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Antioxidant and Antimicrobial Activity of Zingiberaceae Plants in Taiwan

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Antioxidant and Antimicrobial Activity of Zingiberaceae Plants in Taiwan Plant Foods Hum Nutr (2008) 63:15–20 DOI 10.1007/s11130-007-0063-7 ORIGINAL PAPER Antioxidant and Antimicrobial Activity of Zingiberaceae Plants in Taiwan I-Nan Chen & Chen-Chin Chang & Chang-Chai Ng & Chung-Yi Wang & Yuan-Tay Shyu & Tsu-Liang Chang Published online: 20 December 2007 # Springer Science + Business Media, LLC 2007 Abstract The rhizomes of the Zingiberaceae family are a Keywords Antimicrobial . Antioxidant . Ginger . vegetable widely used in many Asian countries, and their Reducing power . Zingiberaceae medicinal functions have been broadly discussed and accepted in many traditional recipes. In this study, 18 species of five genus of Zingiberaceae plants from Taiwan area were Introduction collected and analyzed for their functional properties. Methanolic extracts of the plants were analyzed for their The Zingiberaceous plants are characterized by their total phenol compounds, α,α-diphenyl-β-picrylhydrazyl tuberous or non-tuberous rhizomes, which have strong (DPPH) scavenging activity, and reducing power. Antimi- aromatic and medicinal properties. It is commonly known crobial activity of these samples was also determined. The as ginger, and exists in about 50 genera and 1,300 species results showed that the total phenol compounds of the Alpinia worldwide, distributed mainly in South and Southeast Asia genus averaged 17, 30 mg/g for Curcumas, and the highest, [1]. Turmeric of Zingiberaceous plants in powder form is 36.5 mg/g for Vanoverberghia sasakiana. Antioxidant widely applied as a food additive in many Asian countries. performances were best observed in Vanoverberghia and Medicinal functions for treatment of diseases such as Hedychium, both 89%, and DPPH scavenging activity diarrhea, coryza, dermatosis disorders and rheumatism are followed similar trends. Particularly, Zingiber oligophyllum, also widely mentioned in traditional remedies [2–5]. considered as a traditional medicinal plant used in Taiwan Zingiberaceae plants contain many essential oils, including exhibited low DPPH scavenging activity and reducing terpenes, alcohols, ketones, flavonoids, carotenoids and phy- power. Most Zingiberaceae plant extracts exhibited antimi- toestrogens [6–8]. For instance, the water extract of Zingiber crobial activity against all tested food microorganisms. officinale exhibits 6-Gingerol, and is mostly found in the Hedychium and Vanoverberghia, did not show antimicrobial rhizome in concentrations of 130–7,138 ppm. The major activities on Escherichia coli and Vibrio parahaemolyticus. functional compound in Curcuma turmeric is curcumin, This study is a positive demonstration of the utility of which has concentrations as high as 38,000 ppm in certain screening Taiwan’s endemic Zingiberaceous plants for their species [8]. Less-polar constituents including curcuminoids, food and medicinal uses. kava pyrones and gingerols isolated from Zingiberaceous plants, which have been reported for their biological activities : : : : I.-N. Chen C.-C. Ng C.-Y. Wang Y.-T. Shyu in antifungal, antioxidant, insecticidal, and anti-inflammatory T.-L. Chang (*) activities [9–11] are particularly important and relevant for Department of Horticulture, National Taiwan University, these applications. 140 Keelung Rd. Section 4, Taipei 10600 Taiwan, Republic of China This study reveals the bioactive and medicinal potentials e-mail: [email protected] of ginger species in Taiwan. Six genera and 25 species of Zingiberaceae plants are native to Taiwan. Among these, C.-C. Chang Alpinia is the largest genus, with 14 species recorded so far Department of Living Sciences, Tainan University of Technology, 529 Jhongjheng Rd., Yongkang, [12]. The antioxidant and antimicrobial properties of Tainan 71002 Taiwan, Republic of China Taiwan’s endemic and imported Zingiberaceae plants have 16 Plant Foods Hum Nutr (2008) 63:15–20 not yet been thoroughly explored [7]. This work evaluated Antioxidant Activity Determination the antioxidant and antimicrobial properties of 18 Taiwan endemic species collected from all over the island. The The antioxidant capacity of samples was measured by the concentrations of phenolic compounds, antioxidant capac- method described by Miller and Rice-Evans, and Arnao et al. ity, α,α-diphenyl-β-picrylhydrazyl (DPPH) scavenging [14, 15] with little modifications. Peroxidase (4.4 units/ml, activity and reducing power of the samples were deter- Sigma, MO, USA), H2O2 (50 μM, Merck, Germany), 2, mined. The activities of ginger rhizomes against different 2-azino-bis (3-ethylbenz-thiazoline-6-sulphonic acid) food-related microorganism were also measured. This [100 μM, Sigma, MO, USA] and distilled water (1 ml) investigation evaluates the potential medicinal applications were mixed and kept in dark for 1 h for reaction. One ml of of Taiwan’s endemic Zingiberaceae plants. plant extract was subsequently added and determined by absorbance at 734 nm. The antioxidant capacity was calculated by the following formula: Materials and Methods