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Analytical Methods
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Article history: The volatile flavour components of different teas growing in Thailand were extracted using the simulta-
Received 23 June 2009 neous distillation and extraction (SDE) technique. These volatiles were investigated by GC–MS. At least
Received in revised form 18 June 2010 54 components representing 76.51–83.32% of all samples were identified. Hotrienol, geraniol and linalool
Accepted 19 September 2010
were found to be the major components in Green Oolong tea. Green Assam tea contained linalool, gera-
niol and a-terpineol as the key flavour constituents. Chin Shin Oolong tea was dominated by linalool,
indole and cis-jasmone whilst the major flavour volatiles of Chin Hsuan Oolong tea were trans-nerolidol,
Keywords:
cis-jasmone and geraniol. Indole, geraniol and cis-jasmone were detected as the main constituents in Four
Camellia sinensis
Flavour volatiles
Season tea. Change of quality and quantity of volatile flavour components was related to fermentation
GC–MS methods that increased volatiles were illustrated by the semi-fermented tea processing method. Green
Antioxidant activity Assam tea infusion extract was evaluated to have the strongest antioxidant activities with the highest
Phenolic content amount of phenol content followed by Four Season tea, Chin Shin Oolong tea, Chin Hsuan Oolong tea
Tea and Green Oolong tea, respectively.
Simultaneous distillation and extraction Ó 2010 Elsevier Ltd. All rights reserved.
(SDE)
1. Introduction & Owuor, 1992; Sanderson & Graham, 1973; Takeo, 1981). The
quality and variation of volatile flavour components in tea is due
Tea (Camellia sinensis) is and has been one of the most popular to different environmental and ecological conditions and the tea
drinks in the world for over 4000 years. More than 3 million hect- processing method. Polyphenols are key non-volatile components
ares of the world has been used for planting tea (Ravichandran & which also play important role in tea. This component provides
Parthiban, 1998). Nowadays, tea is used in pharmaceutical and important beneficial effects of tea in term of the antioxidant activ-
industrial applications (Almajano, Carbó, Jiménez, & Gordon, ity and free radical scavenging ability (Frei & Higdon, 2003). Tea
2008; Falé et al., 2009; Ravichandran, 2002). Tea is manufactured contains predominantly polyphenols such as catechins, monomeric
in many areas over the world such as Japan, Taiwan, India as well flavonols, and ()-epigallocatechin gallate (EGCG) which are de-
as Thailand. Tea processing methods are classified by the degree of tected in fresh tea leaves at a high level (Almajano et al., 2008;
fermentation. Green tea is produced by a non-fermentation tea Chattopadhyay et al., 2004; Rietveld & Wiseman, 2003). The fer-
process whilst Oolong and red tea are established from partial fer- mentation process also affect to polyphenols in tea. Flavanols, flav-
mentation. A completely fermented process is applied to black tea. andiols and phenolic acids like gallic acid, cumaric acid and caffeic
The taste and flavour of tea is controlled by key chemical compo- acid are predominant in non-fermented green tea whilst some
nents which are volatile terpenes, caffeine, organic acids and poly- polyphenols are oxidised, degraded and polymerised enzymati-
phenols (Borse, Rao, Nagalakshmi, & Krishnamurthy, 2002). Tea cally to theaflavins and thearubigens in black tea during the full
volatiles can be divided into two groups comprising non-terpenoid fermentation tea processing method (Liebert, Licht, Böhm, & Bits-
and tepenoid components. Non-terpenoids found in tea are hexe- ch, 1999; Weisburger, 1996). Katalinic, Milos, Kulisic, and Jukic
nols providing fresh green flavour (Rawat et al., 2007) and products (2006) reported that tea variety and the content of EGCG con-
of lipid degradation (Ganeshan & Ramasamy, 1996; Mahanta, trolled the antioxidant effectiveness. High content of EGCG and
Tamuli, & Bhuyan, 1993; Robinson & Owuor, 1992). The detected ()-epigallocatechin (EGC) was detected in non-fermented and
terpenoid components in tea are monoterpene alcohols, including semi-fermented tea, e.g. green and Oolong tea. Conversely, the
linalool and geraniol, which impart a sweet floral aroma (Robinson content of both components were much lower in black tea.
