JFS:
Food Chemistry and Toxicology
Volatile Substances of Chinese Traditional
Jinhua Ham and Cantonese Sausage
Abstract: The relationship between volatile substances and manufacturing procedures of Chinese traditional Jinhua
ham and Cantonese sausage were determined. Ethanol and its alcohol esters were the major volatiles of Cantonese
sausage suggesting that ethanol added to meat at the formulation stage was the most important factor for the
characteristic flavor of Cantonese sausage. Small amounts of fatty acid and amino acid breakdown products were
also detected in Cantonese sausage. In Jinhua ham, alkanes and alkenes were the major volatiles, but branched
aldehydes and sulfur compounds were also detected. This indicated that many complicated chemical reactions
occurring in Jinhua ham during ripening contribute to its characteristic flavor.
Keywords: volatile composition, meat, jinhua ham, Cantonese sausage
Introduction
T
HERE ARE NUMEROUS TRADITIONAL
Chinese-style semi-dry sausages in
Chinese traditional market and found
that they varied in thiobarbituric acid
reactive substances ( TBARS) values
and nitrite concentrations. There is no
similar product of Chinese-style semidry sausages in other parts of the
world. The large quantity of wine added into Cantonese sausage during
manufacturing may contribute much
of its specific flavor to the product.
No lactic acid bacteria starter culture
is added to Cantonese sausage, which
therefore differs from most fermented sausages elsewhere. Addition of
large amounts of sugar may also contribute to the flavor formation in the
Chinese-style sausage products. Lin
and others (2000) showed that adding
7 to 9% of sugar in Chinese-style sausage was conducive to proteolysis
than adding 5% of sugar. So far, almost no published report on the volatiles of Cantonese sausage is available.
The aim of this study was to determine the volatile substances of these 2
important traditional Chinese meat
products using purge-and-trap dynamic headspace/gas chromatography-mass spectrometry (GC-MS) and
to assess the possible relationship between these volatiles and manufacturing procedures of these meat products. Further, this work may help
stimulate interest in studies on traditional Chinese-style meat products.
meat products in China and Southeast Asia, but Jinhua ham and Cantonese sausage are the most famous
and popular (Dong and others 1999).
These products have strong characteristic flavors enjoyed by most Chinese
consumers. The characteristic flavors
of these products are considered to be
derived from their volatile substances.
However, almost no information on
the characteristics of these substances
is available.
Jinhua ham is a dry-cured ham
produced through a long ripening period during which lipids and proteins
are degraded by natural enzymes in
meat, producing free fatty acids, free
amino acids and volatile compounds
( Wagu and others 1995; Berdague and
others 1993a). Jinhua ham is quite
similar to Western-style dry-cured
hams where numerous reports have
been published on the volatiles of
those products (Berdague and others
1991; Garcia and others 1991; Barbieri
and others 1992; Sabio and others
1998). Differences in volatile substances between Jinhua and Westernstyle dry-cured hams, however, are
expected because of different manufacturing procedures.
Chinese Cantonese sausage is one
of the most famous Chinese-style
semi-dry sausages. The common ingredients in Chinese-style semi-dry
Materials the Methods
sausages are wine and sugar, added in
large quantities. The amounts of wine
and sugar added, however, vary de- Sample preparation
pending upon the fermentation conFour packages each of Jinhua ham
ditions and the addition of spices. (Zhejiang Cereals, Oil & Foodstuffs I/E
Chou and Liu (1996) surveyed the Corp. Ltd., Hangzhou, China) and
© 2001 Institute of Food Technologists
Cantonese sausage (Guangdong Foodstuffs I/E Corp., Guangdong, China)
were purchased from different stores
in Beijing, China. The bags of ham and
sausage were cut open, divided into
small portions, repackaged in oxygenimpermeable vacuum bags (O 2 permeability, 9.3 mL O2/m2/24 h at 0 8C), and
then stored at –20 8C until used. Before
analysis, samples were stored at 4 8C
overnight.
