Food Sci. Biotechnol.

20(3): 775-782 (2011)

DOI 10.1007/s10068-011-0108-4

RESEARCH ARTICLE

Volatile Distribution in Garlic (Allium sativum L.) by Solid Phase

Microextraction (SPME) with Different Processing Conditions
Na Young Kim, Min Hee Park, Eun Yeong Jang, and JaeHwan Lee

Received: 25 January 2011 / Revised: 6 April 2011 / Accepted: 6 April 2011 / Published Online: 30 June 2011
KoSFoST and Springer 2011

Abstract Treatments of autoclaving, high temperature

aging (aged-black garlic), crushing, and roasting at 100,
150, and 200oC were applied to alter the volatile profiles of
garlic (Allium sativum L.). Headspace volatiles in samples
were analyzed by a solid phase microextraction (SPME)GC/MS. Total peak areas of crushed-raw garlic were the
highest and those of aged-black garlic clove were the
lowest. Crushing effects were clearly observed in raw
garlic, aged-black garlic, and roasted garlic at 200oC for 60
min. Sulfur-containing volatiles including diallyl disulfide
and diallyl trisulfide were major volatiles. Generally, peak
areas of diallyl disulfide decreased when garlic received
autoclaving and roasting treatment while diallyl trisulfide
and allyl methyl trisulfide increased during heat treatment
compared to raw garlic. Roasting at 200oC for 60 min
caused the formation of pyrazines greatly in garlic. Principal
component analysis (PCA) for the volatile profiles by
SPME-GC/MS could discriminate types of processed
garlic successfully.
Keywords: garlic, headspace volatiles, processing effect,
solid phase microextraction

Introduction
Garlic (Allium sativum L.) is one of the oldest and
referenced herbs as food and medicinal ingredients (1).
Na Young Kim, Min Hee Park, Eun Yeong Jang, JaeHwan Lee ( )
Department of Food Science and Technology, Seoul National University of
Science and Technology, Seoul 139-743, Korea
Tel: +82-2-970-6739; Fax: +82-2-971-5892
E-mail: jhlee@seoultech.ac.kr
Na Young Kim
Seoul Metropolitan Government Research Institute of Public Health and
Environment, Gwacheon, Gyeonggi 427-805, Korea

Antibiotic, anti-yeast, cardioprotective, antihypertensive,

cancer chemopreventive, and cholesterol-lowering properties
are some representative biological activities reported in
garlic (2-5). Biological activities in garlic may be associated
with sulfur-containing compounds, which are responsible
for the characteristic pungent odor of garlic and garlic oil
(6,7).
Profiles of volatile and non-volatile compounds in garlic
are reviewed by many researchers including Lanzotti (8)
and Corzo-Martnez et al. (9). Fresh garlic contains alliin
(S-3-(2-propenylsulfinyl)-L-alanine)-, a derivative of cysteine.
When fresh garlic is crushed, allinase can convert alliin to
allicin (2-propene-1-sulfinothioic acid S-2-propenyl ester),
which is associated with the characteristic odor of crushed
fresh garlic. Garlic oil extracted by steam distillation (SD)
or simultaneous distillation and extraction (SDE) mainly
consists of allyl disulfide (4,5-dithia-1,7-octadiene) and
allyl trisulfide. Major volatiles of raw and heated garlic are
reported as sulfur-containing compounds including dimethyl
disulfide, 2-propen-1-ol, methyl-2-propenyl disulfide,
dimethyl trisulfide, diallyl disulfide, methyl-2-propenyl
trisulfide, and di-2-propenyl trisulfide (8,10,11).
Due to the garlics pungent odor, various cooking
treatments have applied to enhance its sensory attributes or
nutritional and medicinal properties. Some processing
methods including cooking in soaked water, roasting,
fermentation, steaming, hydrostatic pressure treatment, or
autoclaving have been applied to modify the off-odor of
garlic (12-14). A processing method, so-called aging at
low temperature or in the presence of alcohol, has been
introduced to make aged garlic products in Asian markets
since 1950s (15,16). Aged garlic is made through soaking
sliced garlic cloves in 15-25% ethanol for several months
at ambient temperature and the extract of aged garlic also
possessed diverse bioactivity including immunomodulatory
activity and antioxidant activity (17,18). Recently, garlic

