Nghiên cứu canh tác và các hoạt động sinh học của các thảo dược long nha thảo; hạn liên thảo, sương sáo, điền cơ hoàng, cam thảo đất và sả dịu

國國國國國國國國國國國國國國國國國國 Department of Tropical Agriculture and International Cooperation National Pingtung University of Science and Technology 國國國國國國 Ph.D Dissertation 國國 (國國國國國國國國國國國國國國國國國國國國國國)國國 國國國國國國國國 Study in Cultivation and Bioactivities of Herbs Agrimonia pilosa, Eclipta alba, Mesona procumbens, Hypericum japonicum, Scoparia dulcis, and Cymbopogon flexuosus 國國國國 Advisors: 國國國國國(Ming-Chang Wu, Ph.D.) 國國國國國(Horng-Liang Lay, Ph.D.) 國國國 Student: 國國國 (Le Quang Ung) 國國國國 108 國 國 15 國 June 15, 2019 國國 國國國P10422023 國國國國: 國國 (國國國國國國國國國國國國國國國國國國國國國國)國國 國國國國國國國國 國國國國138 國 國國國國國國國國國國國國國 國國國國國國國國國國國國國國國國 國國國國國國國國國107 國國國國 國國 國國國國國國國國 國國國: 國國國 國國國國: 國國國 國國 國國國 國國 國國國國國國國 國國國國國國國國國國國國國國國國國國國國國國國國國國國國國 國國國國國國國國國國國國國國國國國國國國國國國國(VC)國國國國國 國國國國Agrimonia pilosa Ledeb.國國國國國國國國國國國國國國國國國 國國國國國國國國國國國國國國 (Agrimonia pilosa Ledeb.)國國國國國國 國國國國國國 HepG2國國國國國國國國國 A549國國國國國國國國國國 國 國 國 國 國 國 國 國 國 Eclipta alba Hassk 國 國 國 國 (Mesona procumbens Hemsl.) 國 國 國 國 國 Hypericum japonicum Thunb 國 國 國 國 國 (Scoparia dulcis L.) 國 國 國 國 國 (Cymbopogon flexuosus (Nees ex Steud.) W Watson) 國國國國國國國國國國國國國國國國國國國國國國國國國 國國國國國國 國國國(ABTS)國國國國國國國國(DPPH)國國國國國國國國國國國國國 國國國國國國國國國國國國國國國國國 國國國國國國國國國國國國國國 國國國國國國HPLC國國國國國國國國國國國Agrimonia pilosa Ledeb.國國 國國國Eclipta alba Hassk.國國國國(Mesona procumbens Hemsl.) 國 國國 國 國Hypericum japonicum Thunb.國國 國國國 (Scoparia dulcis L.) 國國國 國 國 國 國 國 國 國 國 國 國 國 國 (MS) 國 國 國 國 國 (NMR) 國 國 國 國 國 國 (Cymbopogon flexuosus (Nees ex Steud.) W Watson) 國國國國國國國國國 I 國國國國國國國國國國國國國國國國國國國國國 bcl-2國bax國bad國 caspase-3國caspase-9 國 p53 國國國 國國國國國國國國國國國國國1國國國國國國國國國國國國國國 國Agrimonia pilosa Ledeb.國國國國國國國國國國國國國國國 pH 國國 6.9 國 7.4 國國國國國國 (EC)國 0.5-0.6mS cm-1 國國國國國國國國國國國國國 國國國(VC)國國國國國國國國國 12.5 + 16.875 ton ha-1 國國國國國國國國 國國國國國國國國國國國國國國國國國國國國國國國國國國國國國國國 國 國 國 國 國 國國 國 國 國 國 國 國國 國 國 國國 國 國國 2國國 國國 國 國國 國Agrimonia pilosa Ledeb.國國國國國國國國國 4-國國國國國國國國國國 國國國國國國國國國國國國 國國國國國國國國國國國國國國國國國國國 國國國國國國國國 A549 國國國國國國國國國國國國國國國國3國國國國 國國Eclipta alba Hassk.國國國 國 國 國 國 國 國 國 國 國 國 4- 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 (Mesona procumbens Hemsl.) 國國國國國國國國國國國國 7-國國國國國國國國國國國國國國國國國國 國國國國國國國國國國國國國Eclipta alba Hassk.國國國國國國國國國國 國國國國國國國(DPPH)國國國國國國國(ABTS)國國國國國國國國國國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 國 Hypericum japonicum Thunb.國國 國國國國國國國國國國國國國國國國國國國國國國國國國國 國 (Scoparia dulcis L.)國國國國國國國國國 國國國國國國國Hypericum japonicum Thunb.國國國國國國國國國國國國國國國國國國國國國國國國 國5國國1,3-O-國-E-國國國國國國SA3國國 1-O-p-國國國-3-O-國國國國 國國SA4國國國國國國國國 國(Cymbopogon flexuosus (Nees ex Steud.) W Watson) 國國國國國國國國國國國國 SA3 國國國國國國國國國國國國國 國國國國國 國國國國國國國國國國國國國國國國國國國國國國國國國國國國國 國國國國國國國國國國國國國國國國國國國國國國國國國國國國國國國 國 Agrimonia pilosa Ledeb 國 國 國 國 國 Eclipta alba Hassk 國 國 國 國 國 國 Hypericum japonicum Thunb 國 國 國 國 國 國 (Cymbopogon flexuosus (Nees ex Steud.) W Watson) 國國國國國國國國國國國國國國國國 國國國國國國國國國國國國國國國國國國國國國國 II Abstract Student ID: P10422023 Title of Dissertation: Study in Cultivation and Bioactivities of Herbs Agrimonia pilosa, Eclipta alba, Mesona procumbens, Hypericum japonicum, Scoparia dulcis, and Cymbopogon flexuosus Total Page: 138 pages Name of Institute: Department of Tropical Agricultural and International Cooperation, National Pingtung University of Science and Technology Graduate Date: June15, 2019 Name of Student: Le Quang Ung Degree Conferred: Doctoral Degree Advisors: Ming-Chang Wu, Ph.D Horng-Liang Lay, Ph.D The content of abstract in this dissertation: Six herbs chosen in this