Lipid extraction
Lipids were extracted from samples
according to the method of Folch and
others (1957). Two grams of ham or sausage sample, butylated hydroxytoluene
(BHT; 50 mL, 7.2%), and 30 mL Folch 1
solution (chloroform:methanol = 2:1)
were added to a 50-mL test tube and homogenized with 3 volumes of deionized
distilled water using a Polytron (Brinkman Instruments, Inc., Westbury, N.Y.,
U.S.A.) for 20 s at high speed. The homogenate was filtered through a Whatman
No. 1 filter paper (Whatman Inc., Clifton, N.J., U.S.A.) into a 100-mL graduated
cylinder, and the filter paper was rinsed
twice with 10 mL of Folch 1 solution. After addition of 8 mL of 0.88% NaCl solution to each cylinder, the cylinder was
capped with a glass stopper and the content mixed. The inside of the cylinder
was washed twice with 2 mL of Folch 2
solution (chloroform: methanol: water = 3:47:48). After phase separation, the
lipid layer volume was recorded, and the
upper layer (methanol and water) of the
solution was completely and carefully siphoned off so as not to contaminate the
chloroform layer. This organic layer was
put in a glass scintillation vial and dried
under nitrogen flow.
Vol. 66, No. 6, 2001—JOURNAL OF FOOD SCIENCE
827
Food Chemistry and Toxicology
M. DU AND D.U. AHN
Volatiles of Chinese Ham and Sausage . . .
Food Chemistry and Toxicology
GC determination of fatty acid
composition
Table 1—Fatty acid composition and TBARS values of Chinese Cantonese sausage and Jinhua1 ham
Twenty-five mg of extracted oil,
transferred into a 20-mL test tube with
cap, was used for methylation. One mL
of methylating reagent (boron trifluoride-methanol; Sigma Chemical Co. St.
Louis, Mo., U.S.A.) was added and incubated in a 90 8C water bath for 1 h. After
this was cooled to room temperature, 2
mL hexane and 5 mL water were added,
mixed thoroughly, and left at room
temperature overnight for phase separation. The top hexane layer containing
methylated fatty acids was used for fatty acid analysis using a GC (HP 6890;
Hewlett Packard Co. Wilmington, Del.,
U.S.A.). A ramped oven temperature
condition (180 8C for 2.5 min, increased
to 230 8C at 2.5 8C/min, then held at
230 8C for 7.5 min) was used. Temperatures at both the inlet and detector
were 280 8C. Helium was the carrier gas
at linear flow of 1.1 mL/min. Flame ionization detector air, H 2, and make-up
gas (He) flows were 350, 35, and 43 mL/
min, respectively. Fatty acids were identified by MS and GC retention times
were compared with known standards.
Relative quantities were expressed as
wt% of total fatty acids.
Item
Volatile analysis
A purge-and-trap dynamic headspace GC/MS system was used to identify and quantify the volatile compounds.
One-half gram of ham or sausage sample was placed in a 40-mL sample vial,
and the vial was flushed with helium gas
(99.999%) for 5 s at 40 psi. After capping
with a Teflon-lined, open-mouth cap,
the vial was placed in a refrigerated
(4 8C) sample tray. The maximum sample holding time in the sample tray before determination of volatiles was less
than 10 h to minimize oxidative changes
(Ahn and others 1999).
The meat sample was purged with
helium gas (40 mL/min) for 15 min.
