Variations in the components of Osmanthus fragrans Lour.

essential oil
at different stages of owering
Li-mei Wang, Mao-teng Li
*
, Wen-wen Jin, Shuo Li, Shuai-qi Zhang, Long-jiang Yu
*
Institute of Resource Biology and Biotechnology, College of Life Science and Technology, Huazhong University of Science and Technology, Luoyu Road 1037, Wuhan 430074, China
a r t i c l e i n f o
Article history:
Received 20 June 2008
Received in revised form 29 July 2008
Accepted 16 September 2008
Keywords:
Osmanthus fragrans Lour.
Headspace solid-phase microextraction
Gas chromatographymass spectrometry
Chemical components
a b s t r a c t
A headspace solid-phase microextraction (HS-SPME) method was used to extract the essential oil of
Osmanthus fragrans Lour., which was then analysed by gas chromatographymass spectrometry (GC
MS) at four different stages of owering. The primary chemical components of the essential oil extracted
from O. fragrans were linalool and its oxide, a-ionone, b-ionone, nerol, c-decalactone, 9,12,15-octadec-
atrienoic acid, and hexadecanoic acid, most of which were at their highest concentrations in extracts
obtained from owers at the initial owering stage. HS-SPME using a PDMS/DVB bre is a simple, rapid,
and solvent-free method for the extraction of volatile compounds emitted from living O. fragrans. HS-
SPME with GCMS can be used to determine the aromatic maturity in O. fragrans, to decide the optimal
harvest date. The initial owering stage was the best time to harvest O. fragrans.
2008 Elsevier Ltd. All rights reserved.
1. Introduction
Osmanthus fragrans Lour. is a plant belonging to the Oleaceae
family (mura, Honda, & Hayashi, 2000), which is widely distrib-
uted in the south and middle of China. Osmanthus fragrans is con-
sidered one of the four famous traditional owers of China and is
widely cultivated as an ornamental plant today (Wang, Pan, Tang,
& Huang, 2006). Five cities in China (Hangzhou, Suzhou, Guilin,
Chengdu, and Xianning) comprise the production centre for O. frag-
rans (Zang, Xiang, Liu, & Hao, 2003). In China, O. fragrans has been
studied since the 1940s (Huang & Huang, 1949) and plays an
important role in Chinese ower culture. The ower of O. fragrans
is of economic importance because it is used as an additive for
foods, tea, and other beverages (Yang, Yao, & Qian, 2005; Yang &
Zhu, 2000). The concrete and essential oil of O. fragrans are consid-
ered as one of the best natural essences, which are only used in the
most expensive perfumes and cosmetics (Jin, Zheng, Jin, Chen, &
Wang, 2006). Research into O. fragrans has mainly related to its
germplasm resources, classication, tissue culture, cultivation,
propagation, the physiological hardiness of the plants, its chemical
composition, and the senescence of its owers (Liu & Xiang, 2003;
Qiu, Hu, Chen, Chen, & Wu, 2004; Song, Ge, & Wang, 2005; Zhu &
Zeng, 2000). During its long evolution, under the effects of natural
and articial selection, many varieties of O. fragrans have devel-
oped and are divided into the Semperoren, Thunbergii, Latifolius,
and Aurantiaeus groups (Zang et al., 2003). Research into the
essential oil of O. fragrans has demonstrated that its composition
is very different at different stages of Osmanthus ower develop-
ment (Yang & Zhu, 2000).
Until now, few studies have reported the analysis of the essen-
tial oil at different stages of Osmanthus owering. In this work, we
propose a method for the rapid analysis of volatile compounds re-
leased at four different stages from living Osmanthus owers, using
headspace solid-phase microextraction (HS-SPME) and gas chro-
matographymass spectrometry (GCMS). In this study, we report
the variation of chemical composition of different owering stages,
in order to determine the optimal harvest time for O. fragrans.
2. Experimental
2.1. Plant material