work have received considerable attention as natural products for their applications in traditional medicine The study consists of five studies being conducted under in vitro conditions The first study aims to investigate effects of vermicompost and hog manure on growth, yield, chemical composition of Agrimonia pilosa Ledeb Four other studies aim to investigate antioxidant activities, HepG2 (human liver cancer cell line) and A549 (adenocarcinomic human alveolar basal epithelial cell line) growth inhibitory effect from extracts of Agrimonia pilosa Ledeb Eclipta alba Hassk Mesona procumbens Hemsl Hypericum japonicum Thunb Scoparia dulcis L.and Cymbopogon flexuosus (Nees ex Steud.) W Watson Extracts of ethanol were chosen for the assays Total phenolic and flavonoid content and the antioxidant activities comprising ABTS+, DPPH, hydroxyl radical scavenging systems were determined Lactate dehydrogenase release in medium was also evaluated in some experiments High performance liquid chromatography (HPLC) method was used to analysize phenolic compounds from Agrimonia pilosa Ledeb Eclipta alba Hassk Mesona procumbens Hemsl Hypericum japonicum Thunb Scoparia dulcis L Chemical structural characterization from the aerial parts of Cymbopogon flexuosus was established by MS and NMR spectra techniques.The apoptotic factors bcl-2, bax, bad, caspase-3 and caspase-9, p53 were analyzed by western blotting assays The results showed that, for the first study (1), application of ogarnic fertilizers enhanced the morphological growth, yield, chemical composition of Agrimonia pilosa Ledeb The pH between 6.9 and 7.4, EC from 0.5-0.6 mS cm-1 are optimal for this herbal growth A mixture of the Vermicompost and Hog manure made with the rate of 12.5 + 16.875 ton ha-1 produced best parameters This result suggests that using of organic fertilizers could completely enhance the morphological growth, yield, chemical composition of other herbs and futher detailed investigations are in progress In the second study (2), two individual phenolic compounds consisting of 4-hydroxybenzoic acid and p-coumaric acid were firstly identified from Agrimonia pilosa Ledeb The root extract exhibited higher antioxidant and A549 inhibitory capacity compared to the aerial part extract In the third study (3), the 4hydroxybenzoic acid, caffeic acid, p-coumaric acid and rosmarinic acid were the identified compounds in Eclipta alba Hassk The 7-hydroxycoumarin, ferulic acid and rutin being new compounds and two known compounds were identidied in the Mesona procumbens Hemsl The higher levels of DPPH and ABTS+ radical scavenging and anticancer activities were detected from Eclipta alba Hassk In the fourth study (4), the chlorogenic acid and rosmarinic acid were the identified compounds in the Hypericum japonicum Thunb The rutin and rosmarinic acid were newly identified compound from the Scoparia dulcis L The higher levels of antioxidant and anticancer capacity were detected from Hypericum japonicum Thunb The last study (5), 1,3-O-di-E-caffeoylglycerol (SA3) and 1-O-p-coumaroyl-3-O- caffeoylglycerol (SA4) were firstly isolated from Cymbopogon flexuosus (Nees ex Steud.) W Watson The SA3 compound showed the highest antioxidant and anticancer potent The information obtained from this work will likely contribute to the development of novel natural medicine for cancer diseasers It will also provide new insights for the futher understanding of the functions and molecular