basis of their chemical family and Kovats retention index (RI) values.
[8]: http://www.crec.ifas.ufl.edu [67]. Descriptors in bold have been reported in the genus Vanilla.
tion factors of 1 953 125, 390 625 and 15 625 for anisaldehyde,
anisyl alcohol and anisyl acetate, respectively [19]. Brun-
schwig et al. [16] reported that Tahitian vanilla extract was
characterized by two families of descriptors (odors), ‘anise
with the literature; STD: mass spectrum and Kovats RI in agreement with standards.
17.9
13.4
21.9
12.3
12.3
18.8
13.9
19.4
CVc
4.1
0.15
0.23
0.08
0.06
0.03
0.37
0.89
Alcohols [3]
Others [5]
Unknown
Unknown
Unknown
Unknown
treatment [11].
Table I. Contineud.
g
h
a
f
414 S. Hassan et al.: Fruits 71 (2016) 407–418
Figure 1. Vanillin HS–SPME–GC/MS extraction using CAR/PDMS fibre (extraction temperature: 40, 50 or 60 ◦ C; extraction time: 20, 30 or
40 min).
Figure 2. Representative GC chromatogram of the volatiles in cured vanilla (V. planifolia G. Jackson), produced in Veracruz, Mexico. Peak
numbers: 1: propanal; 2: Propanal, 2-methyl-; 3: 2-Pentanone; 4: Furan, 2-ethyl-; 5: Hexane, 2,4-dimethyl-; 6: Hexanal; 2,4-dimethyl; 7:
1,3-Octadiene; 8: Furfural; 9: Ethanone, 1-(2-methyl-1-cyclopenten-1-yl)-; 10: Butanoic acid, 3-methyl-; 11: Butanoic acid, 2-methyl-; 12:
1-Propanone, 1-(2-furanyl)-; 13: Furan, 2-butyl-; 14: Heptanal; 15: Hexanoic acid, methyl ester; 16: benzaldehyde; 17: Furan, 2-pentyl-; 18:
D-Limonene; 19: 3-Octen-2-one; 20: Phenol, 4-methoxy-; 21: 3-Nonen-1-ol, (Z)-; 22: Octanoic acid, methyl ester; 23: Phenol, 2-methoxy-
4-methyl-; 24: Salicylic acid methyl ester; 25: Safranal; 26: Nonanoic acid, methyl ester; 27: 1H-Pyrrole-2,5-dione, 3-ethyl-4-methyl; 28:
Benzaldehyde, 4-methoxy-; 29: Cinnamic acid, methyl ester; 30: α-Cubebene; 31: Copaene; 32: Cinnamic acid, methyl ester, (E)-; 33: Vanillin.
m-menth-1-ene, all terpenes, namely monoterpenes (p- tributes: ‘fresh’, ‘citrus-like’, ‘lemony’, ‘herb’ and ‘wood’.
cymene, d-limonene and α-terpinene) and sesquiterpenes (α- Cadelene was identified in V. planifolia samples from Bourbon
bergamotene and α-curcumene), identified and analyzed in and Uganda as an aromatic compound with the descriptors
the studied pods have already been reported in previous stud- ‘herbal’ and ‘savory’, as reported by Zhang and Mueller [17].