Fresh owers of O. fragrans were collected on the campus of
Huazhong University of Science and Technology, Wuhan, China.
The owering period of O. fragrans lasts for 710 days. According
to previous studies, the owering stage can be divided into four
periods, i.e., the xiangyan stage (A), the initial owering stage
(B), the full owering stage (C), and the nal owering stage (D)
(Yang & Zhu, 2000).
2.2. Extraction of O. fragrans concrete and essential oil
Osmanthus fragrans concrete was extracted by the traditional
method using petroleum ether (Mai, 2000). The essential oil was
extracted with anhydrous alcohol according to the method of the
0308-8146/$ - see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2008.09.044
* Corresponding authors. Tel./fax: +86 (0)27 87792265.
E-mail addresses: limaoteng426@163.com (M.-t. Li), yulj@mail.hust.edu.cn (L.-j.
Yu).
Food Chemistry 114 (2009) 233236
Contents lists available at ScienceDirect
Food Chemistry
j our nal homepage: www. el sevi er . com/ l ocat e/ f oodchem
National Standards of the Peoples Republic of China GB 6780-98
(Food Additive Osmanthus oncrete).
2.3. Headspace solid-phase microextraction of volatile compounds
A headspace SPME holder, serum bottles, glass liners and
poly(dimethylsiloxane)/divinylbenzene (PDMS/DVB) bres of
65 lm were purchased from Supelco (Bellefonte, PA). One 12-
year-old latifolius tree, which owered from September 18 to 27,
2007, was selected for the experiment. HS-SPME was used to ex-
tract the volatile compounds during the four different develop-
mental owering stages. A branch of O. fragrans with about 100
owers was placed into the sampling chamber. Fibres of PDMS/
DVB were applied to extract the volatile compounds emitted by
the O. fragrans owers. The HS sampling technique was as follows.
Sample vials, which contained four owers, were placed into a
water bath at 50 C for 40 min to allow partition equilibration of
the sample and HS. The analytes adsorbed by the bres were des-
orbed at 250 C for 3 min and analysed by GCMS (Deng, Song, &
Hu, 2004).
2.4. GCMS analysis
Desorption analysis was performed on an Agilent 7890 GC
system (Agilent, Santa Clara, CA) coupled to an Agilent 5975C
quadrupole mass spectrometer. The analytes were separated using
an HP-5MS capillary column (30 m 0.25 mm 0.25 m), which
was inserted directly into the ion source of the MS system. The
oven temperature was programmed as follows: 50 C for 2 min,
which was increased to 100 C at 10 C/min, then increased to
250 C at 5 C/min, where it was maintained for 5 min. Helium
(99.999%), which was used as the carrier, was provided at a ow
rate of 1.5 ml/min. The split ratio was 50:1. The detector tempera-
ture was set at 250 C. The electron impact ionisation conditions
were an ion energy of 70 eV and the mass range scanned was
50450 amu in the full-scan acquisition mode. Each compound
was identied using the Wiley/NBS, NIST, and other published
mass spectral libraries of GCMS data (Adams, 2001; Msaada
et al., 2007).
3. Results and discussion
3.1. Comparison of different extraction methods
Traditionally, volatile compounds are frequently extracted by
hydrodistillation, supercritical carbon dioxide extraction, or sol-
vent extraction (Bakkali, Averbeck, Averbeck, & Idaomar, 2008;
Liu, He, & Shen, 1996). The hydrodistillation and supercritical car-
bon dioxide extraction methods are time-consuming, and large
amounts of solvents are required in solvent extraction methods.
HS-SPME with GCMS is considered a rapid, simple, sensitive,
and solvent-free method for volatile compound analysis in plants.
SPME using a PDMS/DVB bre is considered the best method for
the extraction of volatile compounds (Shang, Hua, Deng, & Hua,
2002; Snchez-Palomo, Daz-Maroto, & Prez-Coello, 2005). In this
work, ower emissions of O. fragrans were extracted at different
owering stages using SPME with PDMS/DVB bre and then anal-