Volatiles were trapped at 20 8C using a
Tenax/silica gel/charcoal column (Tekmar-Dohrman, Cincinnati, Ohio,
U.S.A.) and desorbed for 2 min at
220 8C. The desorbed volatiles were
concentrated at –90 8C using a cryofocusing unit (Tekmar-Dohrman, Cincinnati, OH), thermally desorbed and injected (30 s) onto a capillary column
by increasing the temperature to
220 8C. An HP-624 column (7.5 m, 0.25
mm i.d., 1.4 mm nominal) and an HP-1
column (52.5 m, 0.25 mm i.d., 0.25 mm
nominal; Hewlett-Packard Co.) were
connected using a zero dead-volume
Jinhua ham
Cantonese sausage
Identification
Total fat content
20.84 6 0.14
31.73 6 0.20
Fatty acid composition
———————— (%) ————————
Tetradecanoic
1.22 6 0.08
1.21 6 0.08
Hexadecenoic
2.34 6 0.31
2.51 6 0.07
Hexadecanoic
23.25 6 0.75
22.86 6 0.22
Heptadecenoic
0.22 6 0.04
0.22 6 0.04
Heptadecanoic
0.24 6 0.06
0.23 ± 0.06
Octadecadienoic
8.98 6 1.40
9.12 ± 1.54
Octadecenoic
44.07 6 2.19
44.55 6 1.44
Octadecatrienoic
3.90 6 0.54
4.23 6 0.23
Octadecanoic
12.39 6 1.29
11.66 6 0.26
Eicosatetrenoic
1.36 6 0.25
1.35 6 0.20
Unidentified
2.03 6 0.28
2.06 6 0.27
Total saturated fatty acid
37.10
35.96
Total unsaturated fatty acid
60.87
61.98
TBARS value
———mg malonaldyde/kg meat———
1.14 6 0.12
4.78 6 0.32
MS/RT2
MS/RT
MS/RT
MS
MS/RT
MS/RT
MS/RT
MS/RT
MS/RT
MS/RT
1n=4. 2 MS: identified by mass spectrometry; RT: identified by retention time of standards.
column connector ( J&W Scientific,
Folsom, Calif., U.S.A.). Ramped oven
temperature was used to improve volatile separation. The initial oven temperature of 0 8C was held for 2.50 min.
After that the oven temperature was
increased to 10 8C at 2.5 8C per min, increased to 80 8C at 10 8C per min, increased to 150 8C at 20 8C per min, and
then increased to 180 8C at 10 8C per
min and was held for 2 min at that
temperature. Constant column pressure at 20.5 psi was maintained. The
ionization potential of the mass selective detector (Model 5973; HewlettPackard Co.) was 70 eV, and the scan
range was 33.1 to 350 m/z. Identification of volatiles was achieved by comparing mass spectral data of samples
with those of the Wiley library
(Hewlett-Packard Co.). Standards,
when available, were used to confirm
the identification by the mass selective
detector. The area of each peak was integrated using ChemStation software
(Hewlett-Packard Co.), and the total
ion count 3 104 was reported as an indicator of volatiles generated from the
meat samples.
Statistical analysis
Four packages of ham and sausage
each were used to analyze volatile and
fatty acid composition. Data were analyzed as 8 replications using SAS software (SAS Institute Inc. 1989). The generalized linear model (GLM) procedure
was used to calculate the means and
standard errors.
Results & Discussion
T
HE FATTY ACIDS OF C HINESE J INHUA
ham and Cantonese sausage were
828 JOURNAL OF FOOD SCIENCE—Vol. 66, No. 6, 2001
very similar ( Table 1). Both products
contained about 61 to 62% of total unsaturated fatty acids and 36 to 37% total saturated fatty acids. Their major
fatty acids were octadecenoic acid,
hexadecenoic acid, and octadecanoic
acid, in descending order. However,
Cantonese sausage had 4-fold higher
TBARS than the Jinhua ham (Table 1).
The high TBARS value of Cantonese
sausage could be related to the grinding and fermentation steps and high fat
content in Cantonese sausage. The fat
content of Cantonese sausage was
31.73%, much higher than that of the
Jinhua ham (20.84%; Table 1). The color of TBARS samples from Cantonese
sausage was orange-red, not a typical
red color, indicating that the major
TBARS were not malonaldehyde but
rather other oxidation compounds including ethanol and aldehydes. Many
compounds can react with thiobarbituric acid (TBA) to produce chromatic
products. Kosugi and others (1987) reported that alkanals and alkenals reacted with TBA and produced yellow (455
nm) and orange (495 nm) pigments.
Supporting the interpretation, alkanals
detected in volatiles indicated that
large amounts of alkanals and alkenals
were formed in Cantonese sausage,
probably via the extensive degradation
of lipids during the fermentation of
Cantonese sausage. This also suggested
that the TBARS measurement, determined at 531 nm, underestimated the
real oxidation status of Jinhua ham.