mechanism of herbs An insightful investigation to establish and manufacture capsules used as function and supplemental food from Agrimonia pilosa Ledeb Eclipta alba Hassk Hepericum japonicum Thunb Cymbopogon flexuosus (Nees ex Steud.) W Watson was recommended Keywords: anticancer, Agrimonia pilosa Ledeb, Cymbopogon flexuosus (Nees ex Steud.) W Watson, Eclipta alba Hassk, Hypericum japonicum Thunb Acknowledgements I would like to express sincere thanks to my two advisors, Professor Dr Ming-Chang Wu and Professor Dr Horng-Liang Lay, for their invaluable guidance on my research I greatly appreciate their intellectual instructions, kindness, and supports during my Ph.D research program I would like to thank the advisory committee members for their valuable comments and constructive suggestions for the successful completion of this research work I am grateful to Taiwan Scholarship (Ministry of Education) for budget support for my Ph.D program in Taiwan I would also like to thank NPUST, DTAIC for offering me this good opportunity to pursue my doctoral study I would like to thank the personnel of the Department of Tropical Agriculture and International Cooperation; Office of International Affair, Chinese Herbal Medicine and FP201 Laboratory for their help during this study program Lastly, I would like to thank my parents for their confidence and love; my colleague, my friends for their encouragement and affection and especially my wife who was always there with wisdom, inspiration and cheer Table of Contents 國 國 I Abstract III Acknowledgements VI Table of Contents VII List of Tables XIII List of Figures XV List of Abbreviation XVIII Chapter Introduction 1.1 Research background 1.2 Research objectives Chapter Literature Review 2.1 Herbs used in the study 2.1.1 Agrimonia pilosa Ledeb (AL) 2.1.1.1 Botany 2.1.1.2 Chemical composition in AL 2.1.1.3 Pharmacological activities and the use of AL in traditional medicine 2.1.2 Eclipta alba Hassk (EH) 2.1.2.1 Botany 2.1.2.2 Chemical composition in EH 2.1.2.3 Pharmacological activities and the use of EH in traditional medicine 2.1.3 Mesona procumbens Hemsl (MH) 2.1.3.1Botany VII 2.1.3.2 Chemical composition in MH 2.1.3.3 Pharmacological activities and the use of MH in traditional medicine 2.1.4 Hypericum japonicum Thunb (HT) 2.1.4.1 Botany 2.1.4.2 Chemical composition in HT 10 2.1.4.3 Pharmacological activities and the use of HT in traditional medicine 10 2.1.5 Scoparia dulcis L (SL) 10 2.1.5.1 Botany 10 2.1.5.2 Chemical composition in SL 11 2.1.5.3 Pharmacological activities and the use of SL in traditional medicine 11 2.1.6 Cymbopogon flexuosus (Nees ex Steud.)W Watson (CF) 12 2.1.6.1 Botany 12 2.1.6.2 Chemical composition in CF 12 2.1.6.3 Pharmacological activities and the use of CF in traditional medicine 13 2.2 Application of organic materials in plant cultivation 13 2.3 The chemistry of phenolic compounds 16 2.3.1 Phenolic acid 17 2.3.2 Flavonoids 17 2.3.3 Tannins 20 2.4 Quantification of phenolics 20 2.4.1 Spectrophotometric assays 20 2.4.2 High performance liquid chromatogaraphy (HPLC) 21 2.5 Antioxidants 22 2.5.1 What is an antioxidant? 22 2.5.2 Basics of antioxidant characterization 23 2.5.3 Reactive oxygen species 23 2.5.4 Antioxidant assay 24 2.5.5 Synthetic phenolic and natural antioxidants 24 2.6 Introduction of cancers 25 2.6.1 Cancer 25 2.6.2 Introduction of human tumor cell lines 26 2.7 Introduction of apoptosis 27 2.7.1 What is apoptosis? 