Food Chemistry 145 (2014) 126–134

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Evaluation of aroma differences between high-salt liquid-state

fermentation and low-salt solid-state fermentation soy sauces from
China
Yunzi Feng, Yu Cai, Guowan Su, Haifeng Zhao, Chenxia Wang, Mouming Zhao ⇑
College of Light Industry and Food Sciences, South China University of Technology, Guangzhou 510640, China

a r t i c l e i n f o a b s t r a c t

Article history: Two types of Chinese soy sauce, high-salt liquid-state fermentation soy sauce (HLFSS) and low-salt solid-
Received 23 April 2013 state fermentation soy sauce (LSFSS), were used to investigate their differences in aroma profile by head-
Received in revised form 11 July 2013 space solid-phase microextraction (HS-SPME) and gas chromatography-olfactometry/mass spectrometry
Accepted 17 July 2013
(GC-O/MS). Results from descriptive sensory analysis showed that the alcoholic, cooked potato-like and
Available online 31 July 2013
caramel-like attributes were significantly higher in HLFSS, while LSFSS exhibited significantly higher sour
and burnt attributes. In addition, aroma extract dilution analysis (AEDA) revealed 37 and 33 odour-active
Keywords:
regions for HLFSS and LSFSS, respectively. Ethanol, 3-methyl-1-butanol, phenylacetaldehyde, 4-ethyl-2-
Chinese soy sauces
SPME–GC-O
methoxyphenol, 2-methoxy-4-vinylphenol and 3-(methylthio)propanal detected in HLFSS showed the
Key aroma-active compounds highest flavour dilution (FD) factors, while 3-methylbutanal, phenylacetaldehyde and ethyl propanoate
Aroma extract dilution analysis possessed the highest FD factors in LSFSS. Therefore, the traditional Chinese soy sauce HLFSS contained
more complex volatiles and exhibited a richer aromatic profile compared with LSFSS.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction which is considered as the traditional Chinese-type soy sauce.

Soy sauce, which originated in China over 2500 years ago, is the
most widely accepted condiment in Southeast Asian countries,
such as China, Japan, Korea and Thailand (Gao et al., 2010). Its pop-
ularity in the Western part of the world is growing dramatically
due to its unique taste and aroma (Li, Zhao, Zhao, & Cui, 2010).
Nowadays, the annual production of soy sauce in China is approx-
imately 6 million tonnes, accounting for over 65% of the total world
production.
In China, soy sauce can be classified into high-salt liquid-state
fermentation soy sauce (HLFSS) and low-salt solid-state fermenta-
tion soy sauce (LSFSS) according to the different fermentation
process used in its preparation. HLFSS production is carried out
at 15–30 °C by an indigenous fermentation procedure having long
ageing period (90–180 days) and higher brine solution concentra-
tions (17–20%), while LSFSS production employs pure cultures at
relatively high temperature (40–55 °C) having very short aging
(15–30 days) period and lower brine solution concentrations
(13–15%) (Gao et al., 2010; Sun, Jiang, & Zhao, 2012; Zhang &
Tao, 2010). Thus, although LSFSS dominates the market of soy
sauce in China, due to its high production efficiency and low pro-
duction cost, its flavour quality is generally lower than HLFSS,
⇑ Corresponding author. Tel./fax: +86 20 87113914.
E-mail address: femmzhao@scut.edu.cn (M. Zhao).
Moreover, HLFSS has been taking an increasing proportion of the
Chinese market because of its better consumer flavour acceptabil-
ity (Sun et al., 2012). Thus, it is important to get insight into the key
characteristic flavours in both kinds of soy sauce, so as to improve
the flavour quality of soy sauce.
Several studies on the volatile compositions of Chinese soy
sauces have been reported, and results from these studies sug-
gested that aroma profiles of HLFSS and LSFSS are quite different
(Gao et al., 2010; Sun et al., 2012; Zhang & Tao, 2010). Gao et al.
(2010) found that the dominant volatile compound groups of
HLFSS were alcohols, esters, aldehydes and ketones. Sun et al.
(2012) studied the volatiles in twelve samples of HLFSS using
HS-SPME, and found that alcohols and acids were the main vola-
tiles. On the other hand, the predominant volatile groups isolated
in LSFSS were acids and alcohols, accounting for about 47% and
14% of the total area, respectively (Zhang & Tao, 2010). However,
the chemical analysis and quantification of the flavour volatiles
was not satisfactory in these studies.
Gas chromatography-olfactometry (GC-O) has been applied to
identify the aroma-active compounds in many foods (Aceña, Vera,
Guasch, Busto, & Mestres, 2011; Zhao et al., 2011). 4-Hydroxy-5-
ethyl-2-methyl-3(2H)-furanone (4-HEMF), 4-hydroxy-2,5-di-
methyl-3(2H)-furanone (HDF) and 3-(methylthio)propanal were
detected as the most aroma-active compounds with a high flavour
dilution (FD) factor in Japanese and Korean soy sauces (Baek & Kim,