776

with black color is made through aging the garlic bulbs in

controlled moisture contents (70-80% relative humidity) at
70oC temperature. This process can be called as high
temperature aging comparing to the traditional low
temperature aging with alcohol. In this process, garlic
bulbs without removing outer skin are treated at relatively
high temperature in the absence of alcohol for 2 weeks to
1 month duration, which is shorter period than traditional
aging process of about 20 months. The color of the aged
garlic cloves from high temperature aging process turns
from white to black or dark brown. Nutritional constituents
and sensory properties in aged-black garlic change greatly
(19). Pouch types of foods made of extracts of aged-black
garlic have been introduced in the markets and this
beverages gain popularity as healthy foods among consumers
who do not like strong and pungent odor in raw garlic.
Solid phase microextraction (SPME) is a solvent free
method for extracting and concentrating headspace
volatiles and has been successfully applied to diverse types
of foods. SPME has been used to analyze volatiles from
garlic (11) and garlic oil (20). Lee et al. (11) compared SD,
SDE, solid-phase trapping solvent extraction, and SPME as
the extraction methods for garlic volatiles and suggested
SPME could be an efficient tool to analyze volatiles of
garlic. Calvo-Gmez et al. (20) analyzed headspace volatiles
of garlic oil made by hydrodistillation using 8 different
types of SPME fibers. A total of 47 volatiles were
separated depending on the types of SPME fiber.
Although diverse treatments including high temperature
aging and roasting have adapted to treat garlic, studies on
the changes of volatile profiles in garlic from different
processing condition using SPME are rare in the literature.
The objective of this study was to analyze the distribution
of headspace volatiles in garlic treated with autoclaving,
high temperature aging (aged-black garlic), crushing, and
roasting by SPME-GC with a mass selective detector
(MS).

Materials and Methods

Materials Garlic bulbs were purchased from a local
grocery market in Seoul, Korea. Teflon-coated rubber
septa, a fiber assembly holder, 75 m carboxen/
polydimethylsiloxane (CAR/PDMS) SPME fiber, and
aluminum caps were purchased from Supelco, Inc.
(Bellefonte, PA, USA). Standard volatile compounds and
n-paraffin were purchased from Sigma-Aldrich (St. Louis,
MO, USA).
Sample preparation Garlic bulbs have several layers of
white and papery coverings and 1 garlic bulb contains
several wedge-shaped cloves covered with inner layers.

Kim et al.

Four different processing methods were applied to garlic

including autoclaving, high temperature aging, crushing,
and roasting.
Autoclaving treatment: Raw garlic bulbs without
removing outer layers were autoclaved at 121oC for 15 min
using an autoclave machine (Model SJ-220A100; Sejong
Scientific Co., Ltd., Bucheon, Korea).
High temperature aging treatment: For the aged-black
garlic samples, raw garlic bulbs without removing outer
layers were put in a cooker (Model BJC-062HT; Bubang
Techron, Seoul, Korea) and stored for 14 days at the mode
of heating without opening the door. The temperature in
the cooker was maintained at 722.5oC.
Roasting treatment: Raw cloves of garlic with layers were
roasted using a coffee roaster (Model CBR-101; Genesis
Co., Ltd., Ansan, Korea) at 100, 150, and 200oC for 30 and
60 min, respectively.
Crushing treatment: Outer layers of raw garlic cloves,
autoclaved garlic cloves, aged-black garlic cloves, and
roasted garlic cloves were peeled out and then samples
were crushed using a mortar and a pestle to simulate the
effects of chewing on the headspace volatiles in the mouth.
Tested samples in this study were raw garlic clove, rawcrushed garlic, autoclaved garlic clove, autoclaved-crushed
garlic, aged-black garlic clove, aged-black-crushed garlic,
roasted garlic clove, and roasted-crushed garlic, which
were designated as GC, CG, AGC, ACG, BGC, BCG,
RGC, and RCG, respectively. Roasted garlic at 100C for
30 min expressed as RGC100/30.
Analysis of volatiles by SPME Analysis conditions of
SPME for volatile compounds were modified from CalvoGomeza et al. (20) and Lee et al. (21). Two g of each garlic
sample was put in a 10-mL bottle and air-tightly sealed
with a Teflon-coated rubber septum and an aluminum cap.
Sample bottles were placed in the dark for 1 h at room
temperature and headspace volatiles of each garlic sample
were isolated using a 75 m CAR/PDMS solid phase at
30oC for 30 min in a circulating water bath (RW-0525G;
Lab Camp, Bucheon, Korea). The isolated volatile
compounds were determined using GC/MS. Solid phase of
SPME was exposed in an injector for 3 min. All samples
from each treatment were prepared triplicate.
GC condition Analysis conditions of SPME for volatile
compounds were adapted from Lee et al. (21). Volatiles
attached in the 75 m CAR/PDMS solid phase were
separated and identified using a Hewlett-Packard 6890 GC
equipped with a 5971A mass selective detector (Agilent
Technology, Palo Alto, CA, USA) and a DB-5ms column
(30 m0.25 mm i.d., 0.25 mm film thickness, Agilent J &
W, Folsom, CA, USA). All mass spectra were obtained at
70 eV and 220oC ion source temperature. The identification

Volatiles in Processed Garlic by SPME

of compounds was made by a combination of NIST Mass

Spectra, linear retention indices (RI) of n-paraffin as
external references, and GC retention times of some
standard compounds. Helium was carrier gas at 1.0 mL/
min and the oven temperature was held at 40oC for 2 min
and increased from 40 to 160oC at 6oC/min and from 160
to 220oC at a rate of 10oC/min.
Statistical analysis Results of total peak areas and
selected major volatiles were statistically analyzed by
analysis of variance (ANOVA) and Duncans multiple
range test using commercially available software package
SPSS software program (SPSS Inc., Chicago, IL, USA). A
p value <0.05 was considered significant. Principal
component analysis (PCA) for volatiles in garlic by SPME
was conducted using covariance matrix with no rotation
and SPSS software program.