27 2.7.2 Bcl-2 family protein in apoptosis 28 2.7.3 Caspases in apoptosis 29 2.7.4 Effects of antioxidants on cancer 29 Chapter Research Methodology 32 3.1 The study setup 32 3.1.1 Study 32 3.1.2 Study 32 3.1.3 Study 32 3.1.4 Study 32 3.1.5 Study 32 3.2 Plant cultivation of Agrimonia pilosa Ledeb 33 3.3 Preparation of samples 35 3.4 Determination of total phenolic and flavonoid contents 36 Qin, C., Li, Y., Niu, W., Ding, Y., Zhang, R., and Shang, X (2010) Analysis and characterisation of anthocyanins in mulberry fruit CzechJournal of Food Science 28: 117-126 Ray, P., Huang, B., and Tsuji, Y (2012) Reactive oxygen species (ROS) homeostasis and redox regulation in cellular signaling Cell signal 24(5): 981990 Print Razali, N., Razab, R., Junit, S.M., and Aziz, A.A (2008) Radical scavenging and reducing properties of extracts of cashew shoots (Anacardium occidentale) Food Chemistry 111(1): 38-44 Re, R., Pellegrini, N., Proteggente, A., Pannala, A., Yang, M., and RiceEvans, C (1999) Antioxidant activity applying an improved ABTS radical cation decolorization assay Free Radical Biology and Medicine 26(9-10): 1231-1237 Reinmuth, N., Stumpf, P., Stumpf, A., Muley, T., Kongbinger, S., Hoffmann, H., and Thomas, M (2014) Characteristics of lung cancer after a previous malignacy Respiratory Medicine 108(6): 910-917 Robbins, R.J (2003) Phenolic acids in foods: An overview of analytical methodology Journal of Agricultural and Food Chemistry 51: 2866-2887 Romani, A., Campo, M., and Pinelli, P (2012) HPLC/DAD/ESI-MS analyses and anti-radical activity of hydrolyzable tannins from different vegetal species Food Chemistry 130: 214-221 Rostagno, M.A., Palma, M., and Barroso, C.G (2005) Solid-phase extraction of soy isoflavones Journal of Chromatography A 1076: 110-117 Santos, L.C.D., Sannomiya, M., Piacente, S., Pizza, C., Sano, P.T., and Vilegas, W (2004) Chemical profile of the polar extract of Paepalanthus microphyllus (Guill) Kunth (Eriocaulaceae) Brazilian Journal of Pharmaceutical Science40(3): 432-436 Salla, S., Sunkara, R., Ogutu, S., Walker, L.T., and Verghese, M (2015) Antioxidant activity of papaya seed extracts against H2O2 induced oxidative stress in HepG2 cells LWT-Food Science and Technology 66: 293-297 Salmani, J.M.M., Zhang, Z.P., Jacob, J.A., and Chen, B.A (2017) Apigenin’s anticancer properties and molecular mechanisms of action: Recent advances and future prospectives Chinese Journal of Natural Medicines 15(5): 0321-0329 Sarveswaran, S., Marati, R.V., and Maruthaiveeran, P.B (2006) Effects of Terminalia arjuna bark extract on apoptosis of human hepatoma cell line HepG2 World Journal of Gastroenterology 12: 1018-1024 Schinella, G.R., Tournier, H.A., Prieto, J.M., Mordujvich, De Buschiazzo, P., and Rios, J.L (2002) Antioxidant activity of antiinflammatory plant extracts Life Sciences 70: 1023-1033 Scordino, M., Di Muaro, A., Passerini, A., and Maccarone, E (2004) Adsorption of flavonoids on resins: cyanidin 3-glucoside Journal of Agricultural and Food Chemistry 52: 1965-1972 Seidel, C., Schnekenburger, M., Dicato, M., and Diederich, M (2014) Antiproferation and proapoptotic activities of 4-hydroxybenzoic acid-based inhibitors of histone deacetylases Cancer letter 343(1): 134-146 Shalby, A.S., and Razin A.M (1992) Dense cultivation and fertilization for higher yield of thyme (Thymus vulgaris L.) Journal of Agronomy and Crop Science 168: 243-248 Sharma, P.R., Mondhe, D.M., Muthiah, S., Pal, H.C., Shahi, A.K., Saxena, A.K., and Qazi, G.Z (2009) Anticancer activity of an essential oil from Cymbopogon flexuosus Chemico-Biological Interactions 179: 160-168 Singleton, V.L., Orthofer, R., and Lamuela-Raventos, R.M (1999) Analysis of total phenols and other oxidation substrates and antioxidants by means of folin-ciocaltue reagent Methods in Enzymology 299: 152-178 Shin, W.J., Lee, K.H., Park, M.H., and Seong, B.L (2010) Broadspectrum antiviral effect of Agrimonia pilosa extract on influenza viruses Microbiology and Immunology 54: 11-19 Shyu, M.H., Kao, T.C., and Yen, G.C (2008) Hsian-tsao (Mesona procumbens Heml.) prevents against rat liver fibrosis incluced by CCl via inhibition of hepatic stellate cells activation Food and Chemical Toxicology 46(12): 3707-3713 Shi, Y (2002) Mechanisms of caspase activation an inhibition during apoptosis Molecular Cell 9: 459-470 Shi-wei, Z., and Fu-zhen H (1991) The nitrogen uptake efficiency from 15N labeled chemical fertilizer in the presence of earthworm manure (cast) In Advances in management and conservation of soil fauna, ed G.K Veeresh, D Rajgopal, and C.A Viraktemath, 539-42 New Delhi: IBH Sharma, A.K (2003) Biofertilizers for Sustainable Agriculture Updesh Purohit for Agrobios Jodhpur 41-46 Siddhuraju, P (2007) Antioxidant activity of polyphenolic compounds extracted from defatted raw and dry heated Tamarindus indica seed coat LWT-Food Science and Technology 40(6): 982-990 Siddhuraju, P., and Becker, K (2003) Antioxidant properties of various extracts of total phenolic constituents from three different agroclimatic origins of drumsticl tree (Moringa oleifera Lam.) leaves Journal of Agriculture and Food Chemistry 51: 2144-2155 Silva, E.M., Pompeu, D.R., Larondelle, Y., and Rogez, H (2007) Optimisation of the adsorption of polyphenols from Inga edulis leaves on macroporous resins using an experimental design methodology Separation and Purification Technology 53: 274-280 Sinha, R.K., Herat, S., Valani, D., and Chuanhan K (2009) Vermiculture and sustainable agriculture American-Eur-asian Journal of Agricultural and Environmental Science, IDOSI Publication: 1-55 Sikwese, F.E., and Duodu, K.G (2007) Antioxidant effect of a crude phenolic extract from sorghum bran in sunflower oil in the presence of ferric ions Food Chemistry 104: 324-331 Singh, T., Sinha, N., and Singh, A (2013) Biochemical and histopathological effects on liver due to acute oral toxicity of aqueous leaf extract of Eclipta alba on female Swiss albino mice Indian Journal of Pharmacology 45(1): 61-65 Sperandio, S., De Belle, I., and Bredesen, D.E (2000) An alternative, nonapoptotic form of programmed cell death Proceedings of the National Academy of Science 97: 14276-14381 Stalikas, C.D (2007) Review: Extraction, Separation, and detettion methods for phenolic acids and flavonoids Journal of Separation Science 30: 3268-3295 DOI:10.1002/jssc.200700261 Stutte, G.W (2006) Process and product recirculation hydroponics and bioactive compounds in controlled environment Horticultural Science 41: 526-530 Subler, S., Edwards, C., and Metzger J (1998) Comparing vermicomposts and composts Biocycle 39: 63-66 Suresh, K.D., Sneh, G., Krishn, K.K., and Mool C.M (2004) Microbial biomass carbon and microbial activities of soils receiving chemical fertilizers and organic amendments Archives of Agronomy and Soil Science 50: 641647 Swick, R.W., Baumann, C.A., Miller, W.L.J., Rnmsfeld, H.W.J (1951) Tocopherol in tumor tissues and effects of tocopherol on the development of