University of the Pacific Scholarly Commons University of the Pacific Theses and Dissertations Graduate School 1967 Studies on the essential oil of Anemopsis Californica Ramesh Narmadashanker Acharya University of the Pacific Follow this and additional works at: https://scholarlycommons.pacific.edu/uop_etds Part of the Pharmacy and Pharmaceutical Sciences Commons Recommended Citation Acharya, Ramesh Narmadashanker. (1967). Studies on the essential oil of Anemopsis Californica. University of the Pacific, Thesis. https://scholarlycommons.pacific.edu/uop_etds/400 This Thesis is brought to you for free and open access by the Graduate School at Scholarly Commons. It has been accepted for inclusion in University of the Pacific Theses and Dissertations by an authorized administrator of Scholarly Commons. For more information, please contact [email protected]. STUDIES ON THE ESSENTIAL OIL OF ANEMOPSIS CALIFORNICA A Thesis Presented to the Graduate School University of the Pacific -----------· In Partial Fulfillment of the Require~ents f or the Degree Mast er of Science in Pharmacognosy by Rarnesh Na r~a i as hank e r Acharya May 1967 This thesis, written and submitted by is approved for recommendation to the Graduate Council, University of the Pacific. Department Chairman or Dean: ./ J ~~ (_./' -~ ~ / /}----o~--C'-~-'<- \ Thesis Committee: Dated ACK N O\~LEDGm1ENTS The author wishes to express his sincere gratitude to Dr . ChalJbal for his encouragement a nd guidance throughout this wor k . Without Dr . Cha ubal ' s assistance , thi s work could not have been done . Sincere thanks a re also exnressed to Dr . Roscoe f or his helo in this wo r k . A gift of some authentic sarn ! ~ l es from M/S Fri t zsche Br ot her s Inc ., New Yor k is gr a tefully ac kno t•rl edged . Fi nancial assistance from the Pfeiffer Founda tion and from the University of the Pacific i s 1eepl y ao n rec i a t e~ . TABLE OF CONTENTS Page Acknowledgement ii List of Tables iv List of Figures v I. Introduction 1 II. Extraction and Physical-Chemical Constants of Oil of Anemopsis 4 III. Functional Group Separation of Oil of Anemopsis 8 IV. Fractional Distillation of Oil of Anemopsis 21 V. Chromatographic Methods Introduction 24 Thin-Layer Chromatography 26 Thin-Layer Chromatography of Oil of Anemopsls 29 Gas Chromatography 38 Gas-Liquid Chromatography of Oil of Anemopsis 47 VI. Summary 67 VII. Recommendations for Further Research 68 Bibliography 69 Appendix I 74 LIST OF TABLES TABLE Page 1. Phys ical a nd Ch em ica l Constants of Oil of An emoosis 7 2. Ii.entity of Unknown l? henolic Fr a ction with Thymol 11 3. Fractional Distilla tion of Oi l of An emousis 23 4. Spray Reagents of Thin-La yer Chrornato­ grauhy 32 Gas-Liqui1 Chromatogr aphic ?roce1ure 50 6. Re tention Time of Ma jor ?eaks of Oil of Anemoo s ts l.J·9 Concentra tion of Major Co~~ onents of Oil of Anernops is 66 LIS1' OF F'IGUHES FIGURE Page 1. Functiona l Group Separa tion of Oil of Anemons is 9 2. Infra - re1 Soectra of Thymol and Unknown Phenolic comoounl 1 2 3. Thi n-layer chr0matogram of oil of £1-n emo p~_l.§. 33 4. Thin-la yer chro~ata g ra~ of Phenoli c Fracti on 35 5. Thin-laye r chroma togram of Ca rbonyl Fraction 36 6. Thin-la yer chromatogram of Fraction from Fractional Distill ation of Oil of 37 7. Schematic Diagram of Ga s -Liquid Chro~a t og ranh 39 8 . Gas -Liquid Chr omatogram of oil o f An e~oos is us:i.ng FFAP as a sta tionary ohase ------ - 51 Gas-liquid Chroma togram of oil of Ane~onsis us ing Hy :"'~ erose s?..:: so as a s t a ti o na :r.·y.