Ethanol and ethyl esters of fatty acids were the dominant volatile components (76% of total volatiles) in Chinese Cantonese sausage ( Table 2). In
Cantonese sausage, a large amount of
Volatiles of Chinese Ham and Sausage . . .
Table 2—Volatiles in Chinese Cantonese sausages
Volatiles
Ion counts (×104)
Ethanol and methyl/ethyl ethers
Ethanol
Acetic acid ethyl ester
1-Propene, 3-ethoxy-2-methyl ester
Propanoic acid ethyl ester
Propanoic acid 2-methyl-ethyl ester
Butanoic acid 2-methyl ethyl ester
Butanoic acid 3-methyl ethyl ester
Pentanoic acid, ethyl ester
Hexanoic acid, ethyl ester
Peptanoic acid, ethyl ester
Total ethanol-related volatiles
Alkanes and alkenes
Hexane
Heptane
1-Octene
Octane
Total alkanes, alkenes
Aldehydes
2-Methyl propanal
3-Methyl butanal
2-Methyl butanal
Pentanal
Hexanal
Total aldehydes
Ethyl butanoate
Methyl cyclopentane
Chloroform
Benzene
2-Pentanone
1,1-Diethoxy ethane
4-Heptanone
Methyl benzene
4-Heptanone
Total other volatiles
Total volatiles
Percentage
Identification
133319 6 27109
33760 6 13781
515 6 153
2779 6 1073
3030 6 926
216 6 192
222 6 286
316 6 260
3440 6 1252
320 6 73
177917
57.38
14.53
0.22
1.20
1.30
0.09
0.10
0.14
1.48
0.14
76.58
MS/RT 2
MS/RT
MS
MS
MS
MS
MS
MS/RT
MS/RT
MS/RT
2677 6 728
1499 6 465
265 6 222
1143 6 505
5584
1.15
0.65
0.11
0.49
2.40
MS/R
MS/RT
MS/RT
MS
244 6 77
1107 6 327
449 6 120
663 6 347
1911 6 1774
4374
40674 6 19048
353 6 118
809 6 189
180 6 46
265 6 217
1170 6 930
513 6 101
345 6 145
156 6 45
44465
232337 6 43747
0.11
0.48
0.19
0.29
0.82
1.88
17.51
0.15
0.35
0.08
0.11
0.50
0.22
0.15
0.07
1.63
100.00
MS
MS/RT
MS/RT
MS/R
MS/RT
Other volatiles
MS/RT
MS
MS/RT
MS/RT
MS/RT
MS
MS/RT
MS
MS/RT
1 n=4. 2 MS: identified by mass spectrometry; RT: identified by retention time of standards.
China. Briefly, the temperature of the
ripening chamber is raised 4 8C per mo
for the first 6 mo to 26 8C, then the temperature is lowered 4 8C per mo for 4
mo. Large amounts of n-alkanes,
branched alkanes, aldehydes, and ketones were found in Jinhua ham, but
ethanol and its derivatives were not detected. The sources of n-alkanes were
not clear, but they were probably derived from lipids. 1-Pentene, 2-pentene,
1-octene, and 2-octene could be
formed from the degradation of unsaturated fatty acids by a similar mechanism as n-alkanes, and hams contained
large amounts of unsaturated fatty acids, as shown in Table 1. Many
branched alkenes also were detected in
Jinhua ham. This was in agreement with
findings by Garcia and others (1991)
who reported on the volatile components of Iberian dry-cured ham. It is
difficult to derive the mechanisms of
branched-alkene formation, but the
most probable pathway for the forma-
tion of branched alkenes in Jinhua ham
could be through the degradation and
intermolecular rearrangement of fatty
acids. This conclusion is similar to that
reported by Ruiz and others (1999) who
detected high amounts of n-alkanes
and branched-alkanes in dry-cured Iberian ham. Sabio and others (1998) also
showed that Iberian ham had a high
amount of medium-chain hydrocarbon
compounds, similar to our results with
Jinhua ham. However, the volatile components of Bayonne, light Italian country, and Parma ham were somewhat
different from the Jinhua ham (Sabio
and others 1998). The common characteristic processing method of Iberian
ham and Jinhua ham is a very long aging time. During this aging period, a
large number of alkanes should be
formed through various reactions including lipid oxidation. Numerous
branched aldehydes were also detected
in Jinhua ham. The branched aldehydes
might be produced by enzyme-cata-
Vol. 66, No. 6, 2001—JOURNAL OF FOOD SCIENCE
829
Food Chemistry and Toxicology
wine (approximately 8% of meat, ethanol content about 15%) is usually
added at the beginning of the sausagemaking process. Sausages are fermented and oven-dried at about 50 8C
for 3 days, with relative humidity of
about 70%. High amounts of ethanol
and ethyl esters indicated that ethanol
and its derivatives played a central
role in the formation of the characteristic flavor of Cantonese sausage.