Results and Discussion

Distribution of volatiles in raw, autoclaved, aged-black,
and roasted garlic samples Changes of total ion counts
in garlic treated with different processing conditions are
shown in Fig. 1. Raw-crushed samples (CG) and agedblack garlic cloves (BGC) showed the highest and lowest
total ion counts among tested samples (Fig. 1). Agedblack-crushed garlic (BCG) had 11 times more total ion
counts than uncrushed samples (BGC). The lowest total
ion counts in BGC could be due to the presence of layers
in cloves, which may prevent the release of compounds
from the garlic cloves into the headspace. Generally, garlic
roasted at 150oC for 30 and 60 min had less total volatiles

777

compared to raw garlic and roasted garlic at 100oC. Much

variation was observed in total volatiles of garlic samples
among autoclaving, high temperature aging, roasting, and
crushing treatments. Calvo-Gomeza et al. (20) compared
commercially available 8 types of SPME solid phases for
the headspace volatiles of garlic oil and reported that solid
phase of 75 m CAR/PDMS detected the maximum
number of volatiles compared to other SPME solid phases.
Although 75 m CAR/PDMS could not detect all volatiles,
our study adapted 75 m CAR/PDMS for the volatile
analysis based on the reports of Calvo-Gomeza et al. (20).
Major volatiles in garlic with raw, autoclaving, and high
temperature aging treatments are shown in Table 1.
Number of volatiles identified in GC, CG, AGC, ACG,
BGC, and BCG were 21, 18, 21, 20, 12, and 17,
respectively. Out of 21 volatiles in GC and CG, 17 sulfurcontaining compounds consisted more than 88.5% of total
ion counts. Most volatiles in raw garlic are diallyl
disulfides (46.14% of total volatiles in GC and 40.88% of
total volatiles in CG), 3-vinyl-1,2-dithiacyclohex-5-ene,
3-vinyl-1,2-dithiacyclohex-4-ene, and diallyl trisulfide
(5.99% of total volatiles in GC and 9.32% of total volatiles
in CG). 2-Propen-1-ol and 2-methyl- 3,4-dihydro-2Hthiopyran, which were detected in GC, were not observed
in CG.
Dimethyl disulfide, which was not observed in raw
garlic samples (GC and CG), was observed in autoclaved
samples (AGC and ACG), indicating thermal reaction may
cause the formation of this compound. Yu et al. (27) and
Calvo-Gomez et al. (20) reported the detection of dimethyl
disulfide in garlic essential oil prepared from water or
solvent extraction. The boiling point of dimethyl disulfide
is 109-110oC and higher temperature may increase the

Fig. 1. Changes of total peak areas in garlic treated with different processing conditions. Different letters on the bar are significant at
p<0.05.

778

Kim et al.

Table 1. Effects of autoclaving, high temperature aging, and crushing process on the distribution of major volatiles in garlic
RI1)
574
600
697
744
856
888
909
918
944
980
989
1,010
1,098
1,104
1,160
1,176
1,183
1,191
1,201
1,220
1,250
1,328
1,355
1,432
1,535

Compound
Sulfur dioxide
2-Propen-1-ol
Allyl mercaptan
Allyl methyl sulfide
Dimethyl disulfide
Allyl sulfide(diallyl sulfide)
2-Methyl-3,4-dihydro-2H-thiopyran
3,4-Dimethylthiophene
Allyl methyl disulfide
1,3-Dithiane
Phenylethyl butyrate
Dimethyl trisulfide
2-Methyl-5-ethylpyrazine
Diallyl disulfide
1,5-Dithiocane
Allyl methyl trisulfide
1-Methyl-3-pyrrolin-2-one
O-Methyl 2-acetylhydrazinecarbothioate
1,3,5-Trithiane
2,4-Dimethylthiazole
3-Vinyl-1,2-dithiacyclohex-4-ene
3-Vinyl-1,2-dithiacyclohex-5-ene
Diallyl trisulfide
1-Methylimidazole-2-thiol
5-Methyl-1,2,3,4-tetrathia-cyclohexane
1-Hydoxy-4-methyl-2,6-di-tert-butylbenzene

Total volatiles (108 ion counts)

GC2)

CG
3)

0.550.18c
0.150.02bc
1.270.03bc
0.280.01ab
ND5)
1.500.51a
0.200.28a
0.710.07c
2.150.20d
0.620.00c
1.220.13a
ND
ND
23.502.66d
ND
0.900.13b
0.510.11b
ND
0.240.05ab
0.470.00b
5.131.78b
6.211.71c
3.050.27b
1.530.01a
0.480.04a
0.260.06b

0.480.02bc
0.090.06ab
2.100.74c
0.120.17a
ND
1.580.26a
0.340.21a
0.780.00c
1.330.10c
0.360.03b
2.790.62b
ND
ND
23.511.73d
ND
0.990.51b
0.230.01a
ND
0.550.34b
0.440.00a
6.861.97b
6.431.07c
5.361.96b
2.051.14a
1.550.84ab
ND

