You are on page 1of 6

Food Chemistry 110 (2008) 859–864

Contents lists available at ScienceDirect

Food Chemistry
journal homepage:

In vitro antimicrobial activities against cariogenic streptococci and their antioxidant capacities: A comparative study of green tea versus different herbs
Tzung-Hsun Tsai a, Tsung-Hsien Tsai b, You-Chia Chien a, Chi-Wei Lee c, Po-Jung Tsai d,*

Department of Dentistry, Keelung Chang-Gung Memorial Hospital, Keelung, Taiwan Department of Dermatology, Taipei Medical University – Wanfang Hospital, Taipei, Taiwan c Institute of Biotechnology, Yuanpei University, Hsinchu, Taiwan d Department of Human Development and Family Studies, National Taiwan Normal University, No. 162, Sec. 1, Hoping E. Road, Taipei 10610, Taiwan

a r t i c l e

i n f o

a b s t r a c t
The antimicrobial activity against cariogenic bacteria, total antioxidant capacity and phenolic constituents of methanolic extracts from 11 herbs were investigated and compared with those of green tea (Camellia sinensis). Among the 12 tested herbs, eight herbal extracts could inhibit the growth of Streptococcus sanguinis. Jasmine, jiaogulan, and lemongrass were the most potent, with minimum inhibitory concentrations (MIC) of 1 mg/ml, while green tea was less effective, with a MIC of 4 mg/ml. Among them, only rosemary could inhibit the growth of S. mutans at a MIC of 4 mg/ml. Total antioxidant capacities of herbal extracts were analyzed by three different assays, including 2,2-diphenyl-1-picrylhydrazyl (DPPHÁ) radical scavenging activity, trolox equivalent antioxidant capacity (TEAC) and oxygen radical absorbance capacity (ORAC). Regardless of the assays used, green tea exhibited the highest antioxidant capacity, followed by osmanthus. Wide variations in total phenolics and total flavonoids of herbal tea extracts were observed. Chlorogenic acid was detected in high amount in honeysuckle and duzhong. These data suggest that rosemary is a potent inhibitor of oral streptococci, and green tea and osmanthus may be effective potential sources of natural antioxidants. Ó 2008 Elsevier Ltd. All rights reserved.

Article history: Received 22 September 2007 Received in revised form 7 January 2008 Accepted 25 February 2008

Keywords: Herbal teas Cariogenic streptococci Antioxidant capacity

1. Introduction Green tea (Camellia sinensis) has been considered, in traditional Chinese medicine, as a healthful beverage, and PET-bottled green tea has recently become popular with the Taiwanese. Recent human studies suggest that green tea may contribute to a reduction in the risk of cardiovascular disease and some forms of cancer, as well as to the promotion of oral health and other physiological functions, such as antioxidant and antibacterial activities, antihypertensive effect and body weight control (Cabrera, Artacho, & Gimenez, 2006). In Taiwan, green tea leaves are reprocessed and scented with osmanthus (Osmanthus fragrans), rose (Rosa damascena) or jasmine (Jasminum sambac) flowers, combining fragrance and sweet taste. Herbs have been used for many purposes, including medicine, nutrition, flavouring, beverages, and fragrance. Juhua (Chrysanthemum morifolium) and honeysuckle (Lonicera japonica) are used as Chinese natural medicine and possess antibacterial and anti-inflammatory activities (Miyazawa & Hisama, 2003; Park et al., 2005). Duzhong (Eucommia ulmoides) and jiaogulan (Gynostemma pentaphyllum) teas are commonly consumed and regarded as functional health foods in Taiwan. Duzhong exhibits antioxidant
* Corresponding author. Tel.: +886 2 23636425x55; fax: +886 2 23639635. E-mail address: (P.-J. Tsai). 0308-8146/$ - see front matter Ó 2008 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodchem.2008.02.085

activity in various lipid peroxidation models (Yen & Hsieh, 2000) and anti-diabetic potential (Lee et al., 2005). Jiaogulan has a variety of biological activities, including anti-inflammatory, immunopotentiating, and antioxidant effects (Norberg et al., 2004). In addition to the popular scented teas and traditional herbal teas in Taiwan, various herbal teas have become increasingly popular because of their fragrance and antioxidant activities. For example, lavender (Lavandula angustifolia) is a well-known aromatic herb and exhibits antibacterial and anti-inflammatory properties (Hajhashmi, Ghannadi, & Sharif, 2003; Rota, Carraminana, Burillo, & Herrera, 2004). Lemongrass (Cymbopogon citratus) has been found to possess antioxidant activity and is recommended to treat digestive disorders, diabetes, and inflammation (Cheel, Theoduloz, Rodrigues, & Schmeda-Hirschmann, 2005). Mate (mä_ta  ; Ilex paraguariensis) is a popular beverage of many South American countries; extracts contain polyphenols and flavonoids and exhibit strong antioxidant activity (Gugliucci, 1996). Dental caries, a common type of dental disease, is associated with microorganisms present on the tooth surface in dental plaque. Mutans streptococci, including Streptococcus mutans, S. sobrinus, S. cricetus, S. rattus, and S. ferus, participate in the formation of dental biofilm and play a significant role in the initiation of dental caries. S. mutans and S. sobrinus are regularly isolated from humans (Hardie, 1998). S. sanguinis (formerly S. sanguis) is colonized on the