ysis by GCMS. Most of the compounds in the Osmanthus ower
fragrance were separated and identied, and some of them had
not been found before. About 46 h is required for traditional
methods, whereas only 40 min was needed for the HS-SPME meth-
od. Furthermore, hundreds of owers are necessary for ower
emission extraction by traditional methods, whereas only 46
owers were sufcient for compound analysis in this study. This
method is suitable for the extraction of volatile compounds from
living owers and reects the natural release of aromas with great-
er delity.
3.2. Yield and components of the essential oil
The chemical composition of the O. fragrans essential oil was
identied with a mass spectra library and 24, 24, 29, and 22 com-
pounds were identied in the xiangyan, initial, full, and nal ow-
ering stages, respectively (Table 1), which accounted for 96.01%,
97.27%, 96.78%, and 95.25%, respectively, of the essential oil ex-
tracted from the O. fragrans owers.
The sunlight that plants receive may inuence the compounds
that accumulate in a particular period in response to environmen-
tal changes (Koenen, 2001). Research has shown that a variety of
geographical and ecological factors lead to quantitative and
qualitative differences in essential oils (Argyropoulou, Daferera,
Tarantilis, Fasseas, & Polyssiou, 2007; Santos-Gomes, Fernandes-
Ferreira, & Vicente, 2005). Our experimental results show that
the amount of concrete production of O. fragrans during various
developmental stages is as follows: the full owering stage > the
initial owering stage > the nal owering stage > the xiangyan
stage (Fig. 1). The essential oil content of O. fragrans was as follows:
the initial owering stage > the full owering stage > the nal
owering stage > the xiangyan stage (Fig. 2). Fig. 1 shows yields
of 0.174%, 0.272%, 0.281%, and 0.194% (w/w) of concrete from
100 g of Osmanthus owers by solvent extraction in the xiangyan,
initial, full, and nal owering stages, respectively, based on fresh
weights. Further analysis revealed that the content of essential oil
in the concrete during the different developmental stages was
65.7%, 80.5%, 64.4%, and 54.3%, respectively. Based on these results,
we calculated that the maximum essential oil production occurred
in the initial owering stage (2.868 mg per single ower), which
has rarely been reported in other papers.
The composition of the essential oil is affected by many factors,
such as the developmental stage, the extraction method, and the
conditions of analysis (Kim & Lee, 2004; Sangwan, Farooqi, Shabih,
& Sangwan, 2001). The chemical compositions of the various ex-
tracted essential oils are presented in Table 1. The major compo-
nents in the different owering stages of O. fragrans were
monoterpene alcohols, monoterpene ketones, fatty acids and their
esters (except in the xiangyan owering stage): linalool, trans-lin-
alool oxide, cis-linalool oxide, a-ionone, b-ionone, nerol, c-decalac-
tone, hexadecanoic acid, 9,12,15-octadecatrienoic acid,
hexadecanoic acid ethyl ester, and 9,12,15-octadecatrienoic acid
methyl ester (Table 1). Most of these constituents reached their
highest concentrations in the initial developmental stage (linalool
2.18%, nerol 3.27%, c-decalactone 5.09%, b-ionone 19.51%, 4-hydro-
xy-b-ionone 6.14%, hexadecanoic acid ethyl ester 4.13%, 9,12,15-
octadecatrienoic acid 16.93%, and 9,12,15-octadecatrienoic acid
methyl ester 6.20%) (Table 1). trans-Linalool oxide (13.23%)
reached its highest concentration at the full owering stage,
whereas cis-linalool oxide (25.70%), hexadecanoic acid (15.09%),
and 3-hydroxybutyl-2,2,4-trimethyl-2-cyclohexen-1-one (22.13%)
peaked at the end of the owering stage. The primary constituents
of the essential oil extracted during the xiangyan stage were
a-ionone (11.67%), c-decalactone (6.79%), b-ionone (5.95%),