ies [17, 36]. In addition to the above terpenes, eight cadinenes
were identified in vanilla, namely calamenene, α-calacorene, 3.2.3 Furan derivatives (12)
cadalene, α-cubebene, α-copaene, γ-muurolene, α-muurolene
and β-cadinene. α-Cubebene is reported for the first time in Twelve furans were identified, of which 2-[(2E)-2-
V. planifolia. Some compounds of this family have been iden- pentenyl] furan, 2-[(1E)-1-pentenyl] furan and 6-(5-methyl-
tified in vanilla samples with mainly the following flavor at- furan-2-yl)-hexan-2-one have not previously been reported
S. Hassan et al.: Fruits 71 (2016) 407–418 415
in vanilla. Dihydroactinidiolide is also reported here for the 3.2.5 Hydrocarbons (4) and alcohols (3)
first time in V. planifolia, as this furan has been reported only in
V. tahitensis, by Da Costa and Pantini [59]. The aromatic com- Only four hydrocarbons were identified in the present
pounds of this family reported in V. planifolia include 2-furyl study, which are reported for the first time in V. planifo-
methyl ketone, 2-butylfuran and γ-nonanolactone [17,18]. The lia: isomers (Z) and (E) of 2-octene, (3E)-1,3-Octadiene and
flavor attributes described for these compounds were ‘sweet’, 2,4-dimethylhexane. However, various aliphatic hydrocarbons
‘caramel-like’, ‘nutty’, ‘baked bread’, ‘almond’, ‘balsamic’, have already been reported in the literature [14, 17, 33, 36, 38].
‘creamy-fatty’, ‘coconut’ and ‘apricot-like’. According to the Thus, Ramaroson-Raonizafinimanana et al. [27] identified 25
technical details provided by the flavor and fragrance manufac- n-alkanes, 17 branched alkanes and 12 alkenes in three vanilla
turer Givaudan, dihydroactinidiolide will confer a ‘ripe, apri- bean species: V. fragrans, V. madagascariensis and V. tahiten-
cot, red fruit and wood’ organoleptic character. Furans such as sis. Because of the chemical nature of hydrocarbons, a non-
pyran (3-ethyl-4-methyl-1H-pyrrole-2,5-dione) can be formed polar fibre such as PDMS would be more suitable for their
by thermal degradation of sugars during pod curing, as in cof- extraction [54, 63, 64].
fee and cocoa [62]. Three aliphatic alcohols were identified, including 2,4-
dimethylpentan-3-ol, which is reported for the first time in
vanilla. Few volatiles belonging to this family were extracted
3.2.4 Esters (6), acids (7) and other compounds in comparison with the other groups, perhaps because of
the type of fibre used (CAR/PDMS). According to Kataoka
The aliphatic esters group reported in this study was the
et al. [54], PA fibre is more effective for alcohols and phe-
fourth most abundant family of compounds identified in the
nols. Indeed, in the preliminary trials for fibre selection (data
analyzed samples; the 2-methyl-2-nonenoic acid methyl ester
not shown), PA fibre also allowed the identification of volatile
is reported for the first time. In a study by Sostaric et al. [25]
phenols listed in the shikimate derivatives section (vanillin and
of alcoholic vanilla extract, PA fibre, which is more polar, gave
guaiacol, among others) as well as alcohols (benzyl alcohol
better results than PDMS and CW/DVB for the extraction of
and p-anisyl alcohol). However, mixed-coating fibres increase
vanillin and the esters ethyl benzoate, ethyl octanoate, ethyl
holding capacity because each coating has the effect of enhanc-
nonanoate and methyl 3-phenyl-2-propenoate (cinnamic acid
ing adsorption and distribution in the stationary phase; hence
methyl ester). Cinnamic acid methyl ester was also identified
PDMS/DVB and CAR/DVB fibres can be used for the extrac-
in this study and was grouped in the shikimate derivatives sec-
tion of more polar volatiles with low molecular weight [54].
tion along with seven other esters. Zhang and Mueller [17]
Because of the sensitivity and selectivity of CAR/PDMS fibre,
identified octanoic acid methyl ester (‘fruity’, ‘fatty’) and
it was selected for the analysis of the studied vanilla samples
nonanoic acid methyl ester (‘oily’, ‘fatty’, ‘winey’, ‘slightly
to extract shikimate derivatives, which include some phenols
fruity’) as aromatic compounds in V. planifolia extracts.
and alcohols [63].