the polyphenols and catechins constitute the most interesting group of tea leaf components: (À)-epigallocatechin gallate. Numerous studies have demonstrated that spices and herbs possess antibacterial properties against food-borne bacteria and fungi (Roy & Lai. & Hamada. All other chemicals were of analytical-grade purity. (À)-gallocatechin. Fujiwara. mutans (BCRC10793) were obtained from the Bioresource Collection and Research Center. based on electron transfer reaction. Rosalen. 2005. For example. cocoa bean husk extract (Ooshima et al. one of the objectives of this study was to determine three major flavonols (kaempferol.4. tocopherols. jiaogulan. quercetin. jasmine. Curin & Andriantsitohaina. mutans with water-soluble glucan-synthesizing S. These bacteria were cultured in an anaerobic atmosphere. 2005). Louis.-H. mate. including juhua. 2004). Tanabe. 0. After incubation for 48 h at 37 °C under anaerobic condition.7 8. USA).0 24. duzhong. In this study. jiaogulan.0 16.3 21. 2000). rutin. Taiwan). lavender. Determination of anti-streptococcal activity The methanolic extracts were tested against oral pathogens by determining minimum inhibitory concentration (MIC) values obtained by a microdilution broth method. Jasminum sambac Chrysanthemum morifolium Lavandula angustifolia Osmanthus fragrans Rosa damascena Eucommia ulmoides Oliv.5% yeast extract. We recently reported that aqueous and methanolic extracts of rosemary (Rosmarinus officinalis) inhibited S. MO.05 absorbance. 2000). caffeic acid. Triplicate samples were performed for each test concentration. C. (À)-epigallocatechin. Folin-Ciocalteu phenol reagent was added to the reconstituted samples and held for 3 min. Materials Streptococcus sanguinis (BCRC15273) and S.. w/w) 20. Liao & Liu. Herbs and herbal extracts contain different phytochemicals with biological properties that promote human health and help reduce the risk of chronic disease (Craig. Larson. and rosemary) commonly consumed in Taiwan.4 2. and to compare them with green tea for possible use as anticariogenic agents and natural antioxidants.1. Di. & Bowen. The experiments were performed in duplicate. and myricetin) and two phenolic acids (chlorogenic acid and caffeic acid) in various herbs commonly consumed in Taiwan. especially food extracts. Ou. sanguinis yields firm adhesion (Tamesada. 2007).5 8. osmanthus. Detroit. sanguinis and S. rose. Tsai. Preparation of extracts Twelve dried herbs. sinensis is a dietary source of antioxidant nutrients. 2. 2004).6 13.).05% L-cysteine. Kanda. mate. the growth of streptococci was estimated spectrophotometrically at 610 nm using a microtitre plate reader. (St. The aim of this study was to characterize the antimicrobial activities against oral pathogens. for example in the generation of different radicals and target molecules. / Food Chemistry 110 (2008) 859–864 tooth surface during the early stage of dental plaque formation. rose.06–8 mg/ml of methanolic extracts. ascorbic acid. and oxygen radical absorbance capacity (ORAC) assay. a parallel series of mixtures with uninoculated broth was prepared. were purchased from a local supermarket in Taipei City (Taipei. tems (Becton Dickinson Microbiology Systems. and selected phenolic components of herbs (juhua. Total phenolic compound analysis The amount of total phenolics in herbal extracts was evaluated using spectrophotometric analysis with Folin-Ciocalteu reagent (Zheng & Wang.2-diphenyl-1-picrylhydrazyl (DPPHÁ) radical-scavenging analysis and trolox equivalent antioxidant capacity (TEAC) assay. In sterile 96-well microtiter plates. To adjust for the interference by plant pigments.4 10. Briefly. 