5,6,7,7-tetrahydro-4,4,7-trimethyl-2(4H)-benzofuranone (29.73%),
9,12,15-octadecatrienoic acid (14.50%), and hexadecanoic acid
(10.56%) (Table 1). Other compounds identied in the sample
extracts are listed in Table 1.
The composition of the essential oil of O. fragrans extracted by
the hydrodistillation, supercritical carbon dioxide, and solvent
methods varies greatly. The primary components that we identi-
ed in this study were consistent with the results of most studies.
b-Ionone (12.59%) and 2-hydroxy-b-ionone (11.72%) (Liu et al.,
1996) were the primary constituents of the essential oil extracted
234 L.-m. Wang et al. / Food Chemistry 114 (2009) 233236
with supercritical carbon dioxide; b-ionone (10.66%), hexadecanoic
acid (11.74%), and linolenic acid methyl ester (9.04%) were the pri-
mary constituents of the essential oil extracted with solvent (Mai,
2000); and 3-methyl-2-butanol (13.26%), dimethyl phthalate
(15.13), butyl benzyl phthalate (21.91%), and alcohol mono-phthal-
ate (26.51%) were the primary constituents of the essential oil
extracted with hydrodistillation in O. fragrans (Xu & Lu, 2007). Di-
methyl phthalate, which was not likely to be a natural product
from the plant, was also observed in this study, which was also re-
ported by several researchers in other plants (Zeng et al., 2007;
Zhao et al., 2005, 2006).
In this study, we investigated the chemical composition of the
essential oil isolated from O. fragrans during different developmen-
tal stages, which has not been reported in other research. The most
signicant differences between the four developmental stages
were the changes in the percentages of linalool, trans-linalool
Table 1
The chemical composition of O. fragrans essential oil during four different developmental stages
Compounds Molecular weight Content (%) Molecular formula
A
a
B
b
C
c
D
d
Octane 114 0.64 0.08 C
8
H
18
5-Octen-1-ol 128 0.06 0.04 C
8
H
16
O
trans-Linalool oxide (furan) 170 0.75 2.02 12.35 4.03 C
10
H
18
O
2
cis-Linalool oxide (furan) 170 0.54 3.61 4.62 6.21 C
10
H
18
O
2
Linalool 154 0.32 2.18 0.03 C
10
H
18
O
Nonanal 142 0.35 2.43 C
9
H
18
O
2-Methylhexanoic acid 144 0.45 C
8
H
16
O
2
trans-Linalool oxide (pyran) 170 0.52 0.51 0.88 2.22 C
10
H
18
O
2
cis-Linalool oxide (pyran) 170 0.92 3.23 3.90 19.49 C
10
H
18
O
2
Lilac alcohol 170 2.70 2.73 C
10
H
18
O
2
Decanal 156 0.26 0.13 C
10
H
20
O
b-Ionol 194 0.21 0.07 C
13
H
22
O
Nerol 154 3.27 0.12 C
10
H
18
O
10-Pentadecen-1-ol 226 0.53 0.96 C
15
H
30
O
a-Ionone 192 11.67 3.26 0.27 0.12 C
13
H
20
O
Dihydro-b-ionone 194 0.77 0.37 0.08 C
13
H
22
O
c-Decalactone 170 6.79 8.09 1.16 C
10
H
18
O
2
b-Ionone 192 5.95 19.51 4.03 0.31 C
13
H
20
O
5,6,7,7-Tetrahydro-4,4,7-trimethyl-2(4H)-benzofuranone 208 29.73 0.90 0.94 C
13
H
20
O
2
4-Hydroxybenzeneethanol 138 5.16 C
8
H
10
O
2
Hexadecane 226 2.41 C
16
H
34
4-Hydroxy-b-ionone 208 3.99 8.14 2.74 4.25 C
13
H
20
O
2
3-Oxo-b-ionone 206 0.12 0.21 1.22 C
13
H
18
O
2
3-(3-Hydroxybutyl)-2,2,4-trimethyl-2-cyclohexen-1-one 210 0.05 8.07 22.13 C
13
H
22
O
2
9-Octadecenoic acid 282 1.45 0.23 0.41 1.69 C
18
H
34
O
2
2,6,10-Trimethyltetradecane 240 0.17 0.21 3.58 C
17
H
36
6,10,14-Trimethyl-2-pentadecanone 268 0.24 0.25 0.11 0.15 C
18
H
36
O
Dibutyl phthalate 278 0.59 0.80 0.62 1.92 C
16
H
22
O
4
Hexadecanoic acid 256 9.56 12.63 7.21 15.09 C
16
H
32
O
2
Hexadecanoic acid ethyl ester 284 1.25 4.13 0.50 C
18
H
36
O
2
Octadecanal 268 0.61 0.74 2.03 3.37 C
18
H
36
O
9,12,15-Octadecatrienoic acid 280 14.50 16.93 7.28 5.56 C
18
H
32
O
2
9,12,15-Octadecatrienoic acid, methyl ester 292 3.12 4.20 4.88 1.19 C
19
H
32
O
2
Docosane 310 8.51 C
22
H
46
1-Docosanol 326 11.68 C
22
H
46
O
Sum 94.89 97.27 96.78 95.25
Note:
a,b,c,d
represent the xiangyan stage, the initial owering stage, the full owering stage, the nal owering stage, respectively.
0
0.5
1
1.5
2
2.5
3
3.5
A B C D
Different flowering stage
T
h
e