liver tumors Cancer Research 11(12): 948-953 Print Tahami, Z.S.M.K., Moghaddam, P.R., and Jahan, M (2011) Effects of organic and chemical fertilizers on yield and percentage of essential oils content of the medicinal plant basil The Journal of Agricultural Econogy 2(1): 63-74 Tait, S.W., and Green, D.R (2010) Mitochondria and cell death: Outer membrane permeabilization and beyond Nature Reviews Molecular Cell Biology 11: 621-632 Takasaki, M., Tokuda, H., Nishino, H., and Konoshima, T (1999) Cancer chemopreventive agents (antitumor-promoters) from Ajuga decumbens Journal of Natural Products 62(7): 972-975 Thornberry, N.A., and Lazebnik, Y (1998) Caspases: enemies within Science 281 : 1312-1316 Tomati, U., Galli, E., Grappelli, A., and Di Lena G (1990) Effects of Earthworm Casts on Protein Synthesis in Radish (Raphanus sativum) and Lettuce (Lactuca sativa) Seedlings Biology and Fertility of Soils 9: 288-289 Doi: org/10.1007/BF00634102 Tsai, T.H., Tasi, P.J., and Ho, S.C (2005) Antioxidant and antiinflammatory activities of several commonly used spices Journal of Food Science 70(1): 93-97 Tsai, M.L., Lin, C.C., and Yang, C.H (2011) Antimicrobial, antioxidant, and anti-inflammatory activities of essential oils from five selected herbs Bioscience, Biotechnology, and Biochemistry 75: 1977-1983 Uttara, B., Singh, A.V., Zamboni, P.A., andMahajan, R.T (2009) Oxidative stress and neurodegenerative diseases: A review of upstream and downstream antioxidant therapeutic options Current Neuropharmacology 7: 65-74 Velioglu, Y.S., Mazza, G., and Oomah, B.D (1998) Antioxidant activity and total phenolics in selected fruits, vegetables, and grain products Journal of Agricultural and Food Chemistry 46: 4113-4117 Venderjagt, T.J., Ghattas, R., Vanderjagt, D.J., Crossey, M., and Glew, R.H (2002) Comparison of the total antioxidant content of 30 widely used medicinal plants of New Mexico Life Science 70: 1035-1040 Wang, X (2001) The expanding role of mitochondria in apoptosis Genes and Developments 15: 2922-2933 Wagner, H., Geyer, B., Kiso, Y., Hikino, H., and Rao, G.S (1986) Coumestans as the main active principles of live drugs Eclipta alba and wedelia celendulacea Planta Medica 53: 370-374 Walkley Black, A., and Black, L.A (1934) An examination of the Dgtiarff method for determining soil organic matter, and a proposed modification of the chromic acid titration method Soil Science 37: 28-29 Warncke, D.D (1986) Analyzing greenhouse growth media by the saturation extraction method Horticultural Science 21: 223-225 Wintermans, J.F., and De Mots (1965) Spectrophotometric characteristic of chlorophylls a and b and their pheophytins in ethanol Biochimica et Biophysica Acta 109(2): 448-453 Wurstle, M.L., Laussmann, M.A., and Rehm, M (2012) The central role of initiator caspase-9 in apoptosis signal transduction and the regulation of its activation and activity on the apoptosome Experimental Cell Research 318: 1213-1220 Xia, G.H., Chen, B.A., Shao, Z.H Lu, H.X., Konstanze, D.,and Hartmut, D (2007) Mechanism of 2-methoxyestradiol-induced apoptosis in myelodysplastic syndrome MUTZ-1 cell line Journal of Experimental hematology 15: 296-301 Xiao, D.J., Zhu, G.C., Wang, Y.P., and Hu, Y.L (2008) Inhibiting effect of Hypericum japonicum Thunb on proliferation of human nasopharyngeal carcinoma line CNE-2 in vitro Mod Oncol 16: 15-16 Xie, X., Wang, Y.L., Zhu, D.H., Feng, X.L.,and Huang, L.F (2013) Analysis of the volatile chemical constituents in