A Thesis on fhe Seasonal Variation In Santonin Content of the Indigenous Halophytlc Artemislas submitted in partial fulfilment of the requirements for the degree of Doctorate of Philosophy of the University of 01asgovf hy James Goutts* March 1933. ProQuest Number: 13905419 All rights reserved INFORMATION TO ALL USERS The quality of this reproduction is dependent upon the quality of the copy submitted. In the unlikely event that the author did not send a com plete manuscript and there are missing pages, these will be noted. Also, if material had to be removed, a note will indicate the deletion. uest ProQuest 13905419 Published by ProQuest LLC(2019). Copyright of the Dissertation is held by the Author. All rights reserved. This work is protected against unauthorized copying under Title 17, United States C ode Microform Edition © ProQuest LLC. ProQuest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, Ml 48106- 1346 PREFACE* In submitting this thesis my thanks are due to Prof*D*Ellis who kindly acted as supervisor* To Dr* J.W.Gregor of the Scottish Society for Research in Plant Breeding, who supervised the cultivation of plants on the Society’s ground at Corstorphin© and who gave much appreciated advice, I am very grateful* To the late E*M*Holmes, Esq*, Dr*J.P*Todd and Lloyd, I would acknowledge my indebtedness for their ever-ready advice and helpful criticism. Information regarding the localities in which the required plants are to be found, was supplied by Prof*A.W. Bartlett of Durham University; Prof*v«.Grant Craib of Aberdeen University; Prof*R.Lloyd Praeger of Dublin University; Prof.R.A. Robertson of St* Andrews University; ProfiE*J.Salisbury of London University; Prof*J*Small of Belfast University; A.J. Wllmott, Esq* of the British Museum; Miss E.Vachell of Cardiff and others* It is a pleasure to acknowledge my thanks to them* CONTENTS Page Preface • . ♦ • • ill Introduction • • . * X P4RT I. Historical Chapter I. Early History . * * * 5 II. Present Day Sources of Santonin 16 III. Chemistry ♦ • • • • 19 The essential oil; santonin; artemisin. PART IX« Experimental* X?. The Ecology . ♦ * . * 27 V* The Morphology • •*••• 29 Systematic; additional morphological characters • VI* The Soil * * . • • * 40 VII* Methods of Evaluating the Drug • • * 43 Sublimation; colorimetric* VIII. The Quantitative Assay of Santonin * * 61 Observations on the existing polarimetric, volumetric and gravimetric methods; new technique; comparative results with different methods. IX* Some Physical Properties of Santonin * 74 Solubility in benzene; density of the solution; solubility In sodium carbonate solution* X* Determination of the Seasonal Variation in Santonin Content. • .... 83 -V- Chapter Page XX, The Santonin Content of British end Belgian Material • • • • • • , • . 100 Occurrence and santonin content of English and Welsh; Occurrence in Scotland; santonin content of Belgian. XII. A. gallioa under Cultural Conditions . * • 129 XXXI. Physiological Significance of the Occurrence of Santonin , . , , . 152 XIV* Conclusions ♦ • • • * * • . 157 Appendix* Methods for the Assay of Santonin ♦ 142 Keferenees * 150 Figures V - XXVII . * * * * * 154 Map • * « » ~ * • « * . 172 1 ■-.a % v. t . '-vr , ■- -i- V 'V=. ''1 I ‘.i .. ( v;: ' 5-:^'^"' ■>. a.l"' ' ;■ 1 : • ' v ' ■ v ' *.'■ i ' ') ' h , A h tr'i'-: INTRODUCTION Santonin is at present extracted commercially from the unexpanded flowerheada of Artemisia clna. (Berg), Willkomm, and certain other species of Artemisia. This part of the herb is considered to contain the highest proportion of this principle and it is believed to be present in maximum amount Just before the flowerheads expand. It is known, however,that the leaves of certain other species contain santonin and may be used for extraction of it. There is no evidence regarding its occurrence in the leaves of A. olna, or in the flowerheads of the other species. It was thought that the behaviour of santonin might be analogous to that of certain alkaloids, etcetera, which have been shown to be formed in the leaves and to be translocated to the younger parts of the plant, such as the growing points. It was decided therefore to carry out an investigation of the different parts of successive batches of Artemisia maritime,Linn., and A. galllca.Willd., which contain santonin and which are to be found growing wild on the east coast of Scotland. The seasonal variation In the santonin content would thus be ascertained. This was carried on over a period of three seasons and a regular periodic variation was found to exist. A probable explanation of the variation was formulated, correlating it with respiratory processes In the plant. Batches of the two species were also collected from a large number of localities all round the coasts of England and Wales. On examination these were also found to contain santonin, although the proportion was not so high as in the Scottish plant. Wherever possible