2. honeysuckle. 100 ll of herbal extract were diluted with broth and placed into the well containing 100 ll of bacterial suspension in broth.7 16.4 26. three analyses were performed to determine the total antioxidant activities of herbs and green tea. sobrinus growth and its glucosyltransferase activity (Tsai.7 26. Even though many herbs are known to be sources of phenolic compounds. green tea. Tsai et al. Chlorogenic acid. honeysuckle. Kawabata. mutans were. lemongrass. and rosemary. Among these. quercetin. USA) containing 0. Li. 2001). cultured in Brain Heart Infusion (BHI) broth and Tryptic Soy Broth (TSB) (Difco Laboratories. respectively.4 26. After extraction. using BBL GasPak sys- . Camellia sinensis Gynostemma pentaphyllum Cymbopogon citratus Ilex paraguariensis Rosmarinus officinalis Part examined Flowers Flowers Flowers Flowers Flowers Flowers Leaves Leaves Leaves Leaves Leaves Leaves Extract yield (%. their compositional data are still insufficient. mutans from overnight cultures were adjusted to 1 Â 106 colony-forming units (CFU)/ml. Cockeysville. The extraction yield of methanolic extract was presented as % weight (gram of methanolic extract for each 100 g of dry herbs) and the results are shown in Table 1. and apple polyphenols (Yanagida. (À)epicatechin. MI. Cury. Md. and myricetin were purchased from Sigma Chemical Co. kapempferol. jasmine. Taiwan). but there are few data or observations concerning the antimicrobial activity of herbal teas against oral pathogens. known as bioactive agents. Several methods have been developed to measure the total antioxidant capacities of food and beverages. and non-nutrient phytochemicals generally classified as flavonoids. Food Industry Research and Development Institute (Hsinchu. Phenolic acids and flavonoids. S.2. Tsai. including the 2. antioxidant capacities. Coculture of transformed S. have been shown to have potentially useful anticariogenic properties. and 1% sucrose. Matsudaira. & Ho. Briefly.3. based on hydrogen atom transfer reaction (Huang. Hence. Twofold serial dilutions were made in broth over a range to give the final concentrations of 0. 2001. propolis (Koo. duzhong. lemongrass. frequently occur in herbal plants (Miean & Mohamed. Materials and methods 2. The combined methanolic solution was centrifuged at 12. 2007). The MIC was defined as the minimum concentration of test compound limiting turbidity to <0. lavender. as previously described (Tsai. on resident oral bacteria. S. & Ho. & Prior. osmanthus. 1992). Park. these assays differ in their chemistry. sanguis and S. The methanolic extract was reconstituted in dimethyl sulfoxide (DMSO) to a concentration of 400 mg/ml for subsequent experimentation. 1988). 2005).000g for 10 min and evaporated on a rotary evaporator. the mixture was filtered and the residue was re-extracted with 50 ml of fresh methanol overnight. and (À)-catechins (Graham. such as carotenoids. Wen. 2. & Oliveria Cordeiro. 1999. (À)-epicatechin gallate. 2002). There is increasing interest in the effect of natural compounds.860 T. The dried herbs (10 g) were extracted with 50 ml of methanol at room temperature for 3 h. Then 2 ml of 10% (w/v) sodium carbonate solution were added and Table 1 Characteristics of various herbal teas Common name Honeysuckle Jasmine Juhua Lavender Osmanthus Rose Duzhong Green tea Jiaogulan Lemongrass Mate Rosemary Botanical name Lonicera japonica Thunb.