0308-8146/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.foodchem.2013.07.072
 Y. Feng et al. / Food Chemistry 145 (2014) 126–134
2004; Kaneko, Kumazawa, & Nishimura, 2012; Lee, Seo, & Kim,
2006; Steinhaus & Schieberle, 2007; Zhao et al., 2011). Kaneko
et al. (2012) has clarified 25 key aroma compounds in five different
types of Japanese soy sauces, and found that some key aroma-ac-
tive compounds showed significantly different contributions to
these five types of soy sauce. Lee et al. (2006) has reported that
alcohols and pyrazines were the intense aroma-active compounds
in fermented and acid-hydrolysed Korean soy sauce, respectively.
Fifty-six key odorants were detected in all and 16 odorants were
found in both types of soy sauce, whereas 21 and 19 compounds
were detected only in Japanese and Korean soy sauces, respectively
(Baek & Kim, 2004; Kaneko et al., 2012; Lee et al., 2006; Steinhaus
& Schieberle, 2007). The number of aroma-active pyrazines in
Korean soy sauce is higher than those in Japanese soy sauce, while
the esters might be the characteristic key odorants in Japanese soy
sauce.
Different sampling techniques have been developed to collect
aroma prior to GC-O analysis. SPME is a quick, solvent-free, and
quite simple technique that requires very little sample manipula-
tion (Yan, Zhang, Tao, Wang, & Wu, 2008). SPME-AEDA has been
applied to the analysis of aroma compounds on many different
kinds of foods, such as roasted pistachio and soybean paste (Aceña
et al., 2011; Zhao et al., 2011). Furthermore, SPME-AEDA has been
demonstrated to be a good alternative to obtain representative ar-
oma extracts with a wide range of odorants (Aceña, Vera, Guasch,
Busto, & Mestres, 2010).
The aim of this study was to investigate the different sensory
attributes and flavour volatiles in HLFSS and LSFSS, and subse-
quently to identify the aroma-active compounds by aroma extract
dilution anaylsis (AEDA) and GC-O, and then to quantify the most
odour-active compounds from the two types of Chinese soy sauce
and study the effect of these compounds on the whole sensory per-
ception of soy sauce. Results from this study will provide a better
understanding of the key characteristic odorants in HLFSS and
LSFSS to produce soy sauce with high flavour quality.
2. Materials and methods
2.1. Materials
HLFSS samples were purchased from Guangzhou, south China,
while LSFSS samples were obtained from Beijing, north China. All
samples were stored at 4 °C until analysis.
2.2. Chemicals
3-Methylbutanal, 2-methylbutanal, phenylacetaldehyde, 3-
methylbutanoic acid, ethyl isobutyrate, 3-(methylthio)propanal,
ethyl lactate, (E)-2-octenal, phenylglyoxal monohydrate, 4-ethyl-
phenol, 2-acetylpyrrole and isobutyl acetate were purchased from
Sigma–Aldrich (Steinheim, Germany). 1-Propanol, 2,3-butanediol,
3-methyl-1-butanol, 2-methyl-1-butanol, 1-octen-3-ol, 2-ethyl-1-
hexanol, benzyl alcohol, 2-methylpropanal, nonanal, 2,4,7,9-
tetramethyl-5-decyn-4,7-diol, 5-methyl-2-phenyl-2-hexenal, 2-
pentanone, 3-pentanone, 2,3-pentanedione, 3-hydroxy-2-buta-
none, 2,3-butanedione, 5-methyl-2-hexanone, 2-heptanone,
2-methylpropanoic acid, butanoic acid, 2-methylbutanoic acid,
propanoic acid, ethyl acetate, ethyl butanoate, butyl acetate, ethyl
isovalerate, methyl benzoate, ethyl benzoate, diethyl succinate, ethyl
phenylacetate, 2,5-dimethylfuran, furfural, 2-furanmethanol, 5-
methyl-2-furancarboxaldehyde, benzofuran, 4-ethyl-2-methoxyphe-
nol, dimethyl sulfide, dimethyl trisulfide 3-(methylthio)propanol,
2,5-dimethylpyrazine, 2-ethyl-3-methylpyrazine and 2,3,5-trimeth-
ylpyrazine were obtained from Aladdin (Shanghai, China). All other
127
chemicals and solvents were of the highest commercial grade and ob-
tained from Sinopharm Chemical Reagent Co. Ltd. (Shanghai, China).
2.3. Extraction of volatile compounds
The sample preparation and SPME technique were used accord-
ing to the methods of Yan et al. (2008) with some modifications.
The SPME Trisplus automated sampler equipped with a 75 lm
Carboxen/polydimethylsiloxane fibre (CAR/PDMS, Supelco, Inc.,
Bellefonte, PA) was employed for the extraction of volatile com-
pounds in soy sauces. Aliquots (8 mL) of soy sauces were trans-
ferred into 20-mL gas-tight glass vessels (Supelco) and saturated
with 1 g NaCl. Prior to analysis, 20 lL of 2-methyl-3-heptanone
(1.724 mg/L in methanol), as an internal standard, were added
and mixed. After being equilibrated at 45 °C for 20 min, the sample
was extracted with CAR/PDMS fibre for 40 min with continuous
heating and agitation. After extraction, the fibre was inserted into
the GC injector for 3 min to desorb analytes. Each soy sauce sample
was extracted in triplicate. In all cases, the fibres were conditioned
before use by inserting them into the GC injector port for 1 h at
260 °C and then were desorbed for 10 min at 260 °C between injec-
tions to prevent any contamination.
2.4. GC-MS and GC-O analysis
Analysis of the volatiles was performed using Trace GC-MS
system equipped with an Ultra GC, a Trisplus automated sampler
and a quadrupole DSQ II MS (Thermo Finnigan, San Jose, CA).
Separation was performed with a TR-5MS column
(30 m  0.25 mm  0.25 lm, J&W Scientific, Folsom, CA) or a TR-
Wax column (30 m  0.32 mm  0.25 lm, J&W Scientific, Folsom,
CA, USA). The GC-MS conditions in this study were previously re-
ported (Feng et al., 2013). Helium was used as carrier gas with a
flow rate of 1.0 mL/min. The split ratio was 20:1. The analytical
conditions were as follows: the temperature of the column was
maintained at 40 °C for 2 min, ramped to 120 °C at 5 °C/min, hold-
ing for 2 min, and then raised to 220 °C at a rate of 7 °C/min and
held at 220 °C for 5 min. Injection temperature was 250 °C and
the ion source temperature was set at 230 °C. The mass spectrom-
eter was operated in electron impact (EI) mode. The ionisation en-
ergy, detector voltage, scan range and scan rate applied for the
analysis were 70 eV, 350 V, m/z 35–350 and 3.00 scans/s, respec-
tively. Chromatograms and mass spectra were evaluated using
Xcalibur™ software Version 2.0 (Thermo Finnigan, San Jose, CA).
Olfactometry analysis was carried out by three experienced
assessors and repeated twice by each panellist. Sniffing time was
approximately 30 min, and each judge carried out one session
per day. Odour-active compounds were defined as those perceived
at the same retention time (RT) and given similar quality descrip-
tions by at least two of the three panellists. Auxiliary gas for ODO II
(SGE, Melbourne, Australia) was helium at 3.0 on the flow-meter
scale. The sniffing cone was purged with humidified air to help
maintain olfactory sensitivity by reducing dehydration of the mu-
cous membrane in the nasal cavity. Condensation of the effluents
was avoided by heating the transfer-line tubing.
2.5. SPME-AEDA
When working with the SPME, there is no liquid extract because
the analytes are retained on the fibre. Therefore, the usual AEDA