~--- · ohase 52 10 . Gas-Li qui1 Chro~ato g ram of Fr action # 1 from Fr acti onal Di s til lation of oil of Anemousi s 54 11. Gas-Liq1.tii Chr ~ >mato g ra :n of Fr a c t i on # 2 from Fracti onal Di s tilla tion of oil of Anemousis 55 .12. Gas-Liquid Chroma t ogr a:n of Fr a ction !f 3 fro~ ?r act i ona J Oi stilJation of o il of --Anemon----s i s 13. Ga s-Li qu i d Chr omatog ra~ of Fraction J 4 from ?ractional Di s tj_lla t i on of o i l of -An---emonsi s- 57 14. Infra-red S~ect ra of Methyleugenol and Peak !f 2 59 vi FIGURE Page 15. I nfra-red SDe ctra of ?. i oeritone and Peak !J 3 62 16A. Infra -red S oectra of Peak # 12 6Lt- 16B. Inf r a - red Spectra of Peak ll 12 65 CHAP1'ER I INTHODUCTION The roots and rhizomes of Anemonsis Californica (Nutt. ) Hook and Arn . (family Saurura c eae )have been used by the early Spanish America ns a nd I ndians to treat a variety of ailments . 1 ' 2 ' 3 ' 4' 5 ' 6 The plant is commonly known as Yerba del Mansa, Yerba Man sa , or Manza. The family Saururaceae is renorted t o consist of s-recies r enorted , belong lng to the genus ~l}_~.i~P-~-1..~?.• 1'he plant i s na t ive t o the South Western Un ited States and t o Northern Mexico . I t i s c ommonJ.y f ound in wet alkaline soils at e l evations b elow 6500 f eet . 5 Its range extends from California , Utah , Arizona and Texas to New Mexico . The :.Jlant is vJell d i stri but ed from Lo we r Sacr amento Val ley, San J oaquin Va lle y , South Coast Ranges , Inyo County t o Southern Cal i fornia . 4 An e11 opsi. s Ca l i_f ornica_ a n:l o ther members of the f a mily Saururaceae are consi.der ei t o have little econom ic3.1 i :noort !":'. nce , anart fro~ the "nati ve" meilcina l uses of Ane~ons i s . A~oPs i.~ i s an herbac eous , DPrenni a l d ico t y l e - doneo~s ~ l an t . The nla nt has bgsaJ. l eaves which are 2 large a nd fleshy. The plant blossoms from May t o Augu s t and has an inflorescence c onsisting of ma ny small per fect a~etalou s flowers on a s oike. The bra cts a t the base o f the spike a re white. The pla nt has pr o~ ine n t r oots and rhizome s wh ich exud e a strong a romatic spicy od o r. The r oots a nd r h i zomes a r e c on s i d ered t o b e a very usef u l remedy f or c u t s a nd br u i ses , s or es and spr a i ns . Anemo os i s has a lso been r eported t o b e usef u l as a d iure tic , in rhe uma tism , a ni as a blood ~ urlf ie r in asthma , dys ent e r y a nd gonorr hoea.2 There a r e only a f ew r e uorts pub l 1sh ed on the chemical investigation of t he p l ant Anemoosis . Hort on a nd Paul? i n 1957 first report ed the pr esence of Methy l­ eugenol as a ma in c onstituent of the essential oil 1erived from this p l a nt. Pickering8 in 1958 , c onf i r med the presence of methyl eugen ol (4-allylveratrol). Ch i lds9 in 1 963 , also c on fir ~e1 the or esence o f methyleugenol and re ~o r ted the oresPnce o f a l e uc oant hocyaniii n in t he pe tr o l eQ~ ether ext ract o f the ul a nt. Th e aut hor obser ved t hat the tot a l an ti spa s~oi ic activity of the plant c o uld n o t be due to ~ethyleugeno l a l on e . Childs and Col e l O have a l so r e ~o r ted on t h e phar- . macol ogi c a l a nd a n t i t umo r pr operties o f t he extract o f the roots and r h i zomes of Ane~oos i s . The paucity o f r eoor ts 3 on chem ical investigations and the r eported antitumor activities of the olant drew our attention to the drug . We ~ecided to investigate the essentia l oil of An e~ oosis Ca lifornica in greater detail . The r esults of our investi­ gation to date are included and described in this thesis . CHA PTEH II EX.TriACT ION AND PHYSICAL-CHEfiliCAL CONS'TANTS OF OIL OF ANEMOPSIS Crude Drug Material The crude drug Yerba Mansa i s commercially avail­ able . It i s used at pr ~ s e nt in «native « Mexican medicine . We obtainAd the dried whole roots and rhiz o~es of Anernoo sis Ca lifornica from M/S Ha thaway Al lied Proiucts (2024 West­ gate Av enue , Los Angel es 25 , Ca lifornia ) in t ~ r ee batches . These batches wer e supolied as (i) Lot- RM - 65- 167 , (11) Lot-RM- 65-1029 a nd (iii) L ~ t - RM - 6 7 - 9 1 .