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Journal of Pharmacognosy and Phytochemistry Evaluation Of ISSN 2278- 4136 ZDB-Number: 2668735-5 IC Journal No: 8192 Volume 2 Issue 1 Online Available at www.phytojournal.com Journal of Pharmacognosy and Phytochemistry Evaluation of Antioxidant and Antimicrobial Activities from 28 Chinese Herbal Medicines Cheng-Hong Yang1, Hsueh-Wei Chang2,3, Ho-Yang Lin4 , Li-Yeh Chuang4* 1. Department of Electronic Engineering, National Kaohsiung University of Applied Sciences, Kaohsiung, 807, Taiwan. 2. Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung, 807, Taiwan. 3. Cancer Center, Kaohsiung Medical University Hospital; Kaohsiung Medical University, Kaohsiung, 807 Taiwan. 4. Institute of Biotechnology and Chemical Engineering, I-Shou University, 840 Taiwan [E-mail address: [email protected]] Background: Many Chinese medicines have been reported to exhibit high antimicrobial and antioxidantactivities. In this study, 28 traditional Chinese herbal medicines were tested for their antioxidant and antibacterial activities. Materials and Methods: Total flavonoid content of the ethanol extracts were determined by a colorimetric method. Total phenol content was estimated as gallic acid equivalents. The antioxidant activities of the extracts were evaluated by various antioxidant assays, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging, trolox equivalent antioxidant capacity (TEAC) and reducing power. In addition to the antioxidant activity, the antimicrobial assay was measured as well. Six strains of the clinical antibiotic resistant pathogens including Escherichia coli, Staphylococcus aureus, methicine resistant Staphylococcus aureus (MRSA), ornithine resistant Staphylococcus aureus (ORSA), Pseudomonas aeruginosa, and Acinetobacter baumannii were used in the tests. Results: Among the extracts screened, Artemisia vulgaris and Sanguisorba officinalis showed the best antioxidant performance in all the tested methods.In vitro antibacterial activity, the extracts of Paeonia suffruticosa and Cinnamomum cassia displayed a broad antimicrobial spectrum and significantantimicrobial activities, with inhibition zones between 14-28 mm, minimum inhibitory concentration (MIC) between 0.9-5.2 mg/mL, and minimum bactericidal concentration (MBC) between 1.0-5.7 mg/mL. Conclusion: These data suggest that Artemisia vulgaris and Sanguisorba officinalis may be effective potential sources of natural antioxidants, and Paeonia suffruticosa and Cinnamomum cassia are potent inhibitors of antibiotic resistant pathogens. Keyword: Chinese Herbal Medicines, Antioxidant Activity, Antibacterial Activity 1. Introduction antioxidants possess a wide range of biological It is a well-known fact that oxidationdamages activities, including inhibition of reactive oxygen various biological substances and subsequently species (ROS) generation, direct or indirect causes many diseases, such as cancer, liver scavenging of free radicals, and alteration of disease, Alzheimer’s disease, premature aging, intracellular redox reactions. As a result, arthritis, inflammation, diabetes, Parkinson’s antioxidants provide protection to living disease, and atherosclerosis[1]. Many previous organisms from damage caused by uncontrolled literature reports indicated that natural production of reactive oxygen species and the Vol. 2 No. 1 2013 www.phytojournal.com Page | 294 Journal of Pharmacognosy and Phytochemistry concomitant lipid per-oxidation, protein damage flavonoids that may occur in all parts of plants and DNA strand breakage. Antioxidants, which such as fruits, vegetables, nuts, seeds, leaves, can quench reactive free radicals, can prevent the roots and barks. Many of these bioactive oxidation of other molecules and may thereby compounds possess antioxidant, antimutagentic, prevent degenerative diseases[2-3]. anticarcinogenic, antibacterial or antiviral Antibiotics provide the main basis for the therapy activities to a greater or lesser extent[4]. of microbial infections. However, the high Considering the rich diversity of plants, it is genetic variability of these pathogenic microbes expected that screening and scientific evaluation enables them to rapidly develop antibiotic of plant extracts for the antioxidant and resistance. In recent years, development of multi- antimicrobial activities may provide new drug resistance in the pathogenic microbes has antioxidant and antimicrobial substitutes[5]. created major clinical problems in the treatment Chinese herbal medicines extracts obtained from of infectious diseases and other problems such as many plants have recently gained popularity and toxicity of certain antimicrobial drugs on the host scientific interest. Many Chinese herbal tissue, which triggered interest in the search of medicines have been used for different