cannot be applied. In this study, the original aroma concentrate
of the soy sauce, extracted by SPME, was stepwise diluted by dif-
ferent split ratios, which varied from 10:1, 50:1, 100:1 to 150:1
(Zhao et al., 2011). This was done by three trained panellists, and
an AEDA was performed three times with respect to each sample.
Timing and description of aromas were recorded by one
128 Y. Feng et al. / Food Chemistry 145 (2014) 126–134
experienced researcher. By definition, the FD factor obtained for
each single odorant in the SPME-AEDA is equal to the highest split
ratio in which the odorant could be perceived at the sniffing-port.
2.6. Identification and quantification analysis
Identification was based on retention indices and mass spectra
of reference standards matching in the standard NIST 08 library.
Positive identifications were achieved by matching linear retention
indices and mass spectra with those of standard reference com-
pounds analysed under the same experimental conditions. When
reference standards were not available, tentative identification
was made according to mass spectra and retention indices match-
ing in the NIST 08 Standard Reference Database and Wiley library.
Retention indices were calculated using a C6–C33 n-alkane series
(Supelco) under the same chromatographic conditions. In some
cases, due to the co-elution of several compounds with similar aro-
matic qualities, their odour threshold constituted a helpful tool to
determine the real contributor.
Semi-quantification measurements were carried out by
2-methyl-3-heptanone equivalents (response factor = 1). For some
volatile compounds (phenylacetaldehyde, ethanol, 3-methyl-1-
butanol, 4-methyl-2-methoxyphenol, 3-(methylthio)propanal,
ethyl propanoate and 3-methylbutanal), chemical aroma standard
mixtures were prepared in aqueous 17% NaCl solution to bracket
the concentrations of each individual compound in soy sauce. Stan-
dard curves according to the internal standard method were cre-
ated for these compounds to obtain more exactly data. Seven
levels of concentration were tested in triplicate. The aroma concen-
trates were analysed by GC-MS in selected-ion-monitoring (SIM)
mode, which provides high sensitivity and precision for the quan-
tification of compounds.
2.7. Sensory evaluation
Quantitative descriptive analysis (QDA) was applied to evaluate
the differences of sensory aroma characteristics among samples
(Gao et al., 2010). Aroma profile characterisation for soy sauce
samples was performed by eight sensory panellists (5 female, 3
male, aged 20–43) recruited from South China University of Tech-
nology, China. The screening procedure was based on the proce-
dure suggested by Steinhaus and Schieberle (2007). The
panellists were subjected to a ranking test with a series of seven
suprathreshold aqueous solutions (25 mL) of ethanol (alcoholic),
acetic acid (sour), 3-methylbutanal (malty) and methional (cooked
potato) and were asked to score the odour intensities. Eight panel-
lists were selected and four 2-h training sessions were held for
descriptor development and definitions in the sensory room at
room temperature (23 ± 2 °C). During the training sessions, the
panellists were exposed to various types of soy sauces. A total of
7 attributes were generated to characterise the sensory properties
of soy sauce samples: ethanol (alcohols), acetic acid (sour), meth-
ional (potato), 4-ethyl-2-methoxyphenol (burnt), phenylacetalde-
hyde (sweet), HEMF (caramel-like), and ethyl acetate (fruity). The
judges scored each attribute on a line scale of 0–9, in which 9
was the highest intensity and 0 was the lowest intensity with no
perception. Fifty-millilitre samples were marked with three-digit
numbers, subsequently put in a plastic cup (100 mL) and covered
with lids, prior to serving for evaluation.
In order to discuss the effects of selected key odorants on fla-
vour descriptors of soy sauce, a further sensory evaluation was
conducted. The soy sauce was spiked with a certain content of indi-
vidual volatiles, and the concentration of added volatile compound
was the same as its original concentration in this type of soy sauce.
In other words, the concentration of the selected key odorant was
doubled in test samples. Then, the panellists were asked to
evaluate the intensities of the odour qualities in each soy sauce
sample and its original control. The positive and negative effects
mean stronger intensities and weaker intensities in the odour attri-
butes, respectively.
2.8. Statistical analysis
Univariate statistical analysis (one-way ANOVA) was applied to
the data to determine significant differences between the soy
sauce samples. A study of the comparison of means was carried
out using Tukey’s test by using the statistical package SPSS 16.0
(SPSS Inc., Chicago, IL).
3. Results and discussion
3.1. Sensory analysis of HLFSS and LSFSS
As shown in Fig. 1, the sensory analysis of the two types of
Chinese soy sauce revealed significant differences for some of the
descriptors evaluated. The aroma profiles differed significantly
(p < 0.05) with regard to the sour, alcohols, cooked potato-like,
caramel-like and burnt descriptors. A higher volatile complexity
in HLFSS was detected by sensory evaluation, which presented a
higher intensity for alcohols, potato-like and caramel-like attri-
butes, while LSFSS showed a higher intensity for sour and burnt.
Furthermore, the perception of sour attributes, especially in LSFSS,
was so high that it exceeded the rest of attributes.
3.2. Comparison of volatile profiles in HLFSS and LSFSS
A total of 92 volatiles were detected in the soy sauce, and 55 of
them were positively identified (Table 1). Some of these com-
pounds have been reported previously (Gao et al., 2010; Kaneko
et al., 2012; Lee et al., 2006; Steinhaus & Schieberle, 2007; Sun
et al., 2012; Yan et al., 2008; Zhang & Tao, 2010). According to
the results of GC-MS, the chromatographic profile obtained in
HLFSS extract was richer than that obtained in LSFSS (data not
shown). As shown in Table 1, the two soy sauces were character-
ised by a different set of volatiles. In detail, alcohols and esters
were the largest groups in HLFSS accounting for approximately
62% and 17% of the total volatiles, respectively. On the other hand,
acids and aldehydes were the dominant classes in LSFSS, and ac-
counted for 43% and 29% of the total volatiles, respectively.
In the alcohols group, ethanol, 2-methyl-1-butanol, 3-methyl-
1-butanol and 2-phenylethanol showed the highest contents in
both soy sauces. The content of ethanol, produced by yeast through
alcoholic fermentation, was 39-fold higher in HLFSS than in LSFSS.
This result might be attributed to the lower fermentation
Fig. 1. Aroma profile analysis of high-salt liquid-state fermentation soy sauce
(black) and low-salt solid-state fermentation soy sauce (grey).
 Y. Feng et al. / Food Chemistry 145 (2014) 126–134 129

