initial characterization of crude extracts from phyllanthus amarus

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for the TLC separation of most of the plant extracts and plant drug extracts (Barbetti et al., 1986; Houghton et al.1996; Wagner and Bladt, 1996). When using a complex mixture such as a biological sample, the concentration of the solute of interest is often not known. Trial and error is needed when applying samples to a TLC plate in order to determine the optimal concentration for spotting, which is usually governed by the method of detection employed (Fried and Sherma, 1994). 22
CHAPTER 3 MATERIALS AND METHODS 3.1. Collection of Plant Material Phyllanthus amarus Schum. and Thonn. and Quassia amara L. were collected in Suriname. Since there are no cultivated fields of these plants in Suriname, the plant material had to be collected from different geographical locations. The plants were (haphazardly) sampled from areas where they grow naturally. The environmental conditions under which the plants grow (pasture land, woods), the soil composition, and pH of the soil were recorded (Appendix A). Plants of P. amarus were collected (June; July 2001) in the city (Paramaribo- North (PN) and Paramaribo-South (PZ)) as well in the country (Saramacca-Damboentong (PD)). Q. amara (wood) was collected (July 2001) in the countryside (Saramacca- Dirkshoop). In August 2001 the dried plant species were shipped to Baton Rouge. 3.2. Sample Preparation and Extraction P. amarus material was air dried under shade for two weeks and Q. amara (wood) was oven dried one week at 40°C. The dried plant material was ground (October; November 2001) with a Wiley Mill grinder (Standard Model No. 3). P. amarus was ground to 2 mm or smaller particle size. The wood of Q. amara was ground to 6 mm or smaller particle size. The ground plant species were stored in a refrigerator at 4°C. Crude Extracts To investigate if there is a difference in the number of compounds recovered in the samples of P. amarus from the different geographical locations (PD, PN, PZ), each sample was extracted separately. From each location 1 gram of plant material was extracted (in May 2002) with 10 ml (1:10, w/v) of either solvent A, 50% methanol in deionized and distilled (D.D) water; B, 99% methanol; C, 50% methanol in chloroform. 23
Page 1 and 2: INITIAL CHARACTERIZATION OF CRUDE E
Page 3 and 4: suggestions to the project and manu
Page 5 and 6: ABSTRACT The extracts of many plant
Page 7 and 8: CHAPTER 1. INTRODUCTION According t
Page 9 and 10: 2.1. Background CHAPTER 2. LITERATU
Page 11 and 12: In drug discovery, the major second
Page 13 and 14: greenish, often with glands. The fr
Page 15 and 16: wood is soft and used for boxes, ki
Page 17 and 18: used for colic (Wessels Boer, 1976;
Page 19 and 20: phytochemical compounds as possible
Page 21 and 22: and fuel. In Puerto Rico the wood o
Page 23 and 24: extracts from the dry quassia wood
Page 25 and 26: Quassin has important pharmaceutica
Page 27: alumina, cellulose powder, polyamid
Page 31 and 32: 1. Concentration 2. LPE 3× with he
Page 33 and 34: II. At UV-365 nm the spots were cir
Page 35 and 36: Twenty-five milligrams of crude pow
Page 37 and 38: still not completely separated. Mob
Page 39 and 40: indication that fraction 3 is less
Page 41 and 42: concentration of them is too low to
Page 43 and 44: and then visualizing the spots at U
Page 45 and 46: other impure non polar fractions/co
Page 47 and 48: to three (Q. amara) fraction(s)/com
Page 49 and 50: the extraction solvent 50% methanol
Page 51 and 52: HCl, the filtrate was then basified
Page 53 and 54: than TLC in analyzing the specific
Page 55 and 56: Dayan, F.E., Watson, S.B., Galindo,
Page 57 and 58: Mbwambo, Z.H., Luyengi, L., Kinghor
Page 59 and 60: Unander, D.W., Webster, G.L. and Bl
Page 61 and 62: APPENDIX B. EXTRACTION LAYOUT Table
Page 63 and 64: APPENDIX C. THIN LAYER CHROMATOGRAP
Page 65 and 66: Figure C.5 Fractions from crude P.
Page 67 and 68: Figure C.9 Fractions from crude P.
Page 69 and 70: 9:1 95:5 98:2 Figure C.13 Fractions
Page 71 and 72: APPENDIX D. RF-VALUES OF FRACTIONS
Page 73 and 74: Table D.3 Rf-values of the fraction
Page 75 and 76: Table D.6 Rf-value of fractions in

initial characterization of crude extracts from phyllanthus amarus

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