Moon & Lee.6.-H. Detection was at 325 nm for caffeoyl derivatives. Giberti. MA. 2. Tsai. 150 ll of fluorescein (30 . IL. & Ho. 2.5 ml of 1 M NaOH were sequentially added at 0. were identified by using a reverse-phase high-performance liquid chromatography (HPLC) method (Filip. and at 263 nm for kaempferol.05.60 -dihydroxyspiro [isobenzofuran-1[3H]. 3. The reaction mixture was mixed and left to stand for 10 min. sanguinis.2-azobis(2-amidinopropane) dihydrochloride (AAPH) solution was added as a peroxyl radical generator. 10 min. 2003).7. 90 [9H]-xanthen]-3-one) solution were put into each well and the plate was incubated at 37 °C for 30 min in the dark.0 statistical package (Chicago. Lopez. methanol/ acetic acid 98:2 (v/v)): 15% B to 40% B. Scavenging of diphenyl-picrylhydrazyl (DPPHÁ) radicals The 2. The antioxidant activity of the sample was calculated as follows: % DPPHÁ radical-scavenging activity = (1À[Asample À Ablank of sample/ADMSO À Ablank of DMSO]) Â 100. mutans and S. Wals.. The absorbance at 540 nm was determined against a blank of ethanol.20 azinobis (3-ethylbenzothiazoline sulfonate (ABTS+) according to the manufacturer’s instruction (Randox Laboratories Ltd. and Prior (2001). / Food Chemistry 110 (2008) 859–864 861 allowed to stand at room temperature for 30 min. 250 Â 4. Coussio. Finally. The flavonoid content in each extract was then calculated by a standard curve prepared with catechin and expressed as milligrams of catechin equivalents (CE) per gram of solid of extract. caffeoyl derivatives (chlorogenic acid and caffeic acid) and flavonoids (rutin.5.10. 5 and 6 min. Waltham. myricetin. at 254 nm for rutin.075 ml of 10% AlCl3. 0. and 75% B to 85% B.9. Lagerhausstr. Trolox was used as a control antioxidant standard. 25 ll of the 2. Yu. 5 min. Pearson correlation analysis was used to evaluate the relationships among the variables of interest. Oxygen radical absorbing capacity (ORAC) assay The ORAC assay was performed as described by Ou. Data were subjected to analysis of variance. mutans. The flow rate was 1 ml/min and the injection volume was 20 ll. Results and discussion 3. 2007). The phenolic compounds were expressed as mg/g of methanolic extract. only rosemary could inhibit the growth of S. 2001). Chun. sanguinis 4 1 >8 >8 >8 >8 4 4 1 1 4 2 S. 1. The absorbance at 510 nm was measured. The absorbance at 765 nm was measured. & Ho. Antioxidants in the sample suppress the development of colour to an extent that is proportional to their concentration. ABTS+ has a stable blue-green colour. As shown in Table 2. kapempferol. The final ORAC values were calculated using the differences of area under the quenching curves of fluorescein between a blank and a sample. Then.8. Among the 12 tested herbs. A P-value of less than 0.0) were pipetted into a 96-well flat-bottomed plate. 2. Briefly.175 ml of dd H2O was added to the above reacting mixture and thoroughly mixed. Following this. water/acetic acid 98:2 (v/v) and solvent B. 2. Statistical analysis Data were presented as means + SD. sobrinus >8 >8 >8 >8 >8 >8 >8 >8 >8 >8 >8 4* *Data for S. consisting of binary pump and UV photodiode array detector and equipped with a ODS Hypersil-C18 (5 mm. and quercetin. The HPLC analysis was performed using a Hewlett-Packard HPLC system (HP 1100 series. 25 ll of sample dissolved in 100 ll of 75 mM phosphate buffer (pH 7. Identification of individual phenolics was based on the comparison of the retention time and UV spectrum of unknown peaks with those of reference authentic standards.25 ml of optimally diluted sample was put into the tube containing 1 ml of double-distilled water (dd H2O). Briefly.05 was considered statistically significant. Tsai et al. and myricetin). .T. Briefly. namely. Crumlin. and the fluorescence readings were taken immediately by a Zenyth 3100 microplate multimode detector (Anthos Labtec Instruments Inc. Waldbronn.6 mm) column (Thermo Electron Corporation. The unit was expressed as millimoles of trolox equivalents (TE) per gram of methanolic herbal extracts. The total phenolic content was calculated by a standard curve prepared with gallic acid and expressed as milligrams of gallic acid equivalents (GAE) per gram of solid of extract. Germany).2-diphenyl-1-picrylhydrazyl (DPPHÁ) radical-scavenging capacity of each herbal methanolic extract was measured as described earlier (Tsai. & Ferraro. We previously demonstrated that aqueous and methanolic extracts of rosemary exhibit antimicrobial activity against oral bac- Table 2 The minimal inhibition concentration (MIC) of methanolic extracts of various herbs against oral pathogens Herbs MIC (mg/ml) S. the crude methanolic extracts from herbs and green tea leaves were evaluated for growth inhibitory activity against S. and 0. mutans Honeysuckle Jasmine Juhua Lavender Osmanthus Rose Duzhong Green tea Jiaogulan Lemongrass Mate Rosemary >8 >8 >8 >8 >8 >8 >8 >8 >8 >8 >8 4 S. 20 ll of each sample or 100% DMSO (as a negative control) were allowed to react with 200 ll of freshly prepared 200 lM DPPHÁ ethanolic solution in a 96-well microplate. quercetin. Kim. which is measured at 600 nm. Trolox equivalent antioxidant capacity (TEAC) assay The radical-scavenging capacity of the methanolic extracts of selected herbs and green tea leaves was measured against 2. 0. 2007).75 ml of 5% NaNO2.1. 30 min. Tsai. 0. USA). sobrinus were reported previously (Tsai. 2. 40% B to 75% B. 2. The HPLC method used the following gradient elution programme: (solvent A. Austria) every 5 min for 120 min. Determination of total flavonoids Total flavonoids were measured according to a colorimetric assay (Kim. Then. Anti-streptococcal activity The elimination of cariogenic bacteria from the oral cavity using antibacterial agents is one of the primary strategies for the prevention of dental caries. Analysis of selected phenolic compounds in herbs The selected phenolics in methanolic herbal extracts. Statistical analysis was conducted using the SPSS 12. UK) This assay was based on ABTS+ incubated with metmyoglobin and hydrogen peroxide to generate the radical cation ABTS+. Results were expressed in millimoles of trolox equivalents (TE) per gram of solid of herbal tea extract. HampschWoodill. USA). and means were separated using Fisher’s least significant difference (LSD) test at P = 0.