AGC

ACG

0.160.00a 0.130.03a
0.120.02bc 0.030.05a
1.430.50ab 1.060.02b
0.240.04ab 0.210.01ab
0.040.01a 0.030.00a
1.960.25a 1.500.24a
0.240.18a 0.290.04a
0.270.04b 0.190.04b
1.240.26c 0.820.11b
0.570.10c 0.360.05b
3.300.97b 3.590.31b
ND
ND
ND
ND
18.700.64c 13.601.29b
ND
ND
3.560.35c 2.890.42c
0.450.02b 0.310.07a
ND
ND
1.240.24c 1.070.10c
ND
ND
2.040.60a 1.490.14a
2.700.12b 2.190.33b
9.111.20c 9.610.28c
2.080.13a 1.820.06a
2.641.03b 1.960.33b
0.540.12c 0.430.11c

BGC

BCG

0.080.01a
0.010.00a
0.070.03a
0.370.02b
0.040.01a
1.850.08a
ND
0.010.00a
0.140.02a
0.020.01a
ND
0.010.00
ND
0.490.06a
ND
ND
ND
0.060.00a
ND
ND
ND
ND
0.030.00a
ND
ND
0.040.04a

0.350.02b
0.200.00b
0.530.00ab
0.860.01c
0.260.10b
5.440.10b
0.810.05a
0.080.00a
2.000.03d
ND
ND
ND
0.590.03
12.890.34b
10.400.39
3.540.12d
ND
6.540.18b
ND
ND
0.850.02a
2.500.06b
4.040.03b
ND
2.570.04b
0.180.00ab

1)

Linear retention indices (RI) were determined using n-paraffin as external references.
GC, raw garlic clove; CG, raw-crushed garlic; AGC, autoclaved garlic clove; ACG, autoclaved-crushed garlic; BGC, aged black garlic clove,
and BCG, aged-crushed black garlic
3)
MeanSD (n=3); ND, not detected; Different letters are significant at p<0.05 among the same row.
2)

volatility of dimethyl disulfide. In raw garlic, dimethyl

disulfide may not have enough high volatility to be
detected by SPME fiber. Major volatiles in AGC and ACG
are in the order of diallyl disulfides (35.53% of total
volatiles in AGC and 31.23% of total volatiles in ACG),
diallyl trisulfide (17.31% of total volatiles in AGC and
22.07% of total volatiles in ACG), and allyl methyl
trisulfide. The contents of diallyl trisulfide were the most
increased volatile in autoclaved samples compared to those
in raw samples. Diallyl trisulfide, diallyl disulfide, and allyl
methyl trisulfide were reported as major sulfur-containing
volatiles in steamed garlic for 15 min and autoclaved garlic
for 5 min (22).
Distributions and number of volatiles in BGC were
substantially different from those of BCG. Allyl sulfides
consisted 58.3% of total volatiles in BGC and 10.8% of
total volatile in BCG. 3,4-Dimethylthiophene, 2-methyl-5ethylpyrazine, and 1,5-dithiocane, which were not detected
in BGC were found in BCG. 3,4-Dimethylthiophene and 2-

methyl-5-ethylpyrazine may be already formed and present

inside in the matrix of BGC during high temperature aging
process, which were liberated into the headspace when the
structure of matrix was crushed. Due to the temperature
over 72, enzymatic reactions for the formation of color and
volatiles may be inhibited during high temperature aging
process. Dark color can be developed from non-enzymatic
reactions including Maillard browning reaction and
caramelization (19). In low temperature aging process,
diallyl sulfide, diallyl disulfide, diallyl trisulfide, and
dithiin have been found in aged garlic through the activity
of allinase. In case of conditions of low enzyme activity, Sallyl cysteine and S-allyl mercaptocysteine were main
sulfur-containing non-volatiles in aged garlic (23).
Major volatiles identified from roasted garlic samples
are shown in Table 2. The number of volatiles in roasted
garlic samples was greatly influenced by the roasting time,
roasting temperature, and crushing process. All the 100
roasted garlic samples, RGC150/30, and RCG150/30 had

Sulfur dioxide
2-Propen-1-ol
Allyl mercaptan
Allyl methyl sulfide
Dimethyl disulfide
3-Methyl-thiophene
2-Methylpyrazine
Allyl sulfide(diallyl sulfide)
2-Methyl- 3,4-dihydro- 2H-thiopyran
3,4-Dimethylthiophene
Allyl methyl disulfide
1,3-Dithiane
Phenylethyl butyrate
2-Methyl-3-ethylpyrazine
Diallyl disulfide
1,5-Dithiocane
Allyl methyl trisulfide
1-Methyl-3-pyrrolin-2-one
2,4-Dimethyl-6,7-dihydro-5Hcyclopenta[D]pyrimidine
1,3,5-Trithiane
2,5-Dimethyl-3-isobutylpyrazine
3-Vinyl-1,2-dithiacyclohex-4-ene
3-Vinyl-1,2-dithiacyclohex-5-ene
Diallyl trisulfide
1-Methylimidazole-2-thiol
5-Methyl-4,7-dithiadeca-1,9-diene
5-Methyl-1,2,3,4-tetrathia-cyclohexane
1-Hydoxy-4-methyl-2,6-di-tert-butylbenzene