Table 1
Volatile compounds identified in Chinese HLFSS and LSFSS.

Compounds RIa Relative peak areab Method of identificationc

TR-5 ms TR-wax HLFSS LSFSS
Alcohols
Ethanol <600 <1000 322 ± 12.6 8.24 ± 0.06 ABC
2-Methyl-1-propanol 629 1091 0.22 ± 0.01 0.03 ± 0.01 ABC
3-Methyl-1-butanol 740 1206 25.3 ± 0.17 4.2 ± 0.18 ABC
2-Methyl-1-butanol 744 1207 16.2 ± 0.97 5.99 ± 0.50 ABC
1-Octen-3-ol 989 1451 1.46 ± 0.01 – ABC
Eucalyptol 1033 – 0.12 ± 0.01 AB
2-Phenylethanol 1131 1893 8.38 ± 0.76 1.72 ± 0.19 ABC
Total 373.91 20.3
Esters
Ethyl acetate 612 <1000 89.6 ± 7.88 4.54 ± 0.41 ABC
Ethyl propanoate 713 <1000 0.47 ± 0.04 1.3 ± 0.12 ABC
Ethyl 2-hydroxypropanoate 824 1339 1.42 ± 0.11 0.18 ± 0.01 ABC
2-Methylbutanoic acid, ethyl ester 850 1049 0.09 ± 0.00 – ABC
3-Methyl-1-butanol, acetate 879 1119 0.16 ± 0.01 0.09 ± 0.01 ABC
Ethyl hexanoate 1003 1231 0.24 ± 0.02 0.11 ± 0.01 ABC
Formic acid phenyl ester 1015 0.22 ± 0.03 0.6 ± 0.07 A
Methyl benzoate 1105 1605 0.47 ± 0.04 0.14 ± 0.01 ABC
Ethyl benzoate 1183 1650 5.47 ± 0.61 1.56 ± 0.18 ABC
Ethyl phenylacetate 1249 1771 0.9 ± 0.10 – ABC
Acetic acid, 2-phenylethyl ester 1263 1799 0.6 ± 0.05 0.28 ± 0.03 ABC
3-Methyl-1-butanol, benzoate 1433 1893 0.51 ± 0.05 0.32 ± 0.02 AB
Propanoic acid, 2-methyl-1-(1.1-dimethylethyl)2-methyl-1.3-propanediyl ester 1605 0.36 ± 0.02 1.01 ± 0.11 AB
Total 101 10.1
Acids
Acetic acid 623 1449 42.7 ± 4.75 33.3 ± 3.39 ABC
Propanoic acid 730 1538 – 0.64 ± 0.05 ABC
2-Methylpropanoic acid 806 1567 – 4.51 ± 0.35 ABC
Butanoic acid 839 1626 – 2.94 ± 0.32 ABC
3-Methylbutanoic acid 882 1666 1.72 ± 0.19 18.41 ± 1.97 ABC
2-Methylbutanoic acid 890 1667 0.81 ± 0.09 9.89 ± 1.01 ABC
Pentanoic acid 926 1735 – 0.63 ± 0.05 AB
3-Methylpentanoic acid 983 1788 – 1.47 ± 0.10 AB
4-Methylpentanoic acid 990 1800 – 3.46 ± 0.38 AB
Sorbic acid 1137 2128 – 34.4 ± 3.27 ABC
Benzoic acid 1209 2418 16.39 ± 1.56 1.55 ± 0.13 ABC
Total 61.6 111
Aldehydes
2-Methylpropanal <600 <1000 2.57 ± 0.22 5.06 ± 0.47 ABC
3-Methylbutanal 655 <1000 6.67 ± 0.45 36.7 ± 3.10 ABC
2-Methylbutanal 665 <1000 19.4 ± 1.73 24.8 ± 2.39 ABC
(E)-2-methyl-2-butenal 748 0.12 ± 0.01 0.46 ± 0.03 AB
Benzaldehyde 980 1506 1.42 ± 0.11 4.37 ± 0.40 ABC
Phenylacetaldehyde 1060 1625 3.32 ± 0.38 2.3 ± 0.27 ABC
Nonanal 1106 1396 – 0.61 ± 0.04 ABC
n-Ethylidene-phenylacetaldehyde 1285 0.11 ± 0.01 0.26 ± 0.03 AB
5-Methyl-2-phenyl-2-hexenal 1498 2053 0.16 ± 0.01 0.14 ± 0.01 ABC
Total 33.8 74.68
Ketones
Acetone <600 <1000 – 4.07 ± 0.45 ABC
2-Butanone 603 <1000 0.16 ± 0.02 0.46 ± 0.05 ABC
3-Methyl-2-pyrrolidinone 709 – 0.22 ± 0.02 A
4-Methyl-2-hexanone 852 0.26 ± 0.02 – ABC
5-Methyl-2-hexanone 862 1136 0.44 ± 0.05 0.13 ± 0.01 ABC
2-Methyl-3-octanone 947 0.17 ± 0.02 0.07 ± 0.01 AB
2,6-Dimethyl-3-heptanone 956 0.42 ± 0.04 0.23 ± 0.02 AB
Total 1.45 5.18
Furan(one)s
2-Methylfuran 603 <1000 0.19 ± 0.02 1.49 ± 0.11 AB
2,5-Dimethylfuran 707 <1000 1.68 ± 0.13 0.17 ± 0.01 ABC
Dihydro-2-methyl-3(2H)-furanone 822 1253 0.21 ± 0.03 1.09 ± 0.07 AB
Furfural 850 1455 1.78 ± 0.10 1.35 ± 0.09 ABC
2-Furanmethanol 873 1654 0.72 ± 0.06 0.38 ± 0.03 ABC
Furfuryl formate 901 2.97 ± 0.35 – AB
2-Acetylfuran 922 0.5 ± 0.03 2.22 ± 0.17 AB
5-Methyl-2-furancarboxaldehyde 980 1562 0.16 ± 0.01 0.74 ± 0.05 ABC
Benzofuran 1006 1487 0.5 ± 0.04 0.14 ± 0.01 ABC
1-(2-furanyl)-1-propanone 1024 0.22 ± 0.01 0.08 ± 0.00 AB
2-Acetyl-5-methylfuran 1052 0.02 ± 0.00 0.19 ± 0.01 AB

(continued on next page)

130 Y. Feng et al. / Food Chemistry 145 (2014) 126–134

Table 1 (continued)

