Short communication

Antioxidant properties of tropical and temperate herbal teas

E.W.C. Chan, Y.Y. Lim*, K.L. Chong, J.B.L. Tan, S.K. Wong
School of Science, Monash University Sunway Campus, Bandar Sunway, 46150 Petaling Jaya, Selangor, Malaysia
1. Introduction
For centuries, eastern countries have been using herbal
remedies to treat infections, ailments and diseases. Herbal
remedies are often consumed in the form of tea, i.e. an infusion
of dried plant parts steeped in boiling water. Herbal teas have been
gaining popularity in western countries in recent years (Manteiga
et al., 1997). Hundreds of different herbal teas are sold in health
food stores. Available as pure or blended samples, herbal teas are
popular because of their fragrance, antioxidant properties and
therapeutic applications (Naithani et al., 2006; Aoshima et al.,
2007). Tea from Camellia sinensis is the most widely consumed
beverage in the world, second only to water (Muktar and Ahmad,
2000). It is an important dietary source of natural phenolic
antioxidants (Lachman et al., 2003; Dimitrios, 2006).
Extensive research has been carried out on the antioxidant
properties (AOP) of green and black C. sinensis teas (Manzocco et
al., 1998; Dufresne and Farnworth, 2001). AOP of herbal teas of
temperate plants, mainly of Lamiaceae, have been well studied
(Triantaphyllou et al., 2001; Atoui et al., 2005; Capecka et al.,
2005). AOP of tropical herbal teas have been studied less, with
some analyses carried out on Aspalathus linearis (von Gadowet al.,
1997; Erickson, 2003); Cymbopogon citratus (Tsai et al., 2007;
Aoshima et al., 2007); Hibiscus sabdariffa (Aoshima et al., 2007);
Phyllanthus amarus (Limand Murtijaya, 2007); Psidiumguajava and
Toona sinensis (Chen et al., 2007); and Thunbergia laurifolia and
Orthosiphon aristatus (Chan and Lim, 2006).
In this study, AOP of thirteen tropical and ve temperate herbal
teas were screened, with comparisons with green, oolong and
black teas of C. sinensis carried out as positive controls. The AOP
studied were total phenolic content, ascorbic acid equivalent
antioxidant capacity (AEAC), ferric-reducing power (FRP) and
ferrous ion-chelating (FIC) ability. This study, to the best of our
knowledge, represents the most comprehensive comparison
between AOP of tropical and temperate herbal teas in the
literature.
2. Materials and methods
2.1. Herbal and Camellia teas
Herbal teas, together with green, oolong and black teas of C.
sinensis screened for AOP are listed in Table 1. Tropical herbal teas
of misai kucing (O. aristatus), lemon grass (C. citratus), guava (P.
guajava), bitter gourd (Momordica charantia), lemon myrtle
(Backhousia citriodora), mas cotek (Ficus deltoidea), pegaga (Centella
asiatica) and rooibos (A. linearis); temperate herbal teas of
rosemary (Rosmarinus ofcinalis), peppermint (Mentha piperita),
mint (Mentha spicata), chamomile (Matricaria recutita) and oregano
(Origanum vulgare); and green, oolong and black teas of C. sinensis
were purchased from the supermarket. Teas of getto (Alpinia
zerumbet) and rang jeud (T. laurifolia) were obtained fromOkinawa
(Japan) and Bangkok (Thailand), respectively. Teas of legundi (Vitex
negundo) and asam gelugor (Garcinia atroviridis) were obtained
fromForest Research Institute Malaysia (FRIM). Dried owers of jin
Journal of Food Composition and Analysis 23 (2010) 185189
A R T I C L E I N F O
Article history:
Received 23 October 2008
Received in revised form 11 September 2009
Accepted 20 October 2009
Keywords:
Tropical teas
Temperate teas
Herbal teas
Camellia sinensis
Antioxidant activity
Food analysis
Food composition
A B S T R A C T
Antioxidant properties (AOP) of thirteen tropical and ve temperate herbal teas were screened.