Compound

ND

0.20.0
0.10.0
1.00.4ab
0.30.0ab
ND
ND
ND
2.70.1ab
0.40.0
0.20.0
1.20.1ab
0.20.1
2.91.0
ND
15.82.0bc
ND
2.90.4a
0.40.0
ND

ND

ND

0.20.1
0.30.0
0.10.1
0.150.02 0.10.1
0.30.0
1.00.3ab 1.10.0b 0.60.2ab
0.30.0ab 0.20.0a 0.30.1a
ND
ND
ND
ND
ND
ND
ND
ND
ND
2.80.2abc 2.40.1a 2.20.3a
0.40.1
0.20.2
0.30.0
0.20.0
0.10.0
0.10.1
0.90.6ab 0.60.1a 0.90.2ab
0.40.0
0.20.0
0.30.1
3.31.0
4.10.1
0.71.3
ND
ND
ND
16.30.8c 11.41.5abc 10.61.3ab
ND
ND
ND
3.60.6ab 2.30.1a 2.40.6a
0.40.0
0.20.0
0.30.3
ND

ND

2)

ND

ND

ND

0.20.0
0.20.0
0.20.0
0.20.0
0.30.1
0.20.0
0.50.1
0.30.1
0.30.0
0.40.1
0.50.3a 0.870.17ab 0.80.2ab 1.00.3ab 0.80.2ab
0.30.0ab 1.10.5d 0.60.1bc 0.70.0c 0.70.1c
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
0.30.3a
2.80.1abc 4.11.7cd 3.80.5bcd 4.21.1d 4.20.9d
0.40.0
0.10.1
0.10.1
0.10.1
0.10.1
0.20.0
0.10.0 0.10.01 0.10.0
0.10.0
0.90.1ab 3.71.4e 1.80.2bc 2.50.2cd 2.10.4cd
0.40.0
0.40.0
0.30.0
0.30.0
0.30.0
1.60.4
1.41.9
2.70.4
3.21.1
1.81.6
ND
ND
ND
ND
0.30.3a
13.51.2bc 12.02.8abc12.30.4abc12.52.9abc12.31.8abc
ND
ND
ND
ND
ND
2.30.4a 6.50.4cd 4.60.3abc 5.60.5bcd 3.43.0ab
0.40.0
0.60.2
0.50.0
0.50.1
0.30.3

ND

ND
0.20.1
0.80.2
2.40.6
5.40.6ab
ND
0.90.3
1.30.3
0.30.1

6.80.5

0.30.0
0.40.0
0.80.7
0.30.0
ND
0.70.1ab
1.10.0d 0.70.1e
ND
0.20.0
ND
0.20.0
0.10.0b. 0.10.0b
4.90.6d 9.20.2e
0.10.1
0.10.0
0.10.1
0.10.0
2.50.6cd 3.00.0de
ND
ND
ND
ND
0.70.1b 1.00.1c
7.81.3a 12.20.0abc
ND
2.60.1
2.20.4a 7.90.8d
ND
ND

1.50.1
0.90.3
1.20.0
1.30.0
0.70.1
0.70.1
1.00.0
1.10.1
1.10.2
1.00.2
0.60.0
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
2.60.2
2.20.5
2.70.1
1.80.1
1.00.4
1.00.3
1.10.2
1.50.3
1.40.5
1.20.3
0.40.1
3.81.3
3.40.5
3.70.1
2.50.0
1.60.5
1.60.3
2.20.3
2.40.3
2.30.6
2.10.4
1.00.1
13.40.1e 13.51.5e 12.42.6de 10.90.0cde 6.42.3abc 6.21.8abc 8.10.8bcd 6.75.4abc 9.82.3bcde 7.91.6bcd 2.60.7a
2.30.1
1.90.4
1.80.1
1.40.1
0.70.1
0.70.2
ND
0.40.3
ND
0.20.3
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
ND
2.90.1
2.00.9
2.40.1
2.50.1
1.30.4
1.30.3
2.00.2
2.20.3
2.20.6
1.90.5
0.80.2
0.80.1
0.50.1
0.50.0
0.40.1
0.40.1
0.40.1
0.80.4
0.50.1
0.60.1
0.50.1
0.40.0

ND

0.30.0
0.10.0
1.10.0b3)
0.20.0a
ND
ND
ND
2.40.4a
0.40.0
0.30.0
1.20.3ab
0.40.1
2.82.5
ND
14.95.4bc
ND
3.80.5ab
0.50.1

RGC100/ RCG100/ RGC100/ RCG100/ RGC150/ RCG150/ RGC150/ RCG150/ RGC200/ RCG200/ RGC200/ RCG200/
302)
30
60
60
30
30
60
60
30
30
60
60

Total volatiles (108 ion counts)

Linear retention indices (RI) were determined using n-paraffin as external references.
Roasted garlic clove and roasted-crushed garlic at 100oC for 30 min were expressed as RGC100/30 and RCG100/30, respectively.
3)
MeanSD (n=3); ND, not detected; Different letters are significant at p<0.05 among the same row in the selected volatiles.

