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Final Report Final Report CHEMICAL STUDIES AND BIOLOGICAL INVESTIGATIONS OF VIETNAMESE PLANTS AND HERBAL MEDICINES FROM THE EUPHORBIACEAE FAMILY TO DEVELOP HIGH-VALUE HEALTHCARE MATERIALS, AND THEIR QUALITY CONTROL (BL/03/V21) Period December 2009 – September 2012 Partners: Belgium : (coordinator and divers partners) Flemish partner : Prof. Yvan Vander Heyden (project leader) Vrije Universiteit Brussel (VUB) – FABI Laarbeeklaan 103, B-1090 Brussels, Belgium [email protected] Walloon partner: Professor Joëlle Quetin-Leclercq Université Catholique de Louvain (UCL) Analytical chemistry, drug analysis and pharmacognosy unit (CHAM) Avenue E. Mounier 72, B-1200 Brussels, Belgium [email protected] International partners: Vietnamese partners: Professor Dr. Chau Van Minh (project leader) Vietnam Academy of Science and Technology VAST 18 Hoang Quoc Viet Road, Nghia Do - Cau Giay - Hanoi Tel. 84-4-8363375 Fax 84-4-7568171 [email protected] [email protected] Dr. Nam Nguyen Hoai Vietnam Academy of Science and Technology (VAST) Institute of Marine Biochemistry (IMBC) 18 Hoang Quoc Viet - Cau Giay - Hanoi Tel: +84912260146 [email protected] CONTENT Content General Introduction Final Report Conclusions Results Sheet Annexes VAST-IMBC Annexes UCL-CHAM Annexes VUB-FABI GENERAL INTRODUCTION General Introduction 1 Scientific background Natural products have served mankind as a source of medicine since or even before the recorded history existed. Vietnam, with its vast geographical diversity, boasts diverse botanical resources and possesses a strong ethno-botanical tradition. About 3200 medicinal plants grow in Vietnam with a third regarded as medicinally effective and the remaining two third as dubious to mildly effective. Those resources have a great potential to be developed as high-value healthcare products (food supplements). Nowadays, natural products are also gaining more interest in Western countries. Quality control is an important issue during development of such products to ensure the identity, safety and quality of the natural and derived products. In this regard we have made efforts to survey useful medicinal plants, and other bio-resources from Vietnam and to evaluate their potential economic value using state-of-the-art chemical and molecular biological techniques. Natural compounds of vegetable origin with interesting biological activities were examined, in order to be possibly utilized as new drugs or in the para-pharmaceutical field, e.g. as non- nutritive and alternative sweeteners, nutraceuticals or ecological insecticides. Only a limited number of natural products was focused on and considered as case studies. The search for bio-active components from natural materials as a source of lead compounds in drug development is a major endeavor in natural products chemical research. Some species belonging to the Euphorbiaceae family, for example Phyllanthus and Mallotus species , are very important in the Vietnamese traditional medicine and have diverse activities. In a herb and its extracts, there are hundreds of unknown components, many of them are in low amounts and usually variability exists within the same herbal material. Moreover, the chemical components in the herbal medicinal products may vary depending on the harvest season, plant origins, drying processes and other factors. Therefore assaying only some isolated compounds and/or pharmacologically active constituents hardly represents the complex herbal extracts and is not reliable enough for the quality control of these herbs. The actual situation is that many herbs are used in traditional medicine, but that their active principles (or derived compounds) are not commercialized because either they were not identified yet or their efficacy and safety is insufficiently examined and documented yet or because a synergy or additivity of effects exists between several compounds or classes of compounds, needing the use of more crude extracts (polytherapy). The project focused on the chemical studies and biological investigations of Vietnamese plants and herbal medicines from the Euphorbiaceae family to develop healthcare materials (e.g. food supplements), and on their quality control. 2 Overall objectives A number of natural products from the Euphorbiaceae family, i.e. Mallotus and Phyllanthus species, were focused on and considered as case studies. Therefore, the main aims of this project, entitled “Chemical and biological investigations of Vietnamese herbal medicines from the Euphorbiaceae family to develop high-value healthcare materials and their quality control”, were: 1) To find natural compounds or extracts from Euphorbiaceae plants used in Vietnam as herbal medicines with interesting biological activities to contribute as new material for the pharmaceutical or food industries, e.g. as food material, functional food, nutraceuticals, … 2) To survey Vietnamese medicinal plants from the Euphorbiaceae family used in traditional medicine in Vietnam. The study focused mainly on Phyllanthus , but also on Mallotus species, with a special interest in endemic species. 