Comparisons were made with green, oolong and black teas of Camellia sinensis. The AOP studied were
total phenolic content, radical-scavenging activity, ferric-reducing power and ferrous ion-chelating (FIC)
ability. Tropical herbal teas were more diverse in types and more variable in AOP values than temperate
herbal teas. Herbal teas generally had lower antioxidant values than teas of C. sinensis. Exceptions were
lemon myrtle, guava and oregano teas with AOP comparable to black teas. FIC ability of mint and
peppermint teas was signicantly stronger than all C. sinensis teas.
2010 Elsevier Inc. All rights reserved.
* Corresponding author. Tel.: +60 3 55146103; fax: +60 3 55146364.
E-mail address: Lim.Yau.Yan@sci.monash.edu.my (Y.Y. Lim).
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doi:10.1016/j.jfca.2009.10.002
yin hua (Lonicera japonica) were purchased from the Chinese
medicine shop. In this study, tropical herbal teas included several
plant species from the subtropics such as B. citriodora, A. linearis, A.
zerumbet and L. japonica. Teas of C. sinensis, a tropical and sub-
tropical species, are not considered as herbal teas.
2.2. Extraction of teas
In tea extraction, 1 g of tea in powder form was extracted with
50 mL boiling water. Infusions were allowed to steep for 1 h with
continuous swirling. Extracts were ltered and stored at 4 8C for
further analysis. Analyses of aqueous tea extracts were done in
triplicate.
2.3. Extraction efciency
Extraction efciency of boiling water was tested with Boh green
tea. After the rst extraction, the residues were ltered and
extracted successively for the second and third time. Extraction
efciency in percent was based on total phenolic content of rst,
second and third extractions.
2.4. Antioxidant properties of teas
Total phenolic content (TPC) was determined using the Folin-
Ciocalteu assay (Ka hko nen et al., 1999). Samples (300 mL, in
triplicate) were introduced into test tubes followed by 1.5 mL of
Folin-Ciocalteus reagent (10 times dilution) and 1.2 mL of sodium
carbonate (7.5%, w/v). The tubes were allowed to stand for 30 min
before absorbance at 765 nm was measured. TPC was expressed as
gallic acid equivalent (GAE) in mg per 100 g material. The
calibration equation for gallic acid was y = 0.0111x 0.0148
(R
2
= 0.9998), where y is the absorbance and x is the gallic acid
concentration in mg/L.
Radical-scavenging activity (RSA) was assessed using the 2,2-
diphenyl-1-picrylhydrazyl (DPPH) assay (Miliauskas et al., 2004).
Different dilutions of extracts (1 mL) were added to 2 mL of DPPH
(5.9 mg per 100 mL methanol). After 30 min, absorbance was
measured at 517 nm. RSA, expressed as ascorbic acid equivalent
antioxidant capacity (AEAC) in mg ascorbic acid/100 g, was
calculated as IC
50(ascorbate)
/IC
50(sample)
10
5
. IC
50
of ascorbic acid
was 0.00387 mg/mL.
For assessing ferric-reducing power (FRP), the assay described
by Chu et al. (2000) was adapted. Different dilutions of extracts
(1 mL) were added to 2.5 mL phosphate buffer (0.2 M, pH 6.6) and
2.5 mL of potassium ferricyanide (1%, w/v). The mixture was
incubated at 50 8C for 20 min. After trichloroacetic acid solution
(2.5 mL, 10%, w/v) was added, the mixture was separated into
aliquots of 2.5 mL and diluted with 2.5 mL of water. To each diluted
aliquot, 500 mL of ferric chloride solution (0.1%, w/v) was added.
After 30 min, absorbance was measured at 700 nm. FRP of extracts
was expressed as mg GAE/g. The calibration equation for gallic acid
was y = 16.767x (R
2
= 0.9974), where y is the absorbance and x is
the gallic acid concentration in mg/mL.
The ferrous-ion-chelating (FIC) assay was adapted from Singh
and Rajini (2004). FIC ability was determined by mixing FeSO
4
(0.1 mM, 1 mL) with different dilutions of extracts (1 mL), followed
by ferrozine (0.25 mM, 1 mL). Absorbance was measured at 562 nm
after 10 min. The ability of extracts to chelate ferrous ions was
calculated as chelating effect % = (1 A
sample
/A
control
) 100. FIC
ability was expressed as chelating EC
50
(CEC
50
) in mg/mL, or the
effective concentration of extract to chelate ferrous ions by 50%.