1)

1,191
1,212
1,220
1,250
1,328
1,355
1,355
1,432
1,535

1,184

574
600
697
744
772
844
856
888
909
918
944
980
1,014
1,098
1,104
1,160
1,176

RI

1)

Table 2. Distribution of major volatiles detected in garlic with roasting and crushing treatments

Volatiles in Processed garlic by SPME

779

780

Kim et al.

Fig. 2. Relative contents (%) of diallyl disulfide, diallyl trisulfide, and allyl methyl trisulfides to total sulfur-containing compounds
in garlic treated with different processing conditions. Different letters are significant at p< 0.05 among the same volatiles; ND, not
detected

21 volatiles. However, the number of volatiles in RGC150/

60, RCG150/60, RGC200/30, RCG200/60, RGC200/60,
and RCG200/60 was 19, 20, 18, 20, 17, and 23,
respectively. Like other garlic samples, diallyl disulfide
was the most detected volatile. Due to the input of thermal
energy, pyrazines including 2-methylpyrazines, 2-methyl3-ethylpyrazine, and 2,5-dimethyl-3-isobutylpyrazine were
observed in garlic samples roasted at 200oC for 60 min.
However, these pyrazines were not observed in 100 and
150oC roasted garlic samples and even in RGC200/30, and
RCG200/30. The contents of diallyl trisulfide were high in
garlic roasted at 100oC and tended to decrease in samples
roasted at 150 and 200oC, which implies formation and
decomposition of diallyl trisulfide are depending on the
thermal energy. Considering the contents of diallyl
trisulfide in raw and autoclaved garlic, diallyl trisulfide
seems to be formed in garlic treated with around 100-120
o
C thermal energy but decomposed at higher temperature
like 150 and 200oC in garlic.
Maillard reaction has been regarded as major mechanisms
for the formation of pyrazines and pyridine in potato chips
(24) and in roasted sesame seed oil (25). 2-Methylpyrazine
was detected from Maillard reaction of glutathione and
reducing sugars such as glucose or fructose (26).
Six volatiles including dimethyl disulfide, 3-methylthiophene, 1,5-dithiocane, 2,4-dimethyl-6,7-dihydro-5hcyclopenta[D]pyrimidine, 2,5-dimethyl-3-isobutylpyrazine,
and 5-methyl-4,7-dithiadeca-1,9-diene were found additionally
in RCG200/60, which implies these compounds may be
located inside of garlic matrix and crushing process helps
to release these volatiles to the headspace.

Volatiles of 1,3-dithiane, phenethyl butyrate, and 1methyl-3-pyrroin-2-one, which were detected in 100 and
150oC roasted samples, were not observed in RGC200/60,
and RCG200/60. 1,3-Dithiane and phenethyl butyrate were
found in raw and autoclaved samples implying these
compounds may be heat-labile and/or be changed into
other volatile or non-volatile forms.
Peak areas of 2-propen-ol and allyl methyl sulfide
started to increased in garlic roasted at 150oC and 3-vinyl1,2-dithiacyclohex-4-ene and diallyl trisulfide were
decreased from 150oC roasted samples.
Changes of relative contents of diallyl disulfide, diallyl
trisulfide, and allyl methyl trisulfides to the total sulfurcontaining compounds in garlic with different processing
conditions are shown in Fig. 2. Diallyl disulfide is the most
abundant sulfur-containing compound in garlic ranging
from 48% in GC to 18% in BGC. Relative contents of
diallyl disulfide in raw garlic clove (GC) were significantly
higher than those of other garlic samples and diallyl
trisulfide and allyl methyl trisulfide in GC were significantly
lower than other samples (p<0.05). Application of thermal
energy changed the profiles of sulfur-containing compounds
greatly. Generally, relative contents of diallyl disulfide
decreased in aged-black and roasted garlic and those of
diallyl trisulfide and allyl methyl trisulfide increased. Allyl
methyl trisulfide was not found in BGC, which may be due
to the inhibiting effects of layers in aged-black garlic clove.
Some sulfur-containing volatiles including 3-vinyl-1,2dithiacyclohex-5-ene and 3-vinyl-1,2-dithiacyclohex-4-ene
decreased upon autoclaving or roasting process.
Number of identified volatiles in this study was relatively

Volatiles in Processed Garlic by SPME

781

small compared to that of volatiles from previous reports.