Chen and others (1997) also reported
that wine is important for the flavor
development of Chinese-style sausage. The purpose of using a large
amount of wine in Cantonese sausage
is: to produce characteristic flavor
and to prevent the spoilage of sausage
during the fermentation process. Because no lactic acid starter culture is
added in Cantonese sausage manufacture, as in other fermented sausages,
bacteria would grow quickly if a large
amount of ethanol, which inhibits
bacterial growth, was not added. In
sausages fermented with lactic acid
bacteria, the strains of lactic acid bacteria influence the generation of volatile components substantially (Berdague and others 1993b). Loury (1972)
suggested that n-alkanes might come
from the oxidation of unsaturated lipids. About 30 volatile compounds
were detected in the Cantonese sausage. Hexanal has been suggested as
the oxidation product of linoleic acid,
and the detection of hexanal among
the volatiles indicated that there was a
certain degree of lipid oxidation in the
Cantonese sausage. This was in agreement with the TBARS values shown in
Table 1. Branched aldehydes could
come from the degradation of amino
acids by oxidative deamination-decarboxylation of amino acids via the
Strecker degradation (Garcia and others 1991). Other volatiles, such as octane, heptane, pentane, heptanone,
octane, and hexane, could be formed
via the degradation of unsaturated
fatty acids or amino acids. Endogenous enzymes in meat and enzymes
from microorganisms, and the degradation of sugar and its reaction with
amino acids or lipids during fermentation, also could have contributed to
the formation of these volatiles.
The volatiles in Jinhua ham are quite
different from those of the Cantonese
sausage (Table 3). Processing of Jinhua
ham involves a long ripening process
(10 mo) at a relative humidity of about
70 %. The temperature during the ripening periods changes along with the
climate changes in the Jinhua area of
Volatiles of Chinese Ham and Sausage . . .