sinensis).g.934.. As with total phenolics in herbal teas tested. but no appreciable activity against S. sobrinus. in decreasing order. methanolic extract from green tea possessed antimicrobial activity against S.4-fold higher than in lavender. Numerous studies have demonstrated that green tea.. The total antioxidant capacities of the herbal methanolic extracts and the ranking order for each assay are shown in Table 3. The high level of polyphenolics and antioxidant activity in green tea has been 160 A LSD0. 1998). lemongrass (Cheel et al. green tea. and Himejima (1992) demonstrated that volatile flavour compounds in green tea had inhibitory activity against S.001) and the DPPHÁ-scavenging capacity and TEAC values (r = 0. p < 0. whereas the TEAC values and ORAC values (r = 0. 2000) exhibited antioxidant properties. and duzhong (Yen & Hsieh. When the ORAC assay was used to provide a ranking order of antioxidant activity. 2004). The total flavonoid content of sweet osmanthus was $9. from these preparations. 1992. in decreasing order. mutans. mate (Bixby.3.. sanguinis at MIC values of 4 and 2 mg/ml.05 = 1. The herbal extracts exhibited different antioxidant capacities in relation to the method applied. for the remaining herbs analyzed was: rose > mate > rosemary > honeysuckle > juhua > duzhong > lemongrass > jasmine > jiaogulan > lavender. and oligomeric catechins. the presence of sucrose in culture broth. Honeysuckle. and procedures for extraction. p < 0. Furthermore. the DPPH radical.97 Total phenolics (mg GAE/g extract) 140 120 100 80 60 40 20 0 Total flavonoids (mg CE/g extract) 250 B LSD0. Menini. The data obtained using the DPPHÁ-scavenging capacity and ORAC values had the best correlation coefficients (r = 0. serotypes of S.734.32 mg/ml of methanolic extract. & Gugliucci.. Tsai. sobrinus (Hamilton-Miller. rose. 1. showing the wide variance of total phenolic concentrations in tested herbal teas. Further studies are needed to explore the potential values of rosemary or its components as natural antiplaque or antigingivitis agents. 2005). depending on the assay. Notably. and lemongrass demonstrated preferential antimicrobial activity against S. The yield of methanolic extract from green tea leaves was 21. Our results showed that methanolic extract from green tea did not completely inhibit the growth of S. the levels of total flavonoids among the herbal teas resulted in significant differences. Kubo.7 and 225 mg CE/g of extract. thus the same material ranked differently.6% (w/ w) (Table 1). Rasheed & Haider.-H. reported (Cabrera et al. However.5 times higher than that of jasmine. Green tea had the greatest antioxidant capacity of all the herbs used in each assay. Sweet osmanthus had the highest total flavonoid content. 1993). whereas jasmine had the lowest. The level of total phenolics in green teas was $4. the TEAC values in decreasing order for the remaining herbs analyzed were mate > rosemary > rose > honeysuckle > jiaogulan > duzhong > juhua > lemongrass > lavender > jasmine. there was no significant difference in DPPHÁ-scavenging capacity between green tea and osmanthus. 3. 2007). 2006). mutans and S. sources and manufacturing methods of tea. Total flavonoid levels ranged between 23. Theoretically.862 T.scavenging capacity. Several studies also showed that juhua (Kim & Lee. jiaogulan (Norberg et al.05= 2. However. lavender. 1996). 1. sanguinis with MIC values of 4 mg/ml. mutans (Kubo et al. & Ho. Tsai et al. Green tea was found to have the highest total phenolics among the 12 herbs. 2005). et al. further studies are needed to investigate the anticariogenic properties of herbs. Yu. MICs were in excess of concentrations found in tea infusion. Muroi. The antioxidant capacities of herbal extracts ranked differently in .. 3. The anti-plaque activity of herbs was not determined in this study. was: green tea > osmanthus > mate > rosemary > rose > honeysuckle > duzhong > jiaogulan > lemongrass > jasmine > lavender > juhua (Table 3)..856. duzhong. whereas lavender had the lowest. Gugliucci. e. followed by osmanthus. it is difficult to directly compare the results from different studies due to difference in analytical methods. jiaogulan. 2001. sanguinis (Table 2). relatively little is known about its activity against periodontal pathogens such as Porphyromonas gingivalis and Prevotella intermedia.87 200 150 100 50 e 0 Ho R D M G R J O J L L J Herbs ne asm uhua aven ose sma uzh reen iaog emo ate ose ma ys ine ng nth ong tea ula de uc ras ry n r us kle s Fig. respectively (Tsai. After green tea leaves and osmanthus. oolong tea (semi-fermented tea leaves of C. Total phenolics and flavonoid contents of methanolic herbal extracts. the result. Tsai. and mate showed less inhibitory activity against S.Green tea extract was reported to be effective in the prevention of dental caries because of its antimicrobial and anti-plaque activity (Hamilton-Miller. Although the results herein indicate that a crude methanolic extract of rosemary suppresses the growth of cariogenic streptococci. respectively (Table 2). 2005. black tea. and osmanthus had no effect on the growth of S. a cup of green tea prepared with 2 g of tea leaves (the usual amount of a commercial tea bag) in 100 ml of infusion contains a total of 4.9 to 149 mg GAE/g extract. mutans up to 8 mg/ ml. mutans and S. After green tea leaves and osmanthus.001). / Food Chemistry 110 (2008) 859–864 teria S. Spieler. Contents of total phenolic compounds and flavonoids Total phenolics and total flavonoids in 12 herbal teas are shown in Fig. juhua. Muroi & Kubo. as shown in Table 2.001) were less well correlated. p < 0. 2001). rosemary (Tsai. Total antioxidant capacity Herbs may be good sources of natural antioxidants. mutans and S. exhibit bactericidal activity against S. The methanolic extracts from jasmine. sanguinis at a MIC of 1 mg/ml. 2007). In contrast. sobrinus at MICs of 16 and 4 mg/ml. The mechanism of antimicrobial action of these herbs has not been clearly elucidated. sanguinis.2. This study demonstrated that methanolic extract from rosemary also possessed antimicrobial activity against S. Total phenolics of herbal teas varied from 33.