The acids grouped in this family corresponded to aliphatic
acids with three to nine carbons and have been previously
reported in V. planifolia and V. tahitensis. Among the aro-
matic compounds reported were 2-methylpropanoic acid, 3- 4 Conclusion
methylbutanoic acid, 2-methylbutanoic acid, pentanoic acid
and nonanoic acid, with mainly attributes such as ‘buttery, A headspace solid-phase microextraction technique com-
cheese, oily and fatty’ [14, 16–19]. Studies on the evaluation bined with gas chromatography-mass spectrometry was suc-
of volatiles in coffee showed that CAR/PDMS fibre extracts cessfully applied to detect 81 volatiles, of which 77 were
more aliphatic acids with four and five carbons, whereas acetic identified, in cured vanilla (Vanilla planifolia G. Jackson)
acid and propionic acid were extracted better with CW/DVB beans originating from the state of Veracruz, Mexico. Of
fibre [63]. those compounds, 21 were shikimate derivatives, 14 were ter-
Among the four ketones identified, 1-(2-methyl-1- penes/cadinenes, 12 were furan derivatives, 6 were esters, 7
cyclopenten-1-yl)-ethanone is reported for the first time in V. were acids, 4 were ketones, 5 were aldehydes, 4 were hy-
planifolia. Hexahydrofarnesyl acetone could be detected only drocarbons, 3 were alcohols and 1 was a pyrrole. A total of
at 60 ◦ C for 20 min. 31 volatiles have already been reported as aroma-active com-
Four aliphatic aldehydes (propanal, 2-methlypropanal, pounds in cured beans of V. planifolia and V. tahitensis. How-
hexanal and heptanal) and one cyclic aldehyde (safranal) were ever, to our knowledge, 11 of these aromatic compounds have
identified. Although all these compounds have already been not previously been detected in vanilla beans. The proposed
reported in vanilla, only hexanal was reported as an aromatic semi-quantitative HS–SPME–GC/MS method has the poten-
in V. tahitensis, with the generation of ‘green and fruity’ at- tial to be used in routine analysis of volatiles in vanilla beans,
tributes [16]. The poor levels of these compounds may be due in order to evaluate rapidly the different profiles that may have
to the extraction competition between them and vanillin. In been produced in cured vanilla beans depending on their origin
fact, they all have similar chemistry, and vanillin is present at and curing process. This method allows detection of the ma-
high concentrations (1.4%), in comparison with the very low jor volatile compounds which have already been recognized in
concentrations of these aldehydes in vanilla. Studying the aro- previous work as aromatic compounds.
matic profile of vanilla pods from organic solvent extraction
allowed a greater number of compounds of this family to be Acknowledgements. This project was funded by the Instituto Tec-
identified [14, 16, 17, 38]. nológico de Tuxtepec in Mexico. The authors are grateful to the
416 S. Hassan et al.: Fruits 71 (2016) 407–418
Sistema Nacional de Recursos Fitogenéticos (SINAREFI-SNICS- Challenges and Opportunities, Perfum. Flavor. 34 (2009) 20–
SAGARPA) for financial support, and to Desarrollo Agroindustrial 22.
Gaya, S.A. de C.V. for providing vanilla bean samples, which made [9] Walton N.J., Mayer M.J., Narbad A., Vanillin, Phytochemistry
this research possible. They also wish to thank Jacinthe Fortin sin- 63 (2003) 505–515.
cerely for her valuable discussions and advice concerning this project.
[10] Shyamala B.N., Naidu M., Sulochanamma G., Srinivas P.,
Studies on the antioxidant activities of natural vanilla extract
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(Vanilla planifolia G. Jackson) originarias del estado de Veracruz, antimicrobial properties, Food Chem. 114 (2009) 791–797.
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furanos, 6 ésteres, 7 ácidos, 4 cetonas, 5 aldehídos, 4 hidrocarburos, resentative organic aroma extract from cured vanilla (Vanilla
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Cite this article as: Sabik Hassan, Pérez-Silva Araceli, Bélanger Denis, Vivar-Vera María de los Ángeles, Nicolás-García Mayra, Reyes-
López Delfino. Identification of volatile compounds in cured Mexican vanilla (Vanilla planifolia G. Jackson) beans using headspace solid-
phase microextraction with gas chromatography-mass spectrometry. Fruits 71 (2016) 407–418.