Table 3—Volatiles in Chinese Jinhua ham1
Volatiles
Ion counts (3
3 104)
Percentage
Identification
0.08
30.44
1.75
7.42
11.70
13.37
1.12
0.24
0.86
66.97
MS/RT 2
MS/RT
MS
MS/RT
MS/RT
MS/RT
MS
MS
MS
Food Chemistry and Toxicology
n-Alkanes and alkenes
Butane
70 6 56
Pentane
25907 6 5341
2-Pentene
1490 6 478
Hexane
6312 6 1096
Heptane
9956 6 3442
Octane
11378 6 4217
1-Octene
951 6 325
1-Pentene
207 ± 122
2-Octene
735 6 246
Total alkanes and alkenes
57006
Branched alkanes and alkenes
Methyl cyclopentane
417 6 79
Methyl cyclohexane
196 6 50
2,5-Dimethyl hexane
297 6 95
2,4-Dimethyl hexane
752 6 230
2,3,4-Trimethyl pentane
1488 6 441
2,3,3-Trimethyl pentane
1320 6 377
2,3-Dimethyl hexane
456 6 137
3-Methyl heptane
407 6 116
3-Methyl-2-heptene
1797 6 555
Total branched alkanes and alkenes
7130
Aldehydes
2-Methyl propanal
2668 6 866
3-Methyl butanal
5304 6 2052
2-Methyl butanal
4225 6 1425
Pentanal
518 6 349
Hexanal
143 6 93
Total aldehydes
12715
Sulfur compounds
Methanethiol
117 6 36
Thiobis methane
122 6 102
Thiourea
59 6 20
2,3-Dimethyl disulfide
735 6 208
Total sulfur compounds
1033
Other volatiles
2-Methyl furan
212 6 62
Chloroform
2880 6 835
Benzene
127 6 81
2-Ethyl furan
413 6 100
Methyl benzene
388 6 96
1,2 Cyclohexanedione
2938 6 877
Ethyl benzene
59 6 31
1,4 Dimethyl benzene
73 6 39
Total other volatiles
7090
Total volatiles
85116 6 17873
0.49
0.23
0.35
0.88
1.75
1.55
0.54
0.48
2.11
8.38
MS
MS
MS
MS
M
MS
MS
MS
MS
3.13
6.23
4.96
0.61
0.17
15.11
MS
MS/RT
MS/RT
MS/RT
MS/RT
0.14
0.14
0.07
0.86
1.21
MS/RT
MS/RT
MS/RT
MS/RT
0.25
3.38
0.15
0.49
0.46
3.45
0.07
0.09
8.33
100.00
MS
MS/RT
MS/RT
MS
MS
MS
MS
MS
1n=4. 2 MS: identified by mass spectrometry; RT: identified by retention time of standards.
lyzed and other chemical reactions.
Jose and others (1993) reported that
muscle lipases and esterases in drycured ham are quite stable and active
even after 15 mo of ripening, indicating
that endogenous enzymes could have
contributed to the formation of volatiles in dry-cured ham. Hinrichsen and
Pedersen (1995) examined the relationship between microorganisms and volatiles formed in meat products and
suggested that microorganisms contributed to volatile formation, especially those related to amino acid catabolism. About 15% of total volatiles
in Jinhua ham were aldehydes. Because
Jinhua hams were exposed to air for
about 1 yr during the manufacturing
process, large amounts of lipid oxidation products can be expected. Ruiz
and others (1999) reported that the
content of 2- and 3-methylbutanals,
the degradation products of amino acids (leucine and isoleucine), were higher in aged hams. Aldehydes in drycured ham may be correlated to the
aromas of ham and cured meats
(Buscailhon and others 1994). Most of
the volatile compounds found in drycured hams could be produced by lipid autoxidation and amino acid degradation (Sabio and others 1998). Four
sulfur compounds were detected in
the volatiles of Jinhua ham ( Table 3).
Although the amounts of sulfur compounds were relatively small, they can
830 JOURNAL OF FOOD SCIENCE—Vol. 66, No. 6, 2001
be expected to play an important role
in the overall taste/odor of ham, because their threshold values are much
lower than those of other volatile
compounds.
Chloroform, benzene, and methyl
benzene were found in both Cantonese
sausage and Jinhua ham. These volatiles
were not from the meat products but
from the packaging materials. We observed the migration of these compounds from packaging materials to
raw and cooked meat during storage
(unpublished data).
Conclusion
T
HIS STUDY INDICATED THAT THE MA-
jority of volatiles from Cantonese
sausage are alcohol derivatives. Only a
small proportion of volatiles was generated from fatty acids and amino acids.
The volatiles in Jinhua ham, however,
were very different from those of the
Cantonese sausage in that most of the
volatile components were the breakdown products of fatty acids and amino
acids. The alcohol-derived volatiles in
Cantonese sausage should be related to
the large amount of wine added during
manufacturing, while the volatiles in
Jinhua ham would be related to the
long aging process.
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Journal Paper Nr. J - 19121 of the Iowa Agriculture and Home
Economics Experiment Station, Ames, IA 50011. Project Nr.
3322, supported by Hatch Act and State of Iowa funds.
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Dept., Iowa State Univ., Ames, IA 50011-3150.
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