5 ± 0.66 ± 0.07 3.81 Rutin 13.28 1. 1989) and nonpolar volatile flavour compounds (Kubo et al.65 13. lavender.2 ± 0. including several polar polyphenolic compounds (Sakanaka.79 93. lemongrass.4 0.79 1.001). the osmanthus-scented green tea.4 nd nd 1.90 6. these phenomena may be among reasons for the different observations using TEAC and DPPH assays.64 nd 0. .017 1.90 94.99 ± 0.425 ± 0.00 22.44 2.04 2.316 ± 0.18 ± 0.018). a Data are expressed as means ± SD.98 2.. A positive relationship was presented between total phenolics and total antioxidant capacity in 12 herbs.90 Myricetin 1. may be used as a good source of natural antioxidants. such as (À)-epicatechin and (À)-epigallocatechin. rose. 2005). We previously demonstrated that there was a good correlation between total phenolic and TEAC values in aqueous extracts from herbs. 1993). et al.006 4.928.80 ± 0. Silva.28 ± 0.-H.011 0. Chlorogenic acid was the predominant phenolic constituent in the extracts of honeysuckle. mutans and S.001). Cai and Wu (1996) reported that kaempferol Table 4 Phenolic compounds of methanolic herbal extracts Herbs Honeysuckle Jasmine Juhua Lavender Osmanthus Rose Duzhong Green tea Jiaogulan Lemongrass Mate Rosemary Chlorogenic acid 153 1.711.9 ± 1.089 0.07 26. and rosemary (Tsai.393. 2006). osmanthus flowers are also used to make osmanthus-scented jam.30 nd 0. *nd.24 863 Rank 6 10 12 11 2 5 7 1 8 9 3 4 *The concentration was 1 mg of methanolic extract per ml in DPPH radical reaction mixtures.240 ± 0. 1992).00 27. There is little information concerning the antioxidant activity of osmanthus.893. p = 0.44 0.34 ± 0.63 ± 0.. although both of them were based on the measurement of electron transfer propensity of antioxidant (Table 3).8 nd 23. Recently.12 4.001). TEAC (r = 0.37 1.313 ± 0. mutans. & Yamamoto.60 ± 0.4. expressed as DPPHÁ (r = 0.15 7.16 Each value is the mean (mg/g extract) of two replications.33 nd nd nd 2. Taniguchi.009 0.009 1.59 nd 0.25 nd 0. Kim. & Gloria. TEAC and DPPHÁ assays.05 4..90 Quercetin (mg/g of methanolic extract) 2.001).84 ± 0.05 1.04 0.58 nd 0.0 ± 1.15 ± 1.45 0. & Bernard.16 nd nd 1.T.01 nd 1.2 ± 1.31 23. 1985).65 12.007 0.01 0.3 0. Characterization of phenolic constituents The selected phenolics in the 12 tested herbal extracts were identified and are presented in Table 4.05 45.3 ± 0.076 Rank 6 12 9 11 2 5 8 1 7 10 3 4 ORAC Value (mmole TE/ g extract) 1. 3. Tsai. A significant relationship was also displayed between total flavonoids and total antioxidant capacity. Weil.5 ± 0.968. mutans up to 500 lg/ml (Muroi & Kubo. p < 0. Considerable variation was found in selected phenolic compounds of different herbs.13 nd 60.70 2. 2002).12 ± 0. juhua. p < 0.32 2. Flavanones and some dihydroflavonols inhibit the growth of S.13 2. and ORAC (r = 0.48 ± 1. Rutin was a major component in the extracts of rose.54 ± 0.045 1. Chlorogenic and caffeic acid inhibit the growth of enterobacteria and S. The antioxidant values correlated well with both total phenolics and total flavonoids contents.23 0.11 0.15 nd 1. TEAC (r = 0. future studies are needed to explore the role of active phytochemicals with respect to antioxidant activity. mutans (Almeida. The TEAC values were not strongly related to chemical structures or the number of electrons that an antioxidant can give away (Huang et al.0 10.54 4.4 1. Woods. and ORAC (r = 0.98 nd 6.002 0.06 0.27 0. sweet cake.20 89.. In Chinese cuisine. Yu. and soups. Numan.4 ± 1. mate and duzhong.8 21.64 ± 0. interest has increased considerably in finding naturally occurring antioxidants for use in foods or medicinal materials. p < 0.44 Caffeic acid 1. the polyphenolic compounds identified in green tea. jasmine.048 0. For lemongrass extract. sobrinus (Koo et al. all kinds of analyzed phenolic compounds were trace constituents.1 ± 1. Since the methanolic extracts from green tea leaves and osmanthus possess high antioxidant activity.341 ± 0.90 ± 0.9 6.92 ± 0. Many substances in green tea exhibit antibacterial activity against S.595 ± 0.18 0.11 3.27 10. such as green tea.045 1. p < 0.10 1.97 86.34 25.16 ± 0.65 3.274 ± 0.4 ± 0. 2007). p < 0. duzhong and mate. / Food Chemistry 110 (2008) 859–864 Table 3 In vitro free radical-scavenging activities of methanolic herbal extractsa Herbs DPPHÁ– scavenging Value (% quenched*) Honeysuckle Jasmine Juhua Lavender Osmanthus Rose Duzhong Green tea Jiaogulan Lemongrass Mate Rosemary LSD0.83 0.001). did not show any antibacterial activity against S.66 5.002 0. Osmanthus extract has been approved as a food additive and confirmed as GRAS in the USA (Oser.12 0.736.97 nd nd 0.48 ± 0.77 ± 0. which is familiar in Taiwan and served with tea bags or in restaurants.81 Kaempferol nd* 1. not detectable.1 2.80 3. Farah. expressed as DPPHÁ (r = 0. Tsai et al. Hence.88 2.31 Rank 6 10 7 12 2 3 8 1 11 9 4 5 TEAC Value (mmole TE/ g extract) 0.72 70.35 0.01 0. However.