3. Results and discussion
Based on three successive extractions of Boh green tea (1 g)
with boiling water (50 mL) for 1 h, TPC of rst, second and third
extractions was found to be 75 1.3, 17 0.5 and 8 0.7%,
respectively. As the yield of rst extraction was high, all analyses
of teas were extracted with boiling water. Signicantly higher yields
of hot water than cold water extraction of green tea (Lin et al., 2008)
and stronger RSA of oolong tea extracted with hot water of increasing
temperature (Su et al., 2006) have been reported. For green, oolong
and black teas, extraction with water at 100 8C for 3 min yielded
Table 1
Types of tropical and temperate herbal teas, and green, oolong, and black teas of Camellia sinensis screened for antioxidant properties.
Type of tea Species (family) Part used and life-form Company, country
Tropical herbal tea
Lemon myrtle Backhousia citriodora (Myrtaceae) Leaf, shrub Boh, Malaysia
Guava Psidium guajava (Myrtaceae) Leaf, tree Organic Herb, Malaysia
Legundi Vitex negundo (Lamiaceae) Leaf, tree FRIM, Malaysia
Rooibos Aspalathus linearis (Leguminosae) Leaf, shrub Dr Nortier, S. Africa
Jin yin hua Lonicera japonica (Caprifoliaceae) Flower, vine Ren ZK, Malaysia
Misai kucing Orthosiphon aristatus (Lamiaceae) Leaf, herb Pure Herb, Malaysia
Pegaga Centella asiatica (Apiaceae) Leaf, herb BZest, Malaysia
Mas cotek Ficus deltoidea (Moraceae) Leaf, shrub CBintik, Malaysia
Bitter gourd Momordica charantia (Cucurbitaceae) Fruit, vine Everjoy, Vietnam
Getto Alpinia zerumbet (Zingiberaceae) Leaf, herb Nakazen, Japan
Asam gelugor Garcinia atroviridis (Guttiferae) Leaf, tree FRIM, Malaysia
Rang jeud Thunbergia laurifolia (Thunbergiaceae) Leaf, vine TriSiam, Thailand
Lemon grass Cymbopogon citratus (Graminae) Stem, herb Organic Herb, Malaysia
Temperate herbal tea
Oregano Origanum vulgare (Lamiaceae) Leaf, herb McCormack, US
Mint Mentha spicata (Lamiaceae) Leaf, herb McCormack, US
Peppermint Mentha piperita (Lamiaceae) Leaf, herb Twinings, UK
Rosemary Rosmarinus ofcinalis (Lamiaceae) Leaf, herb McCormack, US
Chamomile Matricaria recutita (Asteraceae) Flower, herb Dilmah, S. Lanka
Camellia tea
Green Tea (g) Camellia sinensis (Theaceae) Leaf, shrub Boh, Malaysia
Green Tea (g) Sea Dyke, China
Ti Kuan Yin (o) Sea Dyke, China
Yellow Label (b) Lipton, Thailand
Cha Wang (o) An Xi, China
Bukit Cheeding (b) Boh, Malaysia
Cameron Highlands (b) Boh, Malaysia
E.W.C. Chan et al. / Journal of Food Composition and Analysis 23 (2010) 185189 186
higher total avan-3-ol content than extraction with water at 60 and
80 8C (Horzic et al., 2009).
Tropical herbal teas screened were diverse in types, parts used
and morphology. They comprised leaves, owers, fruits and stems
of 13 species of trees, shrubs, vines and herbs belonging to 13
genera and 11 families (Table 1). AOP were variable with TPC,
AEACand FRP values ranging from644 to 7560 mg GAE/100 g, 354
to 13,600 mg AA/100 g and 3 to 61 mg GAE/g, respectively (Table
2). Teas of Myrtaceae (lemonmyrtle and guava) displayed the best
TPC, RSAand FRP, with values ranging from5930 to 7560 mg GAE/
100 g, 7430 to 13,600 mg AA/100 g and 35 to 61 mg GAE/g,
respectively. Getto, mas cotek, misai kucing, lemon myrtle and
guava teas displayed strong FIC ability with CEC
50
values ranging
from 0.9 to 1.2 mg/mL. Their strong AOP could be due to the
content and composition of major phenolic compounds (Table 3).