3) To study the chemical composition and investigate biological activities of selected medicinal plants using chemical and molecular biological techniques. 4) To produce materials (food supplements) and/or to identify lead molecules with a potential interest for pharmaceutical or food industries. 5) To apply analytical aspects for quality control of plant materials or products from plants: chromatographic fingerprints of crude extracts for identification and quality control purposes, and quantitative determination of specific active compounds. 3 Specific tasks 3.1 VAST-IMBC - To survey and collect potentially medicinal plants of the Phyllanthus species. This occasionally also will be done for Mallotus species. - Preparation of crude and/or purified extracts of selected plant samples and primary screening extracts. Including defined and modified methodology to produce Phyllanthus and Mallotus extracts. - Isolation and structure determination of compounds from the selected plants. - Chromatographic fingerprint with isolated compounds as markers. 3.2 UCL-CHAM - Evaluation of some biological activities of Mallotus and Phyllanthus species. - Isolation and identification of active constituents. - Analysis of extracts by HPLC-MS. 3.3 VUB-FABI - Define a methodology to develop HPLC fingerprints. - Develop HPLC-DAD fingerprints of a set of Mallotus and/or Phyllanthus samples, herbs commonly used in Vietnamese traditional medicine, and link the HPLC fingerprints to measured activities, such as antioxidant, antimicrobial and cytotoxic activity. For this purpose, prediction models will be build using linear multivariate calibration techniques and it was aspired to indicate and identify the peaks and their underlying compounds that are potentially responsible for the measured activity. - Evaluate pressure-assisted capillary electrochromatography (pCEC-UV) as a complementary technique to HPLC for the indication of possible antioxidant compounds in Mallotus samples from different species. - Discriminate between Mallotus and Phyllanthus species (two classes) and their sub-species (six classes). - Develop UFLC-DAD fingerprints of a set of Mallotus and/or Phyllanthus samples on a fused- core column, and link the HPLC fingerprints to measured activities, such as antioxidant, antimicrobial and cytotoxic activity. For this purpose, prediction models will be build using linear multivariate calibration techniques and it was aspired to indicate and identify the peaks and their underlying compounds that are potentially responsible for the measured activity. FINAL REPORT Bilateral scientific cooperation W&T-cooperation with Vietnam BL/03/V21 Final Report Period December 2009 – September 2012 11 1. Administrative information 1.1. Project nr. BL/03/V21 1.2. End date of the Project September 30th 2012 1.3. Partner country Vietnam (Prof. Chau van Minh) 1.4. Flemish promotor - Name Prof. Yvan Vander Heyden 1.5.Participating Waloon universities Université Catholique de Louvain (Prof. Joelle Leclercq-Quetin) 2. Summary of the Research Results A number of natural products from the Euphorbiaceae family, i.e. Mallotus and Phyllantus species, were focused on and considered as case studies. Therefore, the main aims of this project, entitled “Chemical and biological investigations of Vietnamese herbal medicines from the Euphorbiaceae family to develop high-value healthcare materials and their quality control”, were: 1) To find natural compounds or extracts from Euphorbiaceae plants used in Vietnam as herbal medicine with interesting biological activities to contribute as new material for the pharmaceutical or food industries, e.g. as food material, functional food, nutraceuticals, … and 2) To survey Vietnamese medicinal plants from the Euphorbiaceae family used in traditional medicine in Vietnam. The study will focus mainly on Phyllanthus, but also on Mallotus species, with a special interest in endemic species. 3) To study the chemical composition and investigate biological activities of selected medicinal plants using chemical and molecular biological techniques. 4) To produce materials (food supplements) and/or to identify lead molecules with a potential interest for pharmaceutical or food industries. 5) To apply analytical aspects for quality control of plant materials or products from plants: chromatography fingerprints of crude extracts for identification and quality control purposes, and quantitative determination of specific active compounds. 2.1.
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