Antioxidant activity (%) = [1−(A734 nm, sample/ A734 nm, control)] × 100 Rhizome Collection DPPH Free Radical Scavenging Activity Test Plant materials were collected and purchased from the Taiwan Endemic Species Research Institute, the Council of The capacity of Zingiberaceae to remove 1,1-diphenyl-2- Agriculture, traditional markets, and local growers from picryl-hydrazyl radical (Sigma, MO, USA) was determined whole Taiwan Island. The rhizomes were removed from the by the method described by Shimada et al. [16]. Briefly, plant and cleaned for use. 1 ml of methanolic extract and 5 ml of freshly prepared 0.1 mM DPPH methanolic solution were thoroughly mixed Sample Treatment and Extraction and kept in the dark for 60 min. The absorbance of the reaction mixture at 517 nm was measured with a spectro- Peeled and washed samples were further chopped into 2× photometer. The blank was prepared by replacing the 2 cm dice and dried with freeze-dryer (Freeze dryer Alpha extract with methanol (1 ml). The percentage of free radical 1-2/LD-2, Vacuum pump RZ-5, Christ, Germany) for 48 h. scavenging activity was calculated as follows: Dried samples were subsequently milled using commercial Scavenging activity (%) = [1−(A517 nm, sample/ hand-carry milling machine. For methanol extraction, A517 nm, blank)] × 100 sample powders (1.25, 2.5, 3.75, or 5 g) were mixed with 20 ml methanol (Sigma, St. Louis, MO, USA) in a rotary Reducing Power Test shaker for 12 h. The mixtures were then filtered (Whatman No.1, Middlesex, England). The filtrate was then concen- Reducing power was measured according to the method trated in a rotary evaporator (Büchi R-210, Flawil, Switzer- described by Duh and Yen [17]. One ml of plant extract, land) until dried. The weight of extract was then measured phosphate buffer (0.2 M, pH 6.6, 0.5 ml), and potassium and recorded. The yield of methanolic extract was hexacyanoferrate solution (1% v/w, 2.5 ml; both from expressed as milligram of extract per gram of ginger fresh Merck) were placed in a test tube and heated at 50 °C for weight, calculated as follows: 20 min. The tube was cooled on ice and 0.5 ml 10% Yield = weight of Rotavapor dried sample (mg)/fresh trichloroacetic acid (Merck) were added. After centrifuga- sample (g) × 100% tion at 3,000×g for 10 min, a 1 ml of aliquot supernatant was mixed with 1 ml distilled water and 0.1 ml ferric Total Phenolics Determination chloride (0.1%) and reacted for 10 min. Finally, the absorbance at 700 nm was measured, increased absorbance The total phenolics content of ginger extracts was deter- of the reaction mixture indicated increased reducing power. mined by the Folin–Ciocalteau method [13]. Briefly, 0.5 ml diluted extract solution was shaken for 1 min with 100 μlof Antimicrobial Activity Folin–Ciocalteau reagent and 6 ml of distilled water. The mixture was shaken and 2 ml of 15% Na2CO3 were added To screen the activity of methanolic extracts of Zingiberaceae and shaken once again for 30 s. Finally, the solution was plants against food related microorganism, a filter paper disc brought up to 10 ml by adding distilled water. After 1.5 h, method was used as described by Gülçin et al. [18]. the absorbance at 750 nm was evaluated using a spectro- Escherichia coli Bioresources Collection and Research photometer (Beckman Coulter DU-640, CA, USA). The Center (BCRC) 10675, Salmonella enterica BCRC 10242, results were expressed as gallic acid equivalents. Staphylococcus aureus spp. aureus BCRC 10451 and Vibrio Plant Foods Hum Nutr (2008) 63:15–20 17 parahaemolyticus BCRC 12863 were purchased from BCRC, Table 1. Extraction yield ranged from lowest 41 mg/g Republic of China. E. coli stock cultures were maintained on (Alpinia pricei Hayata) to highest 127 mg/g (Alpinia potato dextrose agar (Difco, Detroit, MI, USA) and the other zerumbet (Pers.) Burtt & Smith), notably Vanoverberghia three were kept on nutrient agar (Difco, Detroit, MI, USA) sasakiana, the endemic species Taiwan has second high in according to the culture manual. Culture concentrations of 1× extraction rate of 177 mg/g. Average extraction rate was 108 colony-forming unit/ml were plated on their respective found to be 0.084% (84 mg/g). Habsah et al. [6]has agar plates. Sterile filter paper discs with 6 mm diameter displayed the methanolic extraction rate of Alpinia, Costus (Whatman) were immersed with 500 μl of Zingiberaceae and Zingiber, which ranged 0.09–0.57% in various samples. extract. After thorough absorption, they were then placed on In general, freeze drying preserves more reducing power of agar plate for cultivation. Salmonella plates were incubated at plant extract, such as yam (Dioscorea sp.) extract as 30 °C, while E. coli, Staphylococcus and Vibrio plates were compared to hot-air drying and drum drying [20]. The color kept at 37 °C for 72 h. The results were recorded by and appearance of our vacuum-dried samples remained measuring the zones of growth inhibition surrounding the unchanged and intact. V. sasakiana showed a 36.5 mg/g of disc. Clear inhibition zones around the discs indicated the total phenol content (Table 1), highest in Taiwan endemic presence of antimicrobial activity.
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