Laboratory Handbook for the Fractionation of Natural Extracts

Peter J. Houghton
and

Amala Raman
Pharmacognosy Research Laboratories, Department of Pharmacy, King's College London

CHAPMAN & HALL

London Weinheim New York Tokyo Melbourne Madras

Contents
1 Introduction 1.1 The need for this book - an active extract, what happens next? 1.2 Why is it important to find out the nature of the active components? 1.3 Green chemicals - the renewed interest in natural products as sources of bioactive compounds 1.4 What are we doing? 1.5 Bibliography 2 Important concepts 2.1 Introduction 2.2 Polarity ^ 2.3 Dilution and concentration factors 2.4 Synergy 2.5 Deterioration, decomposition and storage 2.6 Biological variation 2.7 Authentication of material 2.8 Dereplication 3 Methods for extraction and sample clean-up 3.1 Introduction 3.2 Factors to be considered in selecting an extraction method 3.3 General extraction methods 3.4 Extraction methods for specific phytochemical groups 3.5 Clean-up techniques to remove unwanted substances 3.6 Bibliography 4 Crude fractionation procedures 4.1 Introduction 4.2 Choice of fractionation method 4.3 Common fractionation methods 4.4 Bibliography 1 1 2 4 7 13 14 14 14 14 16 17 18 20 21 22 22 22 27 39 45 52 54 54 55 56 65

vi

Contents
66 66 66 71 81

5 Chromatographic fractionation procedures 5.1 Introduction 5.2 The principles of chromatography 5.3 Physico-chemical processes taking place in chromatography 5.4 Bibliography

6 Chromatographic procedures using columns and liquid-liquid systems 82 6.1 Column chromatography 82 6.2 Liquid-liquid partition schemes 90 6.3 Bibliography 93 7 Layer chromatography 7.1 Layer chromatography 7.2 Preparative thin-layer chromatography and paper chromatography 7.3 Bibliography 8 Analysis of crude extracts, fractions and isolated compounds 8.1 Introduction 8.2 Qualitative analysis of mixtures and isolates 8.3 Quantitative analytical methods 8.4 Biological assays 8.5 Bibliography 9 General methods 9.1 Basic techniques in handling extracts 9.2 Isolation of active compounds 9.3 Chromatographic procedures 9.4 Basic techniques in thin-layer chromatography 9.5 Bibliography 10 Practical exercises in chemical screening and fractionation of crude extracts from plant materials 10.1 Chemical screening 10.2 Exercises in extraction, clean-up, fractionation and chromatographic methods Appendix A Recipes for chemical test reagents Index 94 94 102 112 113 113 114 127 135 138 139 139 145 146 150 153

154 154 162 184 187