Basic Phytochemical Techniques “Laboratory Mana 5 Co Sehen a Saxhlet Extraction Thin Layer Chromatography — Supereriical Fluid Chromatography Photos: Puja LN, Sharma Gautam sot, Reseaeh OMicer Depnvtieoat of Plant Resources Noiural Freduct Reseatvlt Laboratory, ThapathaltBasic Phytochemical Techniques Laboratory Manual By L.N. Sharma Gautam Asst. Research Officer Natural product research Laboratory, ‘Thapaths | cs | | Z soviet. moto Copy Rights Reserved By Author and Department of Plant Resources, Natural Product Research Laboratory, Thapathali (NPRL), NepalPREFACE Since this manual is a compilation of the basic works necessary for Phytochemical Techniques, it will meet the needs and requirements of those graduates like Chemist, Pharmacist, Botanist & Bio-chemist who are working at various plants research laboretoriee as well es for pharmacy and chemistry student studying in various levels at diferent univenitiee and institutions, This manual is preliminary 8 premeusite for phytochemistry. Te has heen mine endeavor to present the basic procedure for Phytochemistry Research so as to make the laboratory work as methodical and efficent 3 posible acknowledge matters taken from other books, articles and intemets. 1 sincerely hope that this manual wil be of great help to the respective personalities, I wwelzome suggestions from the users to improve the quality ofthis volume which wll be incorporated in its next edition, I am much indepted to Prof. M. B. Gewali, R. N. Jha, B. M. ‘Tamrakar, and R. Adhihari for their valuable hours for correcting, writing and providing suggestions, My sincere thanks go to my family members ‘or encouragement and aspirations ohh, Nath, Sharma Gautem NPRI January, 2007 Phone no. ol- 4268247(Of)‘ACRONYMS, ams Grams mp. 2 Mebing pon = Maer mg 2 Miligam mi > silter nm Nasometer Dm “Maximam Wavelength Vow Maximum Frequency x 2 Ratio of Front The Thin Layer Chromatography ow Ula Violet EOAC tiylaceate MeOH Methanol EOH Edasol CHCl, Chloroform Pot Preiitation NusSO Sodium sulphate ec, Carbon evaelorde cH.cooH ‘etc acid FeSO, Feous sulphate Rec Fens chloride BuO ‘Buianol Mae Insolble residue cs: Carbonisulphide NaiCO, Sodium carbonate Sec AAmimony chionde Hs “Hydrogen sulphide caso. Copper sulphate Nall Sodium hyaroxide HO Sulpburis acid NiLOH “Ammonium hydroxide Cl Hydrochloric acidCONTENTS 1. Ineroduetion 1.1 Medicinal plan research methodology 1.2 Extraction technique 13 General methods of Isolation 14 Phytocherical screening 2. Separation Techniques 2.1 Thin layer chromatography 22 Column chromatography 3. olation Techniques 3.1 Taxol from Taxus baccata 3.2 Wededolactone from Eclipra alba 3.3 Valeranone & Spirojetamol fiom Nardostacysjatamanst 3.4 Kutkoside and Pirosie I fom Pierorhica burrow 3.5 Flavonoids from dry Alinom cepa 3.6 Uanic acid from usnea species 3.7 Bergenin from Bergenia ciliate 3.8 Stigmasterol from Gieine max 3.9 Nicotine from Nicotiana tobaccum 3.10 Lecithin from Ege Yolk 3:11 Lycopene from Lycopersicum esculentum 3112 Lactose and easein ftom milk 3113 Essential ol fom Eucalypru leaves 3.14 Dimethoxy aromadendin from Podacarpus ner 4 Appendices 5, References