Temperate herbal teas screened were less diverse in types, parts
used and morphology. They comprised leaves and owers of ve
species of herbs belonging to four genera and two families,
Lamiaceae being dominant (Table 1). AOP were less variable with
TPC, AEAC and FRP values ranging from 1370 to 5860 mg GAE/
100 g, 966 to 7240 mg AA/100 g and 8 to 49 mg GAE/g, respectively
(Table 2). Temperate herbal teas of oregano, mint and peppermint
Table 2
Total phenolic content (TPC), ascorbic acid equivalent antioxidant capacity (AEAC), ferric-reducing power (FRP), and chelating EC
50
(CEC
50
) of tropical and temperate herbal
teas.
Type of tea TPC (mg GAE/100g) AEAC (mg AA/100g) FRP (mg GAE/g) CEC
50
(mg/mL)
Tropical herbal tea
Lemon myrtle 7560126a 13 600792a 610.6a 1.20.2a
Guava 5930584b 7430752b 352.7b 1.20.2a
Legundi 5420404b 4900549c 302.0c 1.90.1b
Rooibos 3750235c 3020456e 200.4e 4.71.2d
Jin yin hua 3630169c 3850224d 210.5d 2.00.2b
Misai kucing 278071d 4160362cd 220.9d 1.10.5a
Pegaga 2040214e 1340613f 121.2f 2.70.1c
Mas cotek 202087e 108066f 5.90.3h 1.00.0a
Bitter gourd 143030f 110063f 100.4g 2.00.1b
Getto 85421g 53656g 3.50.3i 0.90.1a
Asam gelugor 80694g 26215i 2.90.2j >7.0e
Rang jeud 80550g 59129g 4.30.5i 4.00.5d
Lemon grass 64420h 35431h 3.10.2j 4.70.3d
Temperate herbal tea
Oregano 5860379a 7240309a 491.6a 0.80.2b
Mint 435020b 4430310c 310.4c 0.30.0a
Peppermint 4210139b 6040311b 370.3b 0.40.1a
Rosemary 281089c 309097d 201.2d 2.10.5c
Chamomile 137037d 96652e 8.10.2e 3.30.1d
Camellia tea
Green Tea (g) 14,1201810a 25 0002780a 14311a 1.80.3c
Green Tea (g) 11,3701480b 18 4601740b 8411b 1.40.1b
Ti Kuan Yin (o) 9090457c 16 1702480b 694.2c 1.90.2c
Yellow Label (b) 8490803c 11 5501150cd 533.0e 1.70.1c
Cha Wang (o) 7500462d 14 4503046bc 592.3d 1.80.5c
Bukit Cheeding (b) 7410120d 10 300563d 533.0e 1.00.2a
Cameron Highlands (b) 6060543e 75101260e 362.4f 1.50.2bc
Teas are ranked based on TPC and values are means SD (n= 3). For each column, values followed by the same letter are not statistically different at P <0.05 as measured by the
Tukey HSDtest. ANOVA does not applying between tropical, temperate, and Camellia teas. Green (g), oolong (o), and black (b) teas of Camellia sinensis were used as positive controls.
Table 3
Major phenolic compounds of herbal teas studied.
Tea Major phenolic compound Source
Tropical
Guava GA and quercetin Liang et al. (2005)
Legundi Orientin and luteolin Tiwari and Tripathi (2007)
Rooibos Aspalathin, rutin, and orientin Bramati et al. (2002)
Misai kucing RA Akowuah et al. (2005)
Pegaga Asiatic acid and asiaticoside Kim et al. (2009)
Bitter gourd GA and caffeic acid Kubola and Siriamornpun (2008)
Lemon grass Chlorogenic acid Marques and Farah (2009)
Temperate
Oregano RA and hispidulin Zheng and Wang (2001)
Mint Eriocitrin, luteolin, and RA Damien Dorman et al. (2003)
Peppermint Eriocitrin and RA Areias et al. (2001)
Rosemary Carnosic acid, rosmanol, and RA Zheng and Wang (2001)
Chamomile Apigenin and quercetin McKay and Blumberg (2006)
Camellia
Green tea EGCG and EGC Horzic et al. (2009)
Oolong tea EGCG, EGC, and GA Horzic et al. (2009)
Black tea GC, EGCG, and GA Horzic et al. (2009)
Abbreviations: GA=gallic acid, RA=rosmarinic acid, EGCG=epigallocatechin gallate, EGC=epigallocatechin, and GC=gallocatechin.