Compounds RIa Relative peak areab Method of identificationc

TR-5 ms TR-wax HLFSS LSFSS
HEMFd 1180 2080 0.05 ± 0.01 0.01 ± 0.00 ABC
3-Phenylfuran 1235 1833 1.38 ± 0.16 0.26 ± 0.02 AB
Total 10.38 8.12
Sulfur-containing compounds
Dimethyl sulfide <600 <1000 0.05 ± 0.01 – ABC
Dimethyl disulfide 744 1062 0.79 ± 0.07 0.56 ± 0.06 ABC
2-Methylthiophene 774 1082 – 1.29 ± 0.11 AB
S-methyl 3-methylbutanethioate 919 – 0.22 ± 0.02 AB
3-(Methylthio)propanal (methional) 923 1443 0.65 ± 0.05 0.05 ± 0.01 ABC
Dimethyl trisulfide 978 1357 0.21 ± 0.01 0.07 ± 0.00 ABC
5-Methyl-2-furanmethanethiol 1001 1359 0.66 ± 0.08 – AB
4-(1,1-Dimethylethyl)-2-methylbenzenethiol 1452 0.25 ± 0.02 0.37 ± 0.03 AB
Total 2.62 2.55
Phenols
2-Methoxyphenol 1099 1843 1.89 ± 0.05 0.97 ± 0.03 ABC
4-Ethylphenol 1196 2158 0.2 ± 0.02 0.25 ± 0.01 ABC
4-Ethyl-2-methoxyphenol 1286 2010 0.29 ± 0.02 0.02 ± 0.01 ABC
2-Methoxy-4-vinylphenol 1330 2170 0.22 ± 0.01 – ABC
Total 2.58 1.24
Pyrazines
2-Methylpyrazine 838 1258 0.1 ± 0.01 1.31 ± 0.12 ABC
2,6-Dimethylpyrazine 922 1320 0.66 ± 0.06 1.23 ± 0.12 ABC
2-Ethyl-6-methylpyrazine 1010 1375 0.5 ± 0.05 0.19 ± 0.01 ABC
Trimethylpyrazine 1015 1396 0.39 ± 0.04 0.37 ± 0.02 ABC
3-Ethyl-2,5-dimethylpyrazine 1084 0.16 ± 0.01 0.08 ± 0.01 AB
2-Isobutyl-3-methylpyrazine 1244 0.31 ± 0.02 – AB
2-Isoamyl-6-methylpyrazine 1253 1591 0.32 ± 0.03 – AB
Total 2.44 3.18
Others
Toluene 767 1032 0.17 ± 0.01 0.11 ± 0.01 AB
Styrene 895 0.1 ± 0.01 0.07 ± 0.01 AB
Unknown 906 0.13 ± 0.01 0.25 ± 0.02 AB
Unknown 974 0.8 ± 0.09 3.72 ± 0.41 AB
Limonene 1028 1186 0.21 ± 0.02 0.16 ± 0.02 AB
3-Acetyl-1H-pyrroline 1089 1951 3.27 ± 0.31 8.55 ± 0.81 ABC
3-Hydroxy-2-methyl-4-pyranone (maltol) 1142 1950 0.05 ± 0.01 0.10 ± 0.01 ABC
Naphthalene 1201 1711 2.93 ± 0.25 3.01 ± 0.28 AB
Unknown 1244 – 0.21 ± 0.02 AB
1,2-Dihydro-1,5,8-trimethyl-naphthalene 1353 0.28 ± 0.02 – AB
Di-epi-cedrene 1396 1541 – 1.69 ± 0.18 AB
Cedr-8(15)-ene 1404 0.63 ± 0.05 1.06 ± 0.1 AB
Cedrol 1632 2085 1.09 ± 0.1 1.62 ± 0.14 AB
Total 9.54 20.54
a
Retention indices calculated on TR-5 ms and TR-wax capillary column, respectively.
b
Relative peak areas to that of internal standard (20 ll of 1.724 mg/L 2-methyl-3-heptanone in methanol) using TR-5 ms column. Values expressed as average
(n = 3) ± standard deviation; –, not detected.
c
Method of identification: (A) by comparison of the mass spectrum with the NIST/Wiley mass spectral library; (B) by comparison of RI (Kovats indices) with RI of an
authentic compound; and (C), by comparison of retention time and spectrum of an authentic compound.
d
HEMF, 4-hydroxy-5-ethyl-2-methyl-3(2H)-furanone.

temperature and more active alcoholic fermentation of yeast cells
in the moromi fermentation stage of HLFSS. Indeed, Wu et al.
(2012) have verified that the soy sauce would have lower concen-
tration of ethanol when higher fermentation temperature was
used. This could be explained by the evaporation of ethanol during
the fermentation, especially under the high temperature in the
LSFSS process (Abdel-Banat, Hoshida, Ano, Nonklang, & Akada,
2009). Furthermore, ethanol has been detected previously only in
one of the five LSFSS samples and the content was relatively low
(Zhang & Tao, 2010), which was in agreement with the results from
this study. Thus, ethanol might be a discriminating compound for
HLFSS. Moreover, fusel alcohols (2-methyl-1-butanol, 3-methyl-
1-butanol and 2-phenylethanol) were identified in appreciable
abundance in HLFSS in this study. These compounds, from
branched-chain amino acids isoleucine and leucine and aromatic
amino acid phenylalanine, respectively, were mainly produced
during the long moromi fermentation stage (Feng et al., 2013).
Esters are one of the most important aroma compounds in fer-
mented foods due to their high volatility and sensitivity to the
olfactory receptors of human (Lee & Ahn, 2009; Steinhaus &
Schieberle, 2007). The concentrations of esters, especially ethyl ace-
tate, were obviously higher in HLFSS than in LSFSS. The results could
be explained by the shorter maturation time in LSFSS, which was not
suitable for accumulation of ethyl esters (Zhao et al., 2011).
A distinct separation of volatile acid class was observed be-
tween HLFSS and LSFSS. Among the 11 volatiles in the group of
acids, only 4 acids (acetic acid, 3-methylbutanoic acid, 2-methylb-
utanoic acid and benzoic acid) were found in both soy sauces. Se-
ven other volatile acids were only detected in LSFSS including
propanoic acid, 2-methylpropanoic acid, butanoic acid, pentanoic
acid, 3-methylpentanoic acid, 4-methylpentanoic acid and sorbic
acid. Moreover, the percentage of volatile acids group was approx-
imately half of the total volatiles in LSFSS. These acid compounds
might be the major contributors to the strong sour note in LSFSS.
 Y. Feng et al. / Food Chemistry 145 (2014) 126–134 131

Table 2
Aroma-active compounds of two kinds of soy sauce.