Agrimonia pilosa Ledeb by gas chromatography–mass spectrometry with Kovats retention indices Fenxi Kexue Xuebao 29: 651-654 Xu, X., Qi, X., Wang, W.,and Chen., G (2005) Separation and determination of flavonoids in Agrimonia pilosa Ledeb by capillary electrophoresis with electrochemical detection Journal of Separation Science28: 647-652 Xu, B.J, and Chang, S.K.C (2007) A Comparative Study on Phenolic profiles and Antioxidant Activities of Legumes as Affected by Extraction Solvents Journal of Food Science72(2): 159-166 Yahara, S., Ding, N., Nohara, T., Masuda, K., and Ageta, H (1997) Taraxastane glycosides from Eclipta alba Phytochemistry 44: 131-135 Yamaki, M., Kashihara, M., Ishiguro, K., and Takagi, S (1989) Antimicrobial principles of Xian he cao (Agrimonia pilosa) Planta Media 55: 169-170 Yang, J., Guo, J., and Yuan, J (2008) In vitro antioxidant properties of rutin TWT-Food Science and Technology 41(6): 1060-1066 Yanishlieva-Maslarova, N.V (2001) Inhibiting oxidation In J Pokorny, N Yanishlieva, & M.H Gordon (Eds.), Antioxidants in food: Practical applications (pp 22-70) Cambridge: CRC Press, Woodhead Publishing Limited Yedjou, C.G., Moore, P., and Tchounwou, P.B (2006) Dose and Time Dependent Response of Human Leukemia (HL-60) Cells to Arsenic Trioxide Treatment International Journal of Enviromental Research and Public Health 3(2): 136-140 Yen, G.C., Hung, Y.L., and Hsieh, C.L (2000) Protective effect of extracts of Mesona procumbens Hemsl on DNA damage in human lymphocytes exposed to hydrogen peroxide and UV irradiation Food and Chemical Toxicology 38: 747-754 Yen, G.C., Hung, Y.L., and Chen, Y.J (2003) Antioxidant Properties of Hsian-tsao (Mesona procumbens Hemsl.) Oriental Foods and Herbs 14, 202214 Yen, G.C., Duh, P.D., and Hung, Y.L (2001) Contributions of major components to the antimutagenic effect of Hsian-tsao (Mesona procumbens Hemsl.) Journal of Agricultural and Food Chemistry 49(10): 5000-5004 Zarena, A.S., and Udaya Sankar, K (2011) Phenolics acids, flavonoid profile and antioxidant activity in mangosteen (Garcina mangostana L.) pericarp Journal of Food Biochemistry 36: 627-633 Zhang, M., Chen, Y.Y., Di, X.H., and Liu, M (1997) Isolation and identification of Ecliptasaponin D from Eclipa alba (L.) Hassk Acta Pharmaceutica Sinica 32: 633-634 Zhang, J.S., and Guo, Q.M (2001) Studies on the chemical constituents of Eclipta prostrate (L) Acta Pharmaceutica Sinica 36: 34-37 Zhu, L., Tan, J., Wang, B., He, R., Liu, Y., and Zheng, C (2009) Antioxidant activities of aqueous extract from Agrimonia pilosa Ledeb and its fractions Chemistry and Biodiversity 6: 1716-1726 Appendices Appendix Mass spectrum of compounds SA3 and SA4 Inten.(x1,000,000) 259.15 2.5 215.10 2.0 1.5 245.15 338.45 0.5 0.0 200 383.05 303.50 1.0 289.10 250 417.30 361.20 300 350 400 455.45 527.45 622.85 543.95 450 500 671.60 550 600 650 m/z SA3-ESI+ Inten.(x10,000,000) 1.00 0.75 0.50 383 464 401 423 0.25 0.00 200 237 250 282 311 300 350 527 543 450 338 400 SA4-ESI+ 450 500 550 6509 m/z Bio-Sketch of Author Full name: Le Quang Ung Gender: Male Nationality: Vietnamese Education background Education Major Date Institution Ph.D Master Herbal medicine and natural products Crop Sciences Bachelor Crop Sciences 2015 – National Pingtung University of 2019 Science and Technology, Taiwan 2011– 2013 2006– 2011 Thai Nguyen University, Vietnam Thai Nguyen University, Vietnam Published Papers Le, Q.U., Lay, H.L., and Wu, M.C (2017) Antioxidant activities