Because SPME method just isolates and concentrates the
headspace volatiles adsorbed on the solid phase, number
and types of volatiles from SPME method are limited
compared to those from solvent extraction and purge and
trap methods.
Many volatiles identified in this study were already
reported in the previous reports of garlic (4,11,26). Woo et
al. (4) heated garlic bulb at 100, 110, 120, and 130oC and
extracted the volatiles using SDE and reported that the
major volatiles of raw and heated garlic were dimethyl
disulfide, 2-propen-1-ol, allyl methyl disulfide, dimethyl
trisulfide, diallyl disulfide, allyl methyl trisulfide, and
diallyl trisulfide. Lee et al. (11) reported that allyl methyl
sulfide, allyl sulfide, diallyl disulfide, and 1,3-dithiane
were representative volatiles from garlic samples using 50/
30 m divinyl benzene/carboxen/polydimethylsiloxane
(DVB/CAR/PDMS) SPME fiber. Yu et al. (27) identified
allyl methyl trisulfide, diallyl disulfide, diallyl trisulfide,
dimethyl disulfide,1,3-dithiane, aniline, 2,4 dimethylfurna,
and 2-propen-1-ol using essential oils from raw crushed
garlic treated with diverse extraction methods including
steam distillation, water distillation, and SDE. Difference
in the number and profiles of detected volatiles may be due
to the volatile analysis and garlic processing methods.
PCA analysis of volatiles in garlic treated with diverse
processing PCA was conducted to determine the relation
among volatile distribution and garlic samples received
different processing. Loading and score plots of PCA for
volatiles in garlic samples are shown in Fig. 3. The first
principal component (PC1) and second principal component
(PC2) expressed 38.07 and 20.24% of the volatile variability
among headspace volatiles in diversely processed garlic,
respectively. Volatiles that are positively correlated to PC1
are pyrazines, allyl methyl disulfide, allyl methyl trisulfide,
and allyl sulfide and negatively correlated volatiles to PC1
are diallyl disulfide and allyl mercaptan (Fig. 3A). According
to the scoring plot, PCA clearly distinguished garlic
samples with different processing methods using volatile
profiles (Fig. 3B). Raw garlic clove and raw-crushed garlic
were grouped together and located negatively to PC1 score
while garlic roasted at 200oC for 60 min and aged-black
garlic samples were located positively. Samples autoclaved
and roasted at 100 and 150oC were grouped together (Fig.
3B). As roasting temperature increased from 100 to 200oC,
the PCA plot extended from the left side (negative value of
PC1 score) through the middle to the right side (positive
value). Loading and score plots of PCA showed that diallyl
disulfide and allyl mercaptan were more related with raw
garlic samples (CG and GC), while diallyl trisulfide and
pyrazines were more correlated with autoclaved and
roasted-crushed garlic at 200oC for 60 min, respectively.

Fig. 3. Loading (A) and score (B) plots of PCA for the
headspace volatiles in garlic treated with different processing
conditions by SPME-GC/MS.

PCA approaches using headspace volatiles are useful

techniques for differentiating foods prepared differently
(28). Park et al. (25) used PCA technique for the analysis
of headspace volatiles to discriminate sesame oil prepared
from sesame seeds roasted with different condition. PCA
using headspace volatiles can be a useful tool to
discriminate garlic prepared from different processing.
In conclusion, headspace volatiles in garlic samples
treated with autoclaving, high temperature aging, roasting,
and crushing process were analyzed by SPME-GC/MS and
volatile data were processed with PCA. Sulfur-containing
compounds including diallyl disulfide, allyl mercaptan,
allyl sulfide, and diallyl trisulfide were major volatiles in
garlic samples. Generally, diallyl disulfide decreased and
diallyl trisulfide increased upon receiving thermal energy
and pyrazines appeared in garlic roasted at 200oC for 60
min and aged-black garlic samples. PCA showed that
roasted or aged-black garlic could be differentiated from
raw garlic using profiles of headspace volatiles. This is the
first report comparing the distribution of volatiles using
SPME method from garlic and crushed garlic samples
prepared by diverse processing such as autoclaving, high

782

temperature aging, and roasting. Further studies are needed

to correlate among the changes of sensory attributes and
volatile profiles in diversely processed garlic.

Kim et al.

15.
16.