LRIa Odor qualityb Compounds HLFSS LSFSS

1060 Honey-like Phenylacetaldehyde 1:150 1:150
<600 Alcoholic, solvent-like Ethanol 1:150 1:10
740 Malty 3-Methyl-1-butanol 1:150 1:10
1286 Smoky, bacon, soy sauce 4-Ethyl-2-methoxyphenol 1:150 1:10
923 Cooked potato 3-(Methylthio)propanal (methional) 1:150 ndd
1330 Spicy, burnt 2-Methoxy-4-vinylphenolc 1:150 nd
713 Fruity Ethyl propanoate 1:100 1:150
665 Malty, almond 2-Methylbutanal 1:100 1:100
850 Bread, almond, sweet Furfural 1:100 1:100
1209 Fruity, floral Benzoic acid 1:100 1:100
612 Fruity Ethyl acetate 1:100 1:50
1180 Caramel-like 4-Hydroxy-5-ethyl-2-methyl-3(2H)-furanone 1:100 1:50
824 Sweet, fatty Ethyl 2-hydroxypropanoate 1:100 1:10
873 Fermented sugar 2-Furanmethanol 1:100 1:10
1105 Flowery, honey Methyl benzoate 1:100 1:10
1131 floral, sweet 2-phenylethanol 1:100 1:10
707 Solvent-like 2,5-Dimethylfuran 1:100 nd
906 Roasty Unknown 1:100 nd
1034 Fresh hazelnut-like 2-Ethenyl-6-methyl-pyrazinec 1:100 nd
655 Malty, almond 3-Methylbutanal 1:50 1:150
<600 Malty, nutty 2-Methylpropanal 1:50 1:50
1089 Roasty, popcom-like 3-Acetyl-1H-pyrroline 1:50 1:50
1396 Sweet Di-epi-cedrenec 1:50 1:10
1253 Rubbery, sweet 2-Isoamyl-6-methylpyrazinec 1:50 nd
838 Nutty, popcorn 2-Methylpyrazinec 1:10 1:100
806 Cheese, fatty 2-Methylpropanoic acid 1:10 1:50
974 Sweet, soy sauce-like Unknown 1:10 1:50
1052 Nutty aroma, cocoa-like 2-Acetyl-5-methylfuranc 1:10 1:50
822 Caramel Dihydro-2-methyl-3(2H)-furanonec 1:10 1:50
623 Sour Acetic acid 1:10 1:10
1015 Burnt 2,3,5-Trimethylpyrazine 1:10 1:10
1142 Caramel, sweet 3-Hydroxy-2-methyl-4-pyranone (maltol) 1:10 1:10
1235 Green bean-like 3-Phenylfuranc 1:10 1:10
744 Onion, cooked cabbage Dimethyl disulfidec 1:10 nd
978 Sulfury, cooked onion Dimethyl trisulfide 1:10 nd
1004 Cooked potato 3-(Methylthio)-1-propanol 1:10 nd
1263 Honey, rosy Acetic acid, 2-phenylethyl ester 1:10 nd
774 Sulfur, cooked vegetables 2-Methylthiophenec nd 1:100
919 Cheese, soup S-methyl 3-methylbutanethioatec nd 1:50
983 Sour herb, slightly green grass 3-Methylpentanoic acidc nd 1:50
882 Cheese-like, sweaty 3-Methylbutanoic acid nd 1:10
890 Cheese-like, sweaty 2-Methylbutanoic acid nd 1:10
980 Almond, spicy, caramel 5-Methyl-2-furancarboxaldehyde nd 1:10
a
Retention index on TR-5 ms column.
b
Odour descriptions are based on (I) the University of Florida Citrus Flavor database (http://www.crec.ifas.ufl.edu/rouseff/#) and (II) Flavornet (http://www.flavornet.org/
flavornet.html).
c
Tentatively identified compounds.
d
nd, not detected.