The purpose of this present study was to investigate the finger-
⇑ Corresponding author. Tel.: +66 53 9166788; fax: +66 53 916776. print of volatile flavour components of Thai teas obtained from
E-mail address: patcharee_pri@mfu.ac.th (P. Pripdeevech). Mae Salong Mountain, Chiang Rai province, in the northern part
0308-8146/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.09.074
798 P. Pripdeevech, T. Machan / Food Chemistry 125 (2011) 797–802
of Thailand which is the most important area for planting tea in employed during the brewing process. After 10 min, the tea infu-
Thailand. The different tea infusions extracts were evaluated for sion was filtered under vacuum and cooled to room temperature.
their antioxidant activity and phenolic content. The obtained tea extract was concentrated using a vacuum rotary
evaporator and dried to constant weight in an oven at 105 °C.
2. Materials and methods
2.3.2. DPPH radical scavenging assay
2.1. Tea samples The effect of tea extracts on the content of 2,2-diphenyl-2-pic-
rylhydrazyl radical (DPPH) were evaluated by a spectrophotomet-
Five tea samples manufactured under Thai Tea Suwirun ric method based on the reduction of a methanol solution of DPPH
Partnership, Mae Salong Mountain, Chiang Rai, Thailand were used according to the modified method of Blois (1958). One milliliter of
in this study. These five different tea samples with different qual- various concentrations of the each sample in methanol was added
ities consisted of non-fermentation tea (Green Oolong tea and to 1 mL of a 0.003% methanol solution of DPPH and the reaction
Green Assam tea) as well as semi-fermentation tea (Chin Shin mixture was shaken vigorously. The tubes were allowed to stand
Oolong tea, Chin Hsuan Oolong tea and Four Season tea). Each at room temperature (27 °C) for 30 min. Each reaction mixture
sample lacked any additional manipulation and was kept under a was then placed in the cuvette holder of the Perkin–Elmer-Lamda
temperature of 5 °C before being subjected to simultaneous distil- 25 UV/vis spectrophotometer and monitored at 517 nm against a
lation–extraction (SDE). All solvents were of analytical grade, blank which used methanol as the baseline correction. The scav-
dichloromethane, anhydrous sodium sulphate, mixtures of C8–C22 enging ability was calculated as follows: scavenging ability
n-alkanes and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and gallic (%) = 100 [Absorbance of control–Absorbance of sample/Absor-
acid were purchased from Merck (Darmstadt, Germany). bance of control]. The antioxidant activity of all tea extracts was
expressed as IC50 which was defined as the concentration (in lg/
2.2. Analysis of volatile flavour constituents mL) of tea extract required to inhibit the formation of DPPH radi-
cals by 50%. The experiment was carried out in triplicate and the
2.2.1. Extraction results are the mean values.
The extraction was carried out in a modified Likens–Nickerson
SDE apparatus for 5 h. Two hundred grams of tea samples and
2.4. Determination of total phenolic contents
200 mL of distilled water were added to a 500 mL round-bottom
flask. Dichloromethane (150 mL) was added to another 250 mL
Total phenolic content of tea extracts obtained from different
round-bottom flask. Both flasks were connected to the apparatus,
samples was determined using the Folin–Ciocalteu reagent accord-
and more dichloromethane and distilled water were added into
ing to the modified method of Singleton and Rossi (1965) using
the central arm of the apparatus. The flask containing the dichloro-
gallic acid as standard. The tea solution (0.2 mL) was mixed with
methane was heated by using a water bath at 50 °C, whilst the flask
1.0 mL of Folin–Ciocalteu reagent, 1.0 mL of an aqueous solution
containing the tea leaves and distilled water was heated by using a
of 7% Na2CO3 and 5.0 mL of distillated water, respectively. Then,
paraffin oil bath at 200 °C. After extraction, the distillate in a con-
the mixture was vortexed vigorously. The reaction mixtures were
ical flask was concentrated using a vacuum rotary evaporator and
allowed to stand for 30 min before absorbance at 765 nm was mea-
then all sample essential oils were stored in the dark at 4 °C. All tea
sured. The same procedure was also applied to the standard solu-
essential oil samples obtained were diluted to 1:10 v/v with
tions of gallic acid. The calibration equation for gallic acid was
dichloromethane prior to injection into the GC–MS instruments.