y
i
e
l
d

o
f

O
.
f
r
a
g
r
a
n
s

c
o
n
c
r
e
t
e
(
m
g
/
g

F
W
)
0
2
4
6
8
10
12
14
16
18
W
e
i
g
h
t

o
f

p
e
r

s
i
n
g
l
e

f
l
o
w
e
r
(
m
g
.

F
W
)
yield of concrete
weight of per single flower
Fig. 1. The yield of O. fragrans concrete during different owering stages. Note: A, B,
C, D represents the xiangyan stage, the initial owering stage, the full owering
stage, the nal owering stage, respectively.
0
0. 1
0. 2
0. 3
0. 4
0. 5
0. 6
0. 7
0. 8
0. 9
A B
C D
Different flowering stage
T
h
e

c
o
n
t
e
n
t

o
f

e
s
s
e
n
t
i
a
l

o
i
l

o
f
O
.
f
r
a
g
r
a
n
s

c
o
n
c
r
e
t
e

(
w
/
w
)
Fig. 2. The content of essential oil of O. fragrans concrete during different owering
stages. Note: A, B, C, D represents the xiangyan stage, the initial owering stage, the
full owering stage, the nal owering stage, respectively.
L.-m. Wang et al. / Food Chemistry 114 (2009) 233236 235
oxide, cis-linalool oxide, a-ionone, b-ionone, nerol, c-decalactone,
hexadecanoic acid, 9,12,15-octadecatrienoic acid, hexadecanoic
acid, ethyl ester, and 9,12,15-octadecatrienoic acid methyl ester.
According to our results, the best time to harvest the owers for
their essential oil is in the initial owering stage.
4. Conclusion
This HS-SPME technique using a PDMS/DVB bre is a fast and
useful method for the quantication of the volatile compounds
emitted from living Osmanthus owers during different develop-
mental stages. It was revealed here that the chemical composition
differed signicantly during the maturation process, suggesting
that these differences reect changes in secondary metabolism.
We consider that the best harvesting time for the essential oil of
O. fragrans is the initial owering stage. This method can also be
used to monitor and rapidly predict the quality of the essential
oil from O. fragrans to decide the optimal harvest date. This method
can possibly be extended to the analysis of other aromatic plants.
Acknowledgements
This investigation was supported by the National Technology
Support Program (No. 2006BAC01A16). The authors are grateful
to Ms Cheng of Analytical and Testing Centre of Huazhong Univer-
sity of Science and Technology for their valuable GCMS analysis.
References
Adams, R. P. (2001). Identication of essential oil components by gas chromatography/
mass spectroscopy. USA: Allured Publishing Corporation, Carol Stream Illinois.
Argyropoulou, C., Daferera, D., Tarantilis, P., Fasseas, C., & Polyssiou, M. (2007).
Chemical composition and variation during development of the essential oil
from leaves of Lippia citriodora H.B.K. (Verbenaceae). Biochemical Systematics
and Ecology, 35, 831837.
Bakkali, F., Averbeck, S., Averbeck, D., & Idaomar, M. (2008). Biological effects of
essential oils. Food and Chemical Toxicology, 46, 446475.
Deng, C. H., Song, G. X., & Hu, Y. M. (2004). Application of HS-SPME and GCMS to
characterization of volatile compounds emitted from Osmanthus owers. Annali
di Chimica, 94, 921927.
Huang, Y. Y., & Huang, D. L. (1949). Theories of owers. Shanghai: New Century Press.
Jin, H. X., Zheng, H., Jin, Y. J., Chen, J. Y., & Wang, Y. (2006). Research on major
volatile components of 4 Osmanthus fragrance cultivars in Hangzhou Maolong
Guiyu Park. Forest Research, 19, 612615.
Kim, N. S., & Lee, D. S. (2004). Headspace solid-phase microextraction for
characterization of fragrances of lemon verbena (Aloysia triphylla) by gas
chromatographyemass spectrometry. Journal of Separation Science, 27, 96100.
Koenen, E. V. (2001). Medicinal poisonous and edible plants in Namibia. Windhoek:
Klaus Hess Publishers.
Liu, H., He, Z. Y., & Shen, M. Y. (1996). Study on the chemical constituents of absolute