and HepG2 cells growth inhibitory capacity of whole plant ethanol extracts (Eclipta alba Hassk and Mesona procumbens Hemsl) Journal of Food Biochemistry, Doi: 10.1111/jfbc.12454 42(2) (SCI paper) Le, Q.U., Lay, H.L., and Wu, M.C (2018) The isolation, structural characterization and anti-cancer activity from the aerial parts of Cymbopogon flexuosus Journal of Food Biochemistry Doi: 10.1111/jfbc.12718 (SCI paper) Le, Q.U., Lay, H.L., Wu, M.C., Nguyen, D.L., and Nguyen, T.H.H (2018) Phytoconstituents and Biological Activities of Panax vietnamensis (Vietnamese Ginseng): A Precious Ginseng and Call for Further Research-A systematic review Natural product communications 13(10), 1381-1384 (SCIE paper) Le, Q.U., Lay, H.L., and Wu, M.C (2018) Comparative study of Anti-oxidant and alpha amylase Inhinitory Activity of Six herbs used in traditional medicine International Journal of Scientific and Engineering Research, 9(3), 510-529 Le, Q.U., Lay, H.L., and Wu, M.C (2018) Effects of Vermicompost and Hog Manure on Growth, Yield and Antioxidant Activities of Agrimonia Pilosa Ledeb as a medicine plant Biological forum- An International Journal, 10(1), 56-63 (Web of Science Platform) Le, Q.U., Joshi, R.K., Lay, H.L., and Wu, M.C (2018) Agrimonia pilosa Ledeb: Phytochemistry, Ethnopharmacology, Pharmacology of an important traditional herbal medicine Journal of Pharmacognosy and Phytochemistry, 7(4), 3202-3211 Le, Q.U., Lay, H.L., and Wu, M.C (2018) Antioxidant activities and A549 growth inhibitory capacity from various extracts of Agrimonia pilosa Ledeb Journal of Ayurvedic and herbal medicine.4(3), 114-119 Le, Q.U., Lay, H.L., Wu, M.C., and Nguyen, T.H.H (2018) Natural plant colorants widely used widely in Vietnam traditional food culture Journal of Food, nutrition and Agriculture 1(1), 40-46 Le, Q.U., Lay, H.L., Wu, M.C., Nguyen, D.L., and Nguyen, T.H.H (2018) Hovenia dulcis Thunb revisited-A mini critical review and call for further research to insightfully elucidate Journal of Pharmacognosy and Phytochemistry 7(6), 1137-1141 10 Le, Q.U., Lay, H.L., and Wu, M.C (2019) Antioxidant potential and HepG2 and A549 cell growth inhibitory capacity of Hypericum japonicum Thunb and Scoparia dulcis L Journal of medicinal plants studies.Accepted 11 Le, Q.U., Lay, H.L., and Wu, M.C (2019) Medicinal botanicals in the osteoarthritis treatment and management in traditional Vietnamese medicine International Journal of Botany Studies 4(3), 59-62 12 Le, Q.U., Lay, H.L., and Wu, M.C (2018) Phytoconstituents and pharmacological activities of Silybum marianum (Milk Thistle): A critical review American Journal of Essential Oils and Natural Products 6(4):41-47 13 Le, Q.U., Lay, H.L., and Wu, M.C (2019) Herbs for the Management of Diabetes Mellitus in Traditional Vietnamese Medicine Journal of Applied Biopharmaceutics and Pharmacokinetics 7, 1-6 ... unilocular, about 2-3 mm long, ovules many, styles 3, free from the base, erect, about 1.2 mm long Fruit capsule septicidal, ovoidsubglobose, about 4-4.5 x 2.5-3 mm long, apex obtuse, longitudinally vittate,... 4-lobed, lateral lobes distinct, longer than middle lobes, or upper lip subentire Anterior stamens longer than basally hirsute posterior pair Nutlets black, oblong Flowering and fruiting are from... pedicel about 4-7 mm long, bracts linear lanceolate, foliaceous, about 2-2.5 mm long, caduceus, sepals 5, unequal, quincuncial, outer 2, lanceolate-ovate, inner 3, oblanceolate-oblong, chartaceous,