Spieler. I. A. T. & Matsumoto. Y. These phenolic acids and flavonoids possess antioxidant activity (Kähkönen et al. S. 25(2). Matsudaira.. rosemary. Antimutagenic activity of flavonoids from Chrysanthemum morifolium. C. Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria. & Gimenez. Cho. J. B. A. Life Sciences. and Biochemistry. Food Chemistry. Archives of Pharmacal Research. 33(3).. Hampsch-Woodill. (1988). Quantification of polyphenolics and their antioxidant capacity in fresh plums. N. (1996). Lopez. sanguinis alone. D. C. (2003). A. A.864 T.. B. (1992). Journal of Agricultural and Food Chemistry. Preventive Medicine.. Kähkönen. & Choi. jiaogulan.). Antioxidant activity of plant extracts containing phenolic compounds. 45. C. 1999). (2005). & Himejima. W. Moon. Biochemical and Biophysical Research Communication. (2001). (2001). L. M. Y... 1302–1309. Yu. Carraminana.. A. 89. J.).. Jung. M. Journal of Agricultural and Food Chemistry. Antimicrobial Agents and Chemotherapy. XIV. Antibacterial substances in Japanese tea extract against Streptococcus mutans. mutans. M. A. Lee. (2005).... 108–117. C. Sasaki. Y. 97–107. Antioxidant activity and phenolic compounds in selected herbs. 491S–499S. 49. Y. Journal of Agricultural and Food Chemistry. & Mohamed.. and apigenin) content of edible tropical plants. & Kubo. Y. B. kaempferol. Clinical oral science (pp. 53.. (2006). Graham. L. Farah. M. 49(10). Journal of Agricultural and Food Chemistry. Reactive oxygen species scavenging activity of DuZong (Eucommia ulmoides Oliv. 2091–2099. U.. 987–990. Rota. 105. 3431–3436. & Hsieh. 348–352. Koo. M.. Kim. 71(9). Kim. Matsumura. B. F.. (2003).. 639–645. C. Antibacterial activity of Camellia sinensis extracts against dental caries.. / Food Chemistry 110 (2008) 859–864 Hardie. (1996). & Haider. (2005). J. Hypoglycemic effect of Du-Zong (Eucommia ulmoides Oliv. and myricetin had antimicrobial activity against S. Development and validation of an improved oxygen radical absorbance capacity assay using fluorescein as the fluorescent probe. Journal of Agricultural and Food Chemistry. In vitro inhibitory effects of rosemary extracts on growth and glucosyltransferase activity of Streptococcus sobrinus. R. Cai. American Journal of Clinical Nutrition.. K.. H. Keelung. Anti-cariogenic properties of tea (Camellia sinensis). P.. Kim. (2005). Food Technology... Y. S. H. 57(Suppl. et al. In conclusion. Woods. J. & Yamamoto. B. Roy..).. K... T. T.. H. The chemistry behind antioxidant capacity assays. Ou... 311–316.. Numan.. Comparison of NO-scavenging and NO-suppressing activities of different herbal teas with those of green tea. Oxford: Wright Press. (2004). 67(10). V. Osawa. Journal of Agricultural and Food Chemistry. A. Sobue. 49. Beneficial effects of green tea – A review. L. H. Craig... W. S. (1999). and mate possess antimicrobial activity against S. 22–28. Kim. Park. H. 67. Miean. Oser. Hopia. & Lee. 6509–6515. Tamesada. Journal of Agricultural and Food Chemistry. 334–350. C. G. S. 41361–41367.. Compounds from Syzygium aromaticum possessing growth inhibitory activity against oral pathogens. Agricultural and Biological Chemistry. Cury. A. Pihlaja.. (2003). green tea. M. luteolin. M. M. Theoduloz. S. Kum. The phenolic compounds in herbs are diverse and complex. Huang.. Rauha. 874–879. Journal of the American College of Nutrition. The results suggested that other unidentified phenolic components herein contributed to antioxidant capacities of herbal extracts and antimicrobial activity of rosemary against S. 181–187.. Journal of Dental Research. Hamilton-Miller. (1992). 11. Antiinflammatory activity of herbal medicines: Inhibition of nitric oxide production and tumor necrosis factor-alpha secretion in an activated macrophage-like cell line. Liao. H. M. L. 53. V. & Hamada. 5165–5170. Journal of Agricultural and Food Chemistry. M. Journal of Agricultural and Food Chemistry. J. R. 415–424. (2004). D. W. P. & Bowen. Park. Silva... & Herrera.. Although honeysuckle had the highest amount of chlorogenic acid among 12 tested herbs herein.. 39(11). J.. 53. References Almeida. Tsai. myricetin. Sakanaka. Free radical scavengers and antioxidants from lemongrass (Cymbopogon citrates (DC) Stapf. 67–71. Edgar. A. The dicaffeoylquinic acids derived from juhua showed potent DPPHÁ radical-scavenging activity (Kim & Lee. J. J. G. The American Journal of Chinese Medicine. (2001). M. T. 338–344. H. (2001). G. Tsai et al. Ghannadi. 1841–1856. J.. Caries inhibitory activity of cocao bean husk extract in in-vitro and animal experiments. 27. Phytochemistry.. Harris.. Food Chemistry. A. Park. & Lee. K. A. (1985). J. Journal of Biological Chemistry. Liepinsh.. and polyphenol chemistry..-H. Taiwan. intermedia. Journal of Medical Microbiology. Y. 70(Suppl. gingivalis. N. T. it possessed moderate antioxidant activity and no inhibitory effect on the growth of S. (2004). & Andriantsitohaina. Rodrigues.. Bixby. K. 41. Tsai.. & Hisama..... Journal of Agricultural and Food Chemistry.. honeysuckle. E. Journal of Agriculture and Food Chemistry. 