This result was in agreement with the sensory evaluation (Fig. 1),
where the perception of sour attribute was so high that it exceeded
the rest of attributes in LSFSS.
Aldehydes represented 6% and 29% of the total volatile compo-
sition in HLFSS and LSFSS, respectively. The major aldehydes (3-
methylbutanal, 2-methylbutanal, 2-methylpropanal, benzaldehyde
and phenylacetaldehyde), contributing to the desirable aroma of
soy sauce, have also been described in many soybean fermented
foods, such as Korean soybean pastes and Thai thua nao (Dajanta,
Apichartsrangkoon, & Chukeatirote, 2011; Lee & Ahn, 2009; Sun
et al., 2012).
With regards to the minor volatile classes in Table 1, phenols,
furanones, sulfur-containing compounds and pyrazines play
important roles in the characteristic aroma of soy sauce (Steinhaus
& Schieberle, 2007). The concentration of phenols in HLFSS, includ-
ing 2-methoxyphenol, 4-ethylphenol, 4-ethyl-2-methoxyphenol
and 2-methoxy-4-vinylphenol, was obviously higher than in LSFSS.
On the other hand, similar contents of sulfur-containing com-
pounds were found in the two types of Chinese soy sauces. How-
ever, HLFSS exhibited different sulfur-containing compounds
compared with LSFSS; dimethyl sulfide and 5-methyl-2-furan-
methanethiol were only detected in HLFSS, while 2-methylthioph-
ene and S-methyl-3-methylbutanethioate were only found in
LSFSS.
3.3. Identification of aroma-active compounds in HLFSS and LSFSS
A total of 43 aroma-active compounds were found by using
SPME-GC-O coupled with GC-MS from the two types of Chinese
soy sauce (Table 2). It included 5 esters, 6 acids, 4 aldehydes, 3
alcohols, 4 pyrazines, 8 furanones, 6 sulfur-containing compounds,
2 phenols, 1 pyranone, 1 pyrrole, 1 hydrocarbon and 2 unidentified
compounds. Among these compounds, 10 aroma-active regions
were detected with the same FD factor for HLFSS and LSFSS. There
were phenylacetaldehyde (FD 1:150), furfural, 2-methylbutanal,
benzoic acid (FD 1:100), 2-methylpropanal, 2-acetyl-1H-pyrrole
(FD 1:50), acetic acid, 2,3,5-trimethylpyrazine, maltol and 3-phen-
ylfuran (FD 1:10). When analysing the differences between varie-
ties, 10 aroma-active regions including 2-methoxy-4-vinylphenol,
2,5-dimethylfuran, 3-(methylthio)propanal, 2-ethenyl-6-methyl-
pyrazine, 2-isoamyl-6-methylpyrazine, dimethyl disulfide, di-
methyl trisulfide, 3-(methylthio)-1-propanol, phenethyl acetate
132 Y. Feng et al. / Food Chemistry 145 (2014) 126–134
Table 3
Concentrations, thresholds, and odour activity values (OAV) of the most intense odorants (split ratio 1:150) identified in Chinese soy sauces.
Name m/za Concentrations (lg/L)
HLFSS
Phenylacetaldehyde 91 230
Ethanol 46 11462667
3-Methyl-1-butanol 55 2937
4-Ethyl-2-methoxyphenol 137 47
3-(Methylthio)propanal 48 412
2-Methoxy-4-vinylphenolc 150 36
Ethyl propanoate 57 15
3-Methylbutanal 44 950
a
Selected ions (m/z) used in quantitative analysis, which was carried out with calibration curves based on standards.
b
Calculated by dividing the concentration by the odour threshold in water.
c
Due to the absence of pure standards, 2-methoxy-4-vinylphenol was quantified by the standard curves of similar structure compound 4-ethyl-2-methoxyphenol.
d
Odour threshold taken from Steinhaus and Schieberle (2007).
e
Odour threshold taken from Giri, Osako, Okamoto, and Ohshima (2010).
and an unknown compound were detected for HLFSS but not for
LSFSS. These compounds could contribute to the aroma of ‘‘cooked
potato’’, ‘‘onion’’, ‘‘nutty’’ or ‘‘roasty’’ in HLFSS. On the contrary, 2-
methylthiophene (sulfur, cooked vegetables), S-methyl-3-meth-
ylbutanethioate (cheese, soup), 3-methylpentanoic acid (sour
herb), 3-methylbutanoic acid (cheese-like), 2-methylbutanoic acid
(cheese-like) and 5-methyl-2-furancarboxaldehyde (almond,
spicy) were described as aromatic components only in LSFSS. It
should be noted that acids class exhibited elevated concentrations
in LSFSS.
Early GC-O studies on Korean and Japanese soy sauces have re-
ported 11–24 and 25–30 aroma-active volatiles, respectively (Baek
& Kim, 2004; Kaneko et al., 2012; Lee et al., 2006; Steinhaus &
Schieberle, 2007). Most of the soy sauce aroma-active volatiles ob-
served in this study have been previously reported (Baek & Kim,
2004; Kaneko et al., 2012; Lee et al., 2006; Steinhaus & Schieberle,
2007). However, the overall aroma profiles of Chinese soy sauce
were significantly different from those of Korean and Japanese
soy sauces. Aroma-active volatiles detected in earlier studies (Baek
& Kim, 2004; Kaneko et al., 2012; Lee et al., 2006; Steinhaus &
Schieberle, 2007) such as 4-hydroxy-2,5-dimethyl-3(2H)-furanone
(HDF), 2,6-dimethoxyphenol, (E)-4,5-epoxy-(E)-2-decenal and (E)-
<beta>-damascenone were not observed in this study. On the other
hand, 3-methylpentanoic acid, di-epi-cedrene, S-methyl 3-meth-
ylbutanethioate, 2-methylthiophene and 5-methyl-2-furancarbox-
aldehyde were only found in Chinese soy sauces.
The volatiles in Table 2 are ranked in order of decreasing aroma
intensity first based on HLFSS soy sauce and then on LSFSS. To eval-
uate the key aroma compounds in Chinese soy sauces, the AEDA by
using different split ratio (1:150, 1:100, 1:50 and 1:10) was ap-
plied. Furthermore, the odour activity value (OAV) concept was
used for the most intense aroma-active volatile compounds in this
study to verify the aroma potency of these individual soy sauce
odorants effectively (Table 3).
3.3.1. HLFSS
As summarised in Table 2, the AEDA yielded 37 odour regions,
which appeared to contribute to the overall aroma, and nearly half
of them showed high FD values (6 compounds for split ratio 1:150
and 13 for split ratio 1:100). Phenylacetaldehyde, ethanol,
3-methyl-1-butanol, 4-ethyl-2-methoxyphenol, 2-methoxy-4-
vinylphenol and 3-(methylthio)propanal could be considered as
important contributors to the aroma of HLFSS because they pos-
sessed the highest FD values.
Phenylacetaldehyde, with a sweet and honey-like note, has
been found in awide variety of fermented soybean foods, such as
sufu and soybean pastes (Chung, Fung, & Kim, 2005; Lee & Ahn,
2009). It is considered as the character impact compound of
Thresholds (lg/L) OAVb
LSFSS HLFSS LSFSS
165 4d 57 41
81666 100000d 115 1
708 4e 734 177
4 50e 1 <1
35 1.4d 294 25
1 3e 12 <1
52 10e 2 5
4602 1.2d 792 3835
Japanese soy sauce (Steinhaus & Schieberle, 2007). In contrast, eth-
anol has been seldom detected as an aroma-active compound in
previous studies. Because the most popular preparation method
used for AEDA is liquid-liquid extraction (LLE) using diethyl ether
as the solvent, the odour of ethanol might be missed when avoid-
ing the harmful effect of solvent peak on the panellists’ health dur-
ing the sniffing test. However, ethanol played an important role in
soy sauce aroma. It was considered as one of 13 key aroma-active
compounds in a model aroma mixture of soy sauce and showed a
high odour activity value (Steinhaus & Schieberle, 2007).
Methional (split ratio 1:150) and methionol (split ratio 1:10),
exhibiting the cooked potato note, are well-known to be generated
from methionine catabolism. The differences detected in the
cooked potato-like note in sensory evaluation were well in line
with the FD factors of methional and methionol, which were only
perceived in HLFSS.
Among the 13 aroma-active regions with a FD factor of 1:100
in HLFSS, most compounds exhibited sweet and fruity odour, such
as 2-phenylethanol, 2-furanmethhanol, ethyl acetate, ethyl
2-hydroxypropanoate, methyl benzoate and ethyl propanoate.
Consequently, the sweet and fruity notes, which are significantly