References
17.
1. Rivlin RS. Historical perspective on the use of garlic. J. Nutr. 131:
951-954 (2001)
2. Sadi AM, Toda T, Oku H, Hokama S. Dietary effects of corn oil,
oleic acid, perilla oil, and evening primrose oil on plasma and
hepatic lipid level and atherosclerosis in Japanese quail. Exp. Anim.
45: 55-62 (1996)
3. Onogi N, Okuno M, Komaki C, Moriwaki H, Kawamori T, Tanaka
T, Mori H, Muto Y. Suppressing effect of perilla oil on
azoxymethane-induced foci of colonic aberrant crypts in rats.
Carcinogenesis 17: 1291-1296 (1996)
4. Woo KS, Yoon HS, Lee YR, Lee JS, Kim DJ, Hong JT, Jeong HS.
Characteristics and antioxidative activity of volatile compounds in
heated garlic (Allium sativum). Food Sci. Biotechnol. 16: 822-827
(2007)
5. Kim JW, Choi JH, Kim YS, Kyung KH. Antiyeast potency of
heated garlic in relation to the content of allyl alcohol thermally
generated from alliin. J. Food Sci. 37: 185-189 (2006)
6. Oi Y, Okamoto M, Nitta M, Kominato Y, Nishimura S, Ariga T,
Iwai K. Alliin and volatile sulfur-containing compounds in garlic
enhance the thermogenesis by increasing norepinephrine secretion
in rats. J. Nutr. Biochem. 9: 60-66 (1998)
7. Block E. The organosulfur chemistry of the genus alliumimplications for the organic chemistry of sulfur. Angew. Chem. Int.
Edit. 31: 1135-1178 (1992)
8. Lanzotti V. The analysis of onion and garlic. J. Chromatogr. A 1112:
3-22 (2006)
9. Corzo-Martnez M, Corzo N, Villamiel M. Biological properties of
onions and garlic. Trends Food Sci. Tech. 18: 609-625 (2007)
10. Chyau CC, Mau JL. Release of volatile compounds from
microwave heating of garlic juice with 2,4-decadienals. Food Chem.
64: 531-535 (1999)
11. Lee SN, Kim NS, Lee DS. Comparative study of extraction
techniques for determination of garlic flavor components by gas
chromatography-mass spectrometry. Anal. Bioanal. Chem. 377:
749-756 (2003)
12. Chung SK, Seog HM, Choi JU. Changes in volatile sulfur
compounds of garlic (Allium sativum L.) under various drying
temperatures. Korean J. Food Sci. Technol. 26: 679-682 (1994)
13. Lim CY, Hong EJ, Noh BS, Choi WS. Effect of high hydrostatic
pressure and pH on the reduction of garlic off-flavor. Korean J.
Food Sci. Technol. 42: 533-540 (2010)
14. Sohn KH, Lim JK, Kong UY, Park JY, Noguchi A. High pressure

18.
19.
20.

21.

22.
23.
24.

25.

26.
27.
28.

inactivation of alliinase and its effects of flavor of garlic. Korean J.

Food Sci. Technol. 28: 593-599 (1996)
Imai J, Ide N, Nagae S, Moriguchi T, Matsuura H, Itakura Y.
Antioxidant and radical scavenging effects of aged garlic extract and
its constituents. Planta Med. 60: 417-420 (1994)
Moriguchi T, Saito H, Nishiyama N. Aged garlic extract prolongs
longevity and improves spatial memory deficit in senescenceaccelerated mouse. Biol. Pharm. Bull. 19: 305-307 (1996)
Dillon SA, Burmi RS, Lowe GM, Billington D, Rahman K.
Antioxidant properties of aged garlic extract: An in vitro study
incorporating human low density lipoprotein. Life Sci. 72: 15831594 (2003)
Chandrashekar PM, Venkatesh YP. Identification of the protein
components displaying immunomodulatory activity in aged garlic
extract. J. Ethnopharmacol. 124: 384-390 (2009)
Shin JH, Choi DJ, Lee SJ, Cha JY, Kim JG. Sung NJ. Changes of
physicochemical components and antioxidant activity of garlic
during its processing. J. Life Sci. 18: 1123-1131 (2008)
Calvo-Gmez O, Morales-Lpez J, Lpez MG. Solid-phase
microextraction-gas chromatographicmass spectrometric analysis
of garlic oil obtained by hydrodistillation. J. Chromatogr. A 1036:
91-93 (2004)
Lee JM, Kim DH, Chang PS, Lee JH. Headspace-solid phase
microextraction (HS-SPME) analysis of oxidized volatiles from free
fatty acids (FFA) and application for measuring hydrogen donating
antioxidant activity. Food Chem. 103: 414-420 (2007)
Bae HJ, Chun HJ. Changes in volatile sulfur compounds of garlic
under various cooking conditions. Korean J. Soc. Food Cookery
Sci. 18: 365-372 (2002)
Imai J, Ide N, Nagae S, Moriguchi T, Matsuura H, Itakura Y.
Antioxidant and radical scavenging effects of aged garlic extract and
its constituents. Planta Med. 60: 417-420 (1994)
Lojzova L, Riddellova K, Hajslova J, Zrostlikova J, Schurek J,
Cajka T. Alternative GC-MS approaches in the analysis of
substituted pyrazines and other volatile aromatic compounds formed
during Maillard reaction in potato chips. Anal. Chim. Acta 641:
101-109 (2009)
Park MH, Jeong MK, Yeu JD, Son HJ, Lim CL, Hong EJ, Noh BS,
Lee JH. Differentiation of sesame oils with diverse roasting
conditions using volatile analysis by a combination of solid phasemicroextraction (SPME) and electronic nose techniques. J. Food
Sci. 76: C80-C88 (2011)
Lee SM, Jo YJ, Kim YS. Investigation of the aroma-active
compounds formed in the Maillard reaction between glutathione and
reducing sugars. J. Agr. Food Chem. 58: 3116-3124 (2010)
Yu TH, Wu CM, Liou YC. Volatile compounds from garlic. J. Agr.
Food Chem. 37: 725-730 (1989)
Zunin P, Salvadeo P, Boggia R, Lanteri S. Study of different kinds
of Pesto Genovese by the analysis of their volatile fraction and
chemometric methods. Food Chem. 114: 306-309 (2009)