a few successive batches of the species were obtained from these other localities and a similar seasonal variation In the santonin content was found. Cultivated plants also showed a similar variation. A new assay process was formulated, and was found to be reliable in yielding consistent results and can be applied to drugs of low santonin content. Most of the existing methods were tried out and observations on their use are made. At the beginning of the research it was not known whether the two speoies were sufficiently delimited and Dr. J.W* Gregor of the Scottish society for Research in Plant-Breedingj was asked by the writer for his opinion on the advisability, and desirability for the present research, of carrying out a study of the range of variation within the species of/piants. He replied as follows :~ ttTho experimental study of speoies, and of the ecological and geographical races which make up species, promises ultimately to bring the results of taxonomic studies into line with the requirements of research workers In other fields of science* *Untll such time, however, as the results of the combined efforts of geneticists, ecologists, cytologlsts and morphologists regarding the status of the specific and sub­ specific units of our flora become available the research worker -3- who is desirous of having M s experimental material identified, be it plant or animal, has no alternative but to accept the opinions, based on morphology, of competent systematists. While ^identification by observation” must be relied upon at the present time, it should be remembered that experimentation has shown that races morphologically indistinguishable may be physiologically dissimilar.* "To attempt an analysis of the sub-specific units contained in A. maritime of Linne is obviously outside the scope of your investigation and in my opinion the only course open to you is to accept the diagnosis of the Kew botanists and mention the habitat from which each collection was taken#* The two species are sufficiently well distinguished without referring to the sub-specific characters. This work Is more concerned with the seasonal variation in santonin content, which is probably similar in all similarly situated variants of the species, than with the purely botanical characters of the plant. in accordance with Dr. Gregor*s advice and in view of the futility of an examination of the sub-specific characters of plants from only one or a limited number of localities, a comprehensive study of these characters was not made. Certain characters which have been shown to be of value in identifying certain other species of Artemisia, when the flowerheads alone are available for this purpose, were, however, thoroughly examined. The author*s identification of the two species was confirmed by authorities on systematy, to whom sample batches were sent. CHAPTER I. EARLY HISTORY The botanical name, Artemisia, is a very ancient one. Its origin Is attributed by some authors to Artemisia, wife of Mausolus, and later Queen of Carla, who died in 348 B*C#, and who gave it her own name in honour of its virtues* Others consider it to be derived from Artemis, the Creek name of Diana, Goddess of the Moon, who was regarded as presiding over the diseases of women. The use of the plant as a drug can be definitely traced, under the name of wsantonicaw as far back as the time of Dloscorides (1) in 77 a ,D, He mentions tvfo varieties, one of which grew in Gaul in the country of the Santones, the modern aaintonge, and it seems probable that the name, ^santonica11, was derived from this, Fliny (2) also speaks of the plant as the current remedy for intestinal worms. The Romans during their conquest of Gaul in the first century A,D. found the plant growing there, «nd used it as an effective vermifuge* The species found growing in Saintonge was probably A, galllca, Willd, The second variety mentioned by Dloscorides came from Cappadocia In N.B* Asia Minor and has been regarded through the ages as a Valuable drug* She French plant soon passed into disuse and it is not mentioned as a source of the drug in lat#P literature; nevertheless Its local use has apparently continued* Later references always Indicate a drug coming from further east* Saladinus (3) in 1450 refers to a remarkable vermifuge for ehildren 11 semen eawyg-C t ^ vel Alexandra£feal?t 'il U' M which came from the east, although the geographical source is not definitely stated* The drug has been known by several names. The "semen sanctum” of Saladlnus, is stated by some authors to have been applied to the herb because the priests were wont to carry twigs of thi$ plant in preference to branches of the Olive, in t^ieir ancient religious ceremonies. Others ascribe this name to the fact that it was believed to originate in the Holy Land or Palestine* An­ other name which was much used, "semen contra", is a contraction of the descriptive Latin "semen contra vermes”.