51. 299–302. Kawabata. Current Medical Chemistry. 3106–3112. (2006). Bioscience. 103. Inhibitory effects of apple polyphenols and related compounds on cariogenic factors of mutans streptococci. Planta Medica. T. sanguinis. A. L. Health-promoting properties of common herbs. (1999)... & Oliveria Cordeiro. A novel insulin-releasing substance. R.. Flavonoid (myricetin. Chlorogenic acid and caffeic acid are well-known plant antioxidants and show a strong effect toward DPPHÁ (Cheel et al. The crude methanolic extracts from jasmine... K. & Lai. J.. (2002). A.. S. GRAS substances. C. Di. J. S. A universal HPLC method for the determination of phenolic acids in compound herbal medicines. 2005).. A.. Gugliucci. B. (2001). Kanda. Weil. Y. 774–778.. J. and P. 21(3). Journal of Agricultural and Food Chemistry. Burillo. S. Future investigations should examine the role of herbal teas in a wider range of bacteria. 79–99. In M. the present study shows that crude methanolic extract of rosemary can inhibit the growth of the cariogenic bacteria. Rasheed. 8738–8743.. C.. M.. H. Filip. Muroi. Menini. 53. Effects of compounds found in propolis on Streptococcus mutans growth and on glucosyltransferase activity. T.. R.. In vitro antimicrobial activity of essential oils from aromatic plants against selected food borne pathogens. Larson. Park. T. 72. Antimicrobial and chemopreventive properties of herbs and spices. Miyazawa. & Prior. 2307–2311. including Gram-negative species associated with gingivitis and other forms of periodontal diseases. O... Kujala. (1998). P. H. Zheng.. H. (1989). Osaka. 245–248. 48. & Wang. Li. K. Journal of Food Protection. Park. Pharmacological Reports. (2005). I. S. M. C. In summary. Taniguchi.. & Gloria.. Journal of Agricultural and Food Chemistry. Biotechnology. mutans. & Heinonen.. (2005). R. C.. 50. Kim. E. Park. M. Diabetes Research and Clinical Practice. H. 199–212). & Gugliucci. consumption. 6624–6629. & Wu. A. mutans and the periodontal pathogens. Yasuda. S. P. G.. Tsai.) leaves in streptozotocin-induced diabetes rats. K. C. O. lemongrass. 224(2). Chun. S. Antimicrobial activity of green tea flavor components and their combination effects. & Bernard. & Prior. 2511–2517.... N. H. P. C. The antioxidants of higher plants. Journal of Natural Products. Meghji (Eds. G. Acknowledgement This work was supported by the Grant CMRPG 250301 from the Keelung Chang Gung Memorial Hospital. Journal of Ethnopharmacology. Yen. L. R.. J. from the plant Gynostemma pentaphyllum.. 1252–1256. S.. 871–876.. Kubo. & Ferraro.. Fitoterapia. D. S. Thuan. Yanagida.. M. Tanabe.). quercetin. Jornvall. S. Synergistic effects of streptococcal glucosyltransferase on adhesive biofilm formation. Rosalen. R. I. Y. M. (1998). 83(11). these results suggest that herbs may have potential application in preventing dental caries and other health problems associated with free radical-mediated damage.. a cariogenic bacterium. 279. Fujiwara. K. K. 67. Giberti. J. Recent progress in the consideration of flavoring ingredients under the Food Additives amendment. Muroi. (2005). J.. 54. Archives of Oral Biology. & Ho. & Schuller-Levis. The microbiology of dental caries and periodontal disease. L. & Ho. M. Tsai. . D. 48(11). H. Curin. (2004). M. M. 2005). A. J. Ou.. A. M. Anti-inflammatory and analgesic properties of the leaf extracts and essential oil of Lavandula angustifolia Mill. (2000). 59. 77. phanoside. Green tea composition. (2000). H. (1993). & Schmeda-Hirschmann. J. B. & Liu. mutans and S. S.. D.. Vuorela. P. 4619–4626. D. H. such as quercetin. H. 47. & Sharif. 1451–1460. 5666–5671. 969–978. Y. Hoa. S. M.. Kim. H. T. J. Combination effects of antibacterial compounds in green tea flavor against Streptococcus mutans. (2005). kaempferol and their glycosides. Y. Ooshima. (2007). Wang.) and its active compounds. & S. 3954–3962. M. Antioxidant effects of Ilex paraguariensis: Induction of decreased oxidability of human LDL in vivo.. K.. Polyphenols as potential therapeutical agents against cardiovascular diseases. A.. Hajhashmi. the main flavonoid constituents are flavonol aglycones. (2000). Cabrera. P. D. Ilex paraguariensis extracts are potent inhibitors of nitrosative stress: A comparative study with green tea and wines using a protein nitration model and mammalian cell cytotoxicity. G.. C. M. duzhong. Phan. 1102–1105.. Identification of new dicaffeoylquinic acids from Chrysanthemum morifolium and their antioxidant activities. S. L. 21. Wen. Norberg. 46(5). Y. This study also reveals that osmanthus may be an effective potential source of natural antioxidants.. Artacho.. Coussio. Phenolic compounds in seven South American Ilex species. K. S. In many herbs. Cheel. E. H. 40. J. (2007). 345–358.