y = 0.00515x 0.00400 (R2 = 0.999) where y is the absorbance
and x is the concentration of gallic acid in mg/mL.
2.2.2. Gas chromatography–mass spectrometry (GC–MS)
The volatile flavour constituents of each sample were analysed
using an HP model 6890 gas chromatograph equipped with an 3. Results and discussion
HP-5MS (5% phenyl-polymethylsiloxane) capillary column
(30 m 0.25 mm i.d., film thickness 0.25 lm; Agilent Technologies, 3.1. Volatile flavour compounds analysis
USA) interfaced to an HP model 5973 mass-selective detector. The
oven temperature was initially held at 60 °C and then increased The low percentage yield (0.01–0.03% w/w) of tea essential oils
by 2 °C/min to 250 °C. The injector and detector temperatures were was obtained from all tea samples extracted by SDE technique. Re-
250 and 280 °C, respectively. Purified helium was used as the car- sults of the fingerprint in terms of volatile flavour compounds of all
rier gas at a flow rate 1 mL/min. EI mass spectra were collected at tea essential oils analysed by GC–MS and their profiles as well as
70 eV ionisation voltages over the range of m/z 29–300. The elec- quantities determined with the reference to internal standard in
tron multiplier voltage was 1150 V. The ion source and quadrupole different tea samples are summarised in Fig. 1 and Table 1. Sixty-
temperatures were set at 230 and 150 °C, respectively. Identifica- three volatiles were identified representing 81.37% of the essential
tion of the volatile components was performed by comparison of oil obtained from Green Oolong tea. Monoterpene alcohols of hot-
their Kovát retention indices, relative to C8–C22 n-alkanes, and com- rienol (15.27%), geraniol (10.88%), linalool (10.71%) and a-terpineol
parison of the mass spectra of individual components with the ref- (5.78%) were found to be the major components. Small amounts
erence mass spectra in the Wiley 275, NIST 98 databases and Adams were coumaran (3.14%), nerol (1.82%) and indole (1.74%) were also
1989 with corresponding data of volatile flavour components in tea detected. Fifty-four components representing 83.32% in Green As-
(Bastos, Ishimotoa, Marquesb, Ferrib, & Torres, 2006; Borse et al., sam tea essential oil were identified. Linalool (15.61%), geraniol
2002; Kumazawa & Masuda, 2002; Ravichandran & Parthiban, (14.22%), a-terpineol (11.27%) and nerol (5.26%) were found to
1998; Rawat et al., 2007; Wang et al., 2008). be the key flavour constituents in Green Assam tea whilst benzyl
ethanoate (3.73%), phytol (3.37%) and trans-nerolidol (2.41%) were
2.3. Antioxidant activity minor components. A total of 60 constituents representing 81.85%
of the Chin Shin Oolong tea essential oil were demonstrated. The
2.3.1. Preparation of tea extract dominant components were linalool (14.33%), indole (13.86%),
Each extract was prepared by adding 100 mL of boiling distilled cis-jasmone (12.15%) and trans-nerolidol (7.35%). They were
water to 10 g of tea sample. Stirring using a magnetic stirrer was accompanied by the small amounts of hotrienol, a-terpineol,
P. Pripdeevech, T. Machan / Food Chemistry 125 (2011) 797–802 799
Table 1
Volatile flavour compounds of all tea essential oils.a
Table 1 (continued)
it can be said that Four Season tea was evaluated to be the impor-
tant tea having the best quality of aroma and antioxidant ability.
4. Conclusion
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