higher in HLFSS (Fig. 1), might be caused by these compounds.
Besides the abovementioned alcohols and esters, 4-hydroxy-5-
ethyl-2-methyl-3(2H)-furanone (HEMF) should also be paid more
attention. It has been identified as the most intense compound
with highest FD value in previous studies (Kaneko et al., 2012).
However, HEMF was detected with medium aroma intensity in
the present study. This result could be explained by different raw
materials and manufacturing processes employed. In addition,
we found that the peak area of HEMF was relatively low in the
chromatograms indicating weak affinity of HEMF to SPME fibres.
This might be another reason for the different FD values in Chinese
and Japanese soy sauces. Therefore, further research is necessary to
enable a detailed discussion of this point. In this study, the amount
of HEMF in HLFSS was higher than that in LSFSS, which contributed
to a higher caramel-like attribute in HLFSS. In addition, Zhang and
Tao, (2010) did not detect any HEMF in 5 LSFSS samples. The HEMF
in soy sauce could be formed by a combination of a 5-carbon com-
pound generated by the amino-carbonyl reaction under mild con-
ditions and a 2 carbon compound from glucose metabolism by
yeast (Ohata, Kohama, Morimitsu, Kubota, & Sugawara, 2007).
Thus, the difference of HEMF contents between two types of soy
sauce was likely to be associated with a higher level of yeast con-
centration during moromi fermentation of HLFSS.
3.3.2. LSFSS
Regarding LSFSS, 33 aroma-active regions were perceived
(Table 2). In the case of the pronounced sour note, more aroma-
 Y. Feng et al. / Food Chemistry 145 (2014) 126–134
Table 4
Effects of key aroma-active compounds on the soy sauce sensory attributes.
Positive effect
Added in HLFSSa
Phenylacetaldehyde Floral**, sweet **,c
Ethanol Alcoholic***, caramel-like**
3-Methyl-1-butanol Sour**, caramel-like**
4-Ethyl-2-methoxyphenol Sour*, burnt**, full-
flavoured*
3-(Methylthio)propanal Cooked potato**, caramel-
(methional) like**
Added in LSFSSb
Phenylacetaldehyde Floral**
3-Methylbutanal Caramel-like**, malty**, off
odour*
Ethyl propanoate Alcoholic*, fruity**
a
HLFSS, high-salt liquid-state fermentation soy sauce.
b
LSFSS, low-salt solid-state fermentation soy sauce.
c
sensory attributes, having significantly differences between the sample and the
control, were listed in this table.
*
p < 0.1.
**
p < 0.05.
***
p < 0.01).
active acids or compounds with sour aroma in LSFSS were expected
(Fig. 1). Indeed, the area percentage of volatile acids was approxi-
mately 50%. The significantly higher sour note in LSFSS might be
caused by different FD factors of 2-methylbutanoic acid (cheese-
like, sweaty), 3-methylbutanoic acid (cheese-like, sweaty), 2-
methylpropanoic acid (cheese, fatty) and 3-methylpentanoic acid
(sour herb). Except for 2-methylpropanoic acid, none of these acids
were detected in HLFSS using GC-O. It should be noted that 3-
methylpentanoic acid was identified in all LSFSS samples (Zhang
& Tao, 2010). Therefore, 3-methylpentanoic acid could be consid-
ered as a characteristic marker for LSFSS.
Overall, 8 odorants, including 3-methylbutanal, ethyl propano-
ate, furfural, phenylacetaldehyde, 2-methylbutanal, benzoic acid,
2-methylpyrazine, 2-methylthiophene, with FD factors above split
ratio 1:100, were considered as aroma-active volatiles in soy sauce.
Among these compounds, significant differences were found in the
aroma intensity of 2-methylthiophene (sulfur, cooked vegetables),
2-methylpyrazine (nutty, popcorn) and 3-methylbutanal (malty),
which were stronger in LSFSS. To the best of our knowledge, 2-
methylthiophene has not been reported as an aroma-active com-
pound in soy sauce, but has been detected in Chardonnay dry
white wine and extruded potato snacks (Li, Tao, Wang, & Zhang,
2008; Majcher & Jelen, 2007). This compound seemed to be a char-
acteristic compound in LSFSS not occurring in other varieties of soy
sauces. On the other hand, the highest OAV of 3835 was calculated
for 3-methylbutanal (Table 3), a malty-smelling compound pro-
duced by Strecker degradation (Steinhaus & Schieberle, 2007).
Higher FD factors in LSFSS were observed for the Strecker alde-
hydes 2-methylbutanal and phenylacetaldehyde, as well as 3-
methylbutanal, suggesting that the degradation of free amino acids
via the Strecker reaction played an important role in the aroma
compound generation.
3.4. The effects of selected key odorants on the sensory profiles of soy
sauce
To better understand the effect of selected key odorants on the
perception of soy sauce aroma, a preliminary sensory evaluation
test was conducted. The effects of spiking individual volatiles alone
on flavour descriptors are shown in Table 4. Five and three aroma-
active compounds with the highest FD factors (split ratio 1:150) in
HLFSS and LSFSS, respectively, were evaluated in this study.
133
As shown in Table 4, all the treatments were significant differ-
ent from the unspiked control, which further verified the impor-
Negative tance of these key odorants in soy sauce. The effect of real food
effect matrix on sensory perception of the individual key odorant was
obviously distinguished, and more subtle interactions between
Sour* sensory attributes have been observed in the present study. For
example, adding phenylacetaldehyde alone to HLSFF not only im-
parted an intense floral aroma, but also increased sweet aroma
and suppressed sour note. Furthermore, caramel-like attribute
was easily influenced by the added volatiles, even those com-
pounds that did not possess caramel-like odour in water, such as
ethanol and 3-methyl-1-butanol. Additionally, it is worth noting
Sour*** that addition of any selected key odorants would have negative ef-
Sour**
fect on sour note in LSFSS. This might be due to the prominent con-
Sour* tribution of sour attribute to LSFSS (Fig. 1).
4. Conclusions
The present study has clearly demonstrated the significant dif-
ferences in aroma profile and sensory perception between HLFSS
and LSFSS. Thirty-seven and 33 different odorants have been found
in the GC-O profiles for HLFSS and LSFSS, respectively. HLFSS
exhibited a more complex volatile composition than LSFSS. Accord-
ing to the sensory evaluation, HLFSS presented a higher intensity
for alcohols, cooked potato-like and caramel-like notes, while
LSFSS showed a higher intensity for sour and burnt notes. Results
from AEDA experiments showed that phenylacetaldehyde, ethanol,
3-methyl-1-butanol, 4-ethyl-2-methoxyphenol, 3-(methyl-
thio)propanal and 2-methoxy4-vinylphenol were the most intense
aroma-active compounds in HLFSS, while phenylacetaldehyde, 3-
methylbutanal, and ethyl propanoate were the key odorants in
LSFSS. This research is part of our continuous efforts to improve
soy sauce flavour quality by optimising the manufacturing process.
On the basis of results obtained from this study, further work on
conducting aroma recombination experiments and omission tests
are in progress to verify the obtained results and to characterise
the fingerprint of volatile aroma constituents of Chinese soy
sauces.
Acknowledgements
The authors gratefully acknowledge the National Science Tech-
nology Supporting Project for 12th Five-Year Plan (Nos.
2012BAD34B03 and 2012BAK17B11), the National High Technol-
ogy Research and Development Program of China (863 Program)
(Nos. SS2012AA022502 and SS2013AA102106) and the Key
Technology R&D Program of Guangdong Province (Nos.
2011A020102001 and 2012cxy004) for their financial supports.
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