ISOLATION AND CHARACTERIZATION OF ALKALOIDS: STUDY OF DELPHINIUM CHITRALENSE AND RELATED MEDICINAL PLANTS BY Mr. SHUJAAT AHMAD DOCTOR OF PHILOSOPHY (PhD) IN CHEMISTRY DEPARTMENT OF CHEMISTRY UNIVERSITY OF MALAKAND 2016 ISOLATION AND CHARACTERIZATION OF ALKALOIDS: STUDY OF DELPHINIUM CHITRALENSE AND RELATED MEDICINAL PLANTS BY Mr. SHUJAAT AHMAD Thesis submitted to the Department of Chemistry, University of Malakand for the partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY (PhD) IN CHEMISTRY DEPARTMENT OF CHEMISTRY UNIVERSITY OF MALAKAND 2016 Declaration I declare that the thesis “ISOLATION AND CHARACTERIZATION OF ALKALOIDS: STUDY OF DELPHINIUM CHITRALENSE AND RELATED MEDICINAL PLANTS” is my original work and has never been presented for the award of any degree at any other University before and that all the information sources I have used and or quoted have been indicated and acknowledged by means of complete references. Shujaat Ahmad Certificate It is recommended that this thesis submitted by Mr. SHUJAAT AHMAD entitled “ISOLATION AND CHARACTERIZATION OF ALKALOIDS: STUDY OF DELPHINIUM CHITRALENSE AND RELATED MEDICINAL PLANTS” be accepted as fulfilling this part of the requirements for the degree of Doctor of Philosophy (PhD) in Chemistry. ______________________ _______________________ SUPERVISOR EXTERNAL EXAMINER Dr. Manzoor Ahmad Associate Professor ____________________ CHAIRMAN DEPARTMENT OF CHEMISTRY Dedicated To ALL PEACE AND KNOWLEDGE LOVING PEOPLE Contents ACKNOWLEDGMENTS .................................................................................................... i SUMMARY ........................................................................................................................ iii List of Table ....................................................................................................................... vii List of Figures ................................................................................................................... viii List of Scheme ..................................................................................................................... x List of abbreviations ........................................................................................................... xi Chapter-1 ............................................................................................................................. 1 1.1 Introduction .............................................................................................................. 1 1.2 Plants as drugs sources .............................................................................................. 2 1.3 Medicinal plants and Islamic literature ..................................................................... 3 1.4 Plants as traditional medicine and drugs discovery .................................................. 6 1.5 Alkaloids.................................................................................................................... 6 1.5.1 Diterpene alkaloids ......................................................................................... 7 1.5.2 C20 diterpene alkaloids .................................................................................... 7 1.5.3 C19 diterpene alkaloids .................................................................................... 9 1.5.4 C18 diterpene alkaloids .................................................................................. 11 1.6 Therapeutic potential of diterpene alkaloids ........................................................ 11 1.7 The Genus Delphinium ........................................................................................ 13 1.7.1 Systematic position .............................................................................................. 14 1.7.2 Delphinium chitralense ........................................................................................ 14 1.7.3 Delphinium denudatum Wall. .............................................................................. 15 1.7.4 Literature survey of genus Delphinium ............................................................... 16 Chapter-2 EXPERIMENTAL ..................................................................................... 20 2.1 General experimental conditions ............................................................................. 20 2.1.1 Physical constants ......................................................................................... 20 2.1.2 Spectroscopic techniques .............................................................................. 20 2.1.3 Column chromatography (CC) ..................................................................... 20 2.1.4 Solvents ......................................................................................................... 20 2.1.5 Detection of alkaloids on chromatographic plates ....................................... 21 2.2. Plant materials ......................................................................................................... 21 2.2.1 Plants materials pretreatment ........................................................................ 21 2.2.2 Extraction ...................................................................................................... 21 2.2.3 Fractionation ................................................................................................. 22 2.2.4 Isolation of alkaloids from D. chitralense .................................................... 24 2.2.5 Isolation of alkaloids from D. denudatum Wall. .......................................... 25 2.3 X- rays diffraction studies ....................................................................................... 26 2.4 Assay for enzymatic inhibition and IC50 value determination ................................ 26 2.5 Physical and spectroscopic data of new compounds from D. chitralense .............. 28 2.5.1 Chitralinine-A (1) ......................................................................................... 28 2.5.2 Chitralinine-B (2).......................................................................................... 28 2.5.3 Chitralinine-C (3).......................................................................................... 29 2.5.4 Chitraline-A (4) ............................................................................................ 29 2.5.5 Chitraline-B (5) ............................................................................................. 30 2.5.6 Chitraline-C (6) ............................................................................................. 31 2.5.7 Chitraline-D (7) ............................................................................................ 31 2.6 Physical and spectroscopic data of new compounds from D. denudatum Wall. .... 32 2.6.1 Compound (8) ............................................................................................... 32 2.6.2 Compound (9) ............................................................................................... 33 2.7 Physical and spectroscopic data of known compounds from D. Chitralense. ........ 33 2.7.1 Ajaconine (10) .............................................................................................. 33 2.7.2 Delectinine (11) ............................................................................................ 34 2.8 Physical and spectroscopic data of known compounds from D. denudatum Wall. 34 2.8.1 Dihydropentagynine (12) .............................................................................. 34 Chapter-3 RESULTS AND DISCUSSION ......................................................... 36 3.1 Present work ............................................................................................................ 36 3.2 New diterpenoids from Delphinium chitralense ..................................................... 36 3.2.1 Chitralinine-A (1) ......................................................................................... 36 3.2.2 Chitralinine-B (2).......................................................................................... 43 3.2.3 Chitralinine-C (3).......................................................................................... 47 3.2.4 Chitraline-A (4) ............................................................................................ 51 3.2.5 Chitraline-B (5) ............................................................................................. 55 3.2.6 Chitraline-C (6) ............................................................................................. 59 3.2.7 Chitraline-D (7) ............................................................................................ 63 3.3 New diterpenoids from D. denudatum wall. ........................................................... 67 3.3.1 Compound (8) ............................................................................................... 67 3.3.2 Compound (9) ................................................................................................ 71 3.4 Known diterpenoids from D. chitralense ................................................................ 74 3.4.1 Ajaconine (10) .............................................................................................. 74 3.4.2 Delectinine (11) ............................................................................................ 77 3.5 Known diterpenoids from D. denudatum Wall ......................................................