oil from Osmanthus fragrans extracted by supercritical carbon dioxide.
Huangzhou Forestry Science, 25, 127131.
Liu, L. C., & Xiang, Q. H. (2003). Research progress on Osmanthus genus. Journal of
Nanjing Forestry University, 27, 8488.
Mai, Q. J. (2000). Study on chemical constituents of the absolute oil from ower of
Osmanthus fragrans. Journal of Guizhou University of Technology, 17, 7375.
Msaada, K., Hosni, K., Taarit, M. B., Chahed, T., Kchouk, M. E., & Marzouk, B. (2007).
Changes on essential oil composition of coriander (Coriandrum sativum L.) fruits
during three stages of maturity. Food Chemistry, 102, 11311134.
mura, H., Honda, K., & Hayashi, N. (2000). Floral scent of Osmanthus fragrans
discourages foraging behavior of cabbage buttery Pieris rapae. Journal of
Chemical Ecolog, 26, 655666.
Qiu, Y. X., Hu, S. Q., Chen, Y. L., Chen, X., & Wu, G. H. (2004). Studies on cultivar
classication of Osmanthus fragrans by ISSR-PCR analysis. Acta Horticulturae
Sinica, 31, 529532.
Snchez-Palomo, E., Daz-Maroto, M. C., & Prez-Coello, M. S. (2005). Rapid
determination of volatile compounds in grapes by HS-SPME coupled with
GCMS. Talanta, 66, 11521157.
Sangwan, N. S., Farooqi, A. H. A., Shabih, F., & Sangwan, R. S. (2001). Regulation of
essential oil production in plants. Plant Growth Regulation, 34, 321.
Santos-Gomes, P. C., Fernandes-Ferreira, M., & Vicente, A. M. S. (2005). Composition
of the essential oils from owers and leaves of Vervain (Aloysia triphylla
(LHerit.) Britton) grown in Portugal. Journal of Essential Oil Research, 17, 7378.
Shang, C. Q., Hua, Y. M., Deng, C. H., & Hua, K. J. (2002). Rapid determination of
volatile constituents of Michelia alba owers by gas chromatographymass
spectrometry with solid-phase microextraction. Journal of Chromatography A,
942, 283288.
Song, G. F., Ge, H., & Wang, C. Y. (2005). Primary study on in vitro culture and
micropropagation of sweet Osmanthus. Acta Horticulturae Sinica, 32, 738740.
Wang, H. S., Pan, Y. M., Tang, X. J., & Huang, Z. Q. (2006). Isolation and
characterization of melanin from Osmanthus fragrans seeds. LWT, 39, 496502.
Xu, J. M., & Lu, J. S. (2007). A study on chemical composition of the essential oils
from Osmanthus fragrant owers. Chinese Journal of Analysis Laboratory, 26,
3741.
Yang, K. M., & Zhu, W. J. (2000). Osmanthus fragrans. Shanghai Press of Science and
Technology.
Yang, W. P., Yao, W. R., & Qian, H. (2005). Effect of b-D-glucosidase on aromatic
compounds of sweet Osmanthus. Fine Chem Icals, 22, 924926.
Zang, D. K., Xiang, Q. B., Liu, Y. L., & Hao, R. M. (2003). The studying history and the
application to international cultivar registration authority of sweet Osmanthus
(Osmanthus fragrans Lour). Journal of Plant Resource and Environment, 12, 4953.
Zhu, C., & Zeng, G. W. (2000). Physiological and biochemical changes in ower
senescence of Osmanthus fragrans Lour.. Acta Horticulturae Sinica, 27, 356360.
Zeng, Y. X., Zhao, C. X., Liang, Y. Z., Yang, H., Fang, H. Z., Yi, L. Z., et al. (2007).
Comparative analysis of volatile components from Clematis species growing in
China. Analytica Chimica Acta, 595, 328339.
Zhao, C. X., Li, X. N., Liang, Y. Z., Fang, H. Z., Huang, L. F., & Guo, F. Q. (2006).
Comparative analysis of chemical components of essential oils from different
samples of Rhododendron with the help of chemometrics methods.
Chemometrics and Intelligent Laboratory Systems, 82, 218228.
Zhao, C. X., Liang, Y. Z., Fang, H. Z., & Li, X. N. (2005). Temperature-programmed
retention indices for gas chromatographymass spectroscopy analysis of plant
essential oils. Journal Chromatography A, 1096, 7685.
236 L.-m. Wang et al. / Food Chemistry 114 (2009) 233236