purposes, new antimicrobial substances/drugs of plant such as food, drugs and perfumery, not only as origin. flavoring agents, but also as folk medicine and Phytomedicines have shown great promise in the food preservatives. Researchers have been treatment of various diseases including interested in biologically active compounds microorganism infections. Single and polyherbal isolated from Chinese medicines for the preparations have been used throughout human antioxidant and antimicrobial activities. The main history for the treatment of various types of objectives of this study were: (i) to investigate the illness. The medicinal properties of plants have antimicrobial activity of the ethanol extracts from been investigated, in the light of recent scientific 28 Chinese medicines by disc diffusion method, developments, throughout the world due to their minimuminhibitory concentration and time potent pharmacological activities and economic killing curve against the clinical drug resistance viability. A great number of aromatic, spicy, pathogens; (ii) to evaluate the antioxidant medicinal and other plants contain chemical capacity of the ethanol extracts by free radical compounds, exhibiting antioxidant and scavenging effect and reducing power. Besides, antimicrobial properties. Sources of natural active this study also determined the contents of substances are primarily, plant phenolics and flavonoids and phenolics of the extracts. Fig 1: Effect of the extracts from Chinese herbal medicines on growth of Acinetobacter baumannii strain. Vol. 2 No. 1 2013 www.phytojournal.com Page | 295 Journal of Pharmacognosy and Phytochemistry 2. Materials and Methods Polygonatum modoratum, Psoralea corylifolia, 2.1 Plant Materials Rheum palmatum, Salvia miltiorrhiz, Chinese herbal medicines used in this study were Sanguisorba officinalis, Scrophularia selected based on their usage as folk medicine, as mingpeonsis, Senecio scandens, Sophora well as indications of the presence of compounds japonica, and Vladimiria souliei. Plant materials with antimicrobial properties. 28 Chinese herbal were purchased from local folk medicinal dealers. medicines have been used for screening, including Acanthopanax spinosus, Acorus 2.2 Preparation of Crude Extracts calamus, Androgra phispaniculata, Artemisia The dried Chinese herbal medicines (100 g) were vulgaris, Carpesium abrotanoides, extracted with 500 mL of 95% ethanol overnight Chrysanthemum indicumi, Cinnamomum cassia, by shaking in an incubator at 200 rpm and 37℃. Coptis chinensis, Cremastra variabilis, After filtration, the extracts were concentrated Cynanchum bungei, Euphorbia lathyris, under reduced pressure by a vacuum rotary Fritillaria thunbergii, Houttuynia cordata, Isatis evaporator. The dried extracts were stored at 4℃ tinctoria, Matteuccia struthiopteris, Melia until the assay. azedarach, Ompalia lapidecene, Paeonia suffruticosa, Phellodendron amurense, Fig 2: Effect of the extracts from Chinese herbal medicines on growth of Pseudomonas aeruginosa strain. 2.3 Assay For Total Phenolics acid calibration curve was Y=0.0098·X- 0.0172 The total phenolic content (TPC) of extracts was (where X = concentration of gallic acid determined using the Folin–Ciocalteu reagent[6]. equivalents (GAE) expressed as milligrams of Briefly, 0.2 mL of each extract was shaken for 1 GAE per gram of dried extract; Y = measured min with 1 mL of 0.5 MFolin–Ciocalteu reagent. absorbance), and the correlation coefficient was After 4-8 min mixture was shaken and added R2=0.9965 1mL of 75 g/L Sodium carbonate, the mixture was shaken once again for 0.5 min. After 2 h, the 2.4 Assay For Total Flavonoids absorbance was read on the UV/Visible The total flavonoid content was established in the spectrophotometer at 760 nm. The TPC was reaction with aluminum chloride using the assessed by plotting the gallic acid calibration methods of Chang[7]. Briefly, 1 mL of each curve (from 10 to 200 mg/L) and expressed as extract was shaken for 1 min and added 0.1 mL milligrams of galic acid equivalents (GAE) per of 10% alumminium chloride, 0.1 mL 1M gram of dried extract. The equation for the gallic potassium acetate and 3.8 mL of methanol. After Vol. 2 No. 1 2013 www.phytojournal.com Page | 296 Journal of Pharmacognosy and Phytochemistry 40 min at room temperature, the absorbance was Y=0.0101. X- 0.0632 (where X = concentration read on the UV/Visible spectrophotometer at 415 of Quercetin equivalents expressed as milligrams nm. The flavonoids was assessed by plotting the of Quercetin per gram of dried extract; Y = Quercetin calibration curve (from 10 to 200 measured absorbance), and the correlation mg/L) and expressed as milligrams of Quercetin coefficient was R2=0.9967 equivalents per gram of dried extract. The equation for the Quercetin
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