E.W.C. Chan et al. / Journal of Food Composition and Analysis 23 (2010) 185189 187
displayed strong AOP with outstanding CEC
50
values of 0.8, 0.3 and
0.4 mg/mL, respectively. With the exception of chamomile tea
(Asteraceae), rosmarinic acid is one of the major phenolic
compounds in all the other herbal teas (Lamiaceae) (Table 3).
The strong AOP of temperate herbal teas might be attributed to the
predominance of rosmarinic acid and other phenolic acids.
With higher percentage of teas having strong AOP, particularly
in FIC ability, temperate herbal teas investigated were superior to
tropical herbal teas. Temperate herbal teas with strong primary
AOP (RSAand FRP) also had strong secondary AOP (FIC ability). This
was not evident in tropical herbal teas as some teas with strong
primary AOP displayed poor secondary AOP (e.g. rooibos tea) and
vice versa (e.g. getto tea). RSA and FRP are measures of the
hydrogen and electron-donating abilities of primary antioxidants,
respectively (Lim et al., 2007). FIC ability measures the ability of
secondary antioxidants to chelate metal ions. Primary antioxidants
prevent oxidative damage by directly scavenging free radicals,
while secondary antioxidants act indirectly by preventing the
formation of free radicals through Fentons reaction.
Teas of C. sinensis had TPC, AEAC, FRP and CEC
50
values ranging
from 6060 to 14,120 mg GAE/100 g, 7510 to 25,000 mg AA/100 g,
36 to 143 mg GAE/g and 1.0 to 1.9 mg/mL, respectively (Table 2).
Ranking of TPC, RSA, FRP and FIC ability was green > oolong -
black; green oolong > black; green > oolong > black; and
black > green oolong, respectively. Findings from this study
deviated from Yen and Chen (1995) who reported that ranking of
RSA was green > oolong > black, and ranking of reducing power
was oolong > green > black; and fromvonGadowet al. (1997) and
Yokozawa et al. (1998), who reported that ranking of RSA was
green > black > oolong. Green teas have been reported to have
signicantly higher TPC, RSA and FRP, but poorer FIC ability, than
black teas (Chan and Lim, 2006; Chan et al., 2007).
The outstanding AOP of green, oolong and black teas of C.
sinensis could be attributed to their high avanol content of
epigallocatechin gallate (EGCG), epigallocatechin (EGC) and
gallocatechin (GC) (Table 3). Recently, Horzic et al. (2009)
reported that green, oolong and black teas, extracted with
deionised water heated to 100 8C for 3 min, had total identied
avan-3-ol contents of 999, 666 and 672 mg/L, respectively.
Temperate herbal teas of linden and chamomile yielded avan-3-
ol contents of only 127 and 22 mg/L, respectively.
Compared to teas of C. sinensis, tropical and temperate herbal
teas generally had lower antioxidant values. Exceptions were
lemon myrtle, guava and oregano teas with AOP comparable to
black teas. CEC
50
values of mint and peppermint were signicantly
larger than all C. sinensis teas. FIC ability of oregano, getto, mas
cotek, misai kucing, lemon myrtle and guava teas was comparable
to black teas. It can be seen that some tropical and temperate
herbal teas had AOP that are superior or comparable to green,
oolong and black teas.
In conclusion, tropical herbal teas were more diverse in types
and more variable in AOP than temperate herbal teas. Herbal teas
generally had lower antioxidant values than teas of C. sinensis.
Acknowledgements
The authors are thankful to Monash University Sunway Campus
(MUSC) Malaysia for nancial support of this project, to Forest
Research Institute Malaysia (FRIM) for providing tea samples of
legundi and asam gelugor, and to Dr. Mami Kainuma for
purchasing the getto tea from Okinawa, Japan.
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