PHARMAC E UT I CAL A PPL ICAT IONS OF A PINYON OLEORESIN by VICTOR H. DUKE A thesis submitted to the faculty of the University of Utah in partial fulfillment of the requirements for the degree of Doctor of Philosophy Department of Pharmacognosy College of Pharmacy University of Utah May, 1961 LIBRARY UNIVERSITY elF UTAH I I This Thesis for the Ph. D. degree by Victor H. Duke has been approved by Reader, Supervisory Head, Major Department iii Acknowledgements The author wishes to acknowledge his gratitude to each of the following: To Dr. L. David Hiner, his Dean, counselor, and friend, who suggested the problem and encouraged its completion. To Dr. Ewart A. Swinyard, critical advisor and respected teacher, for inspiring his original interest in pharmacology. To Dr. Irving B. McNulty and Dr. Robert K. Vickery, true gentlemen of the botanical world, for patiently intro­ ducing him to its wonders. To Dr. Robert V. Peterson, an amiable faculty con­ sultant, for his unstinting assistance. To his wife, Shirley and to his children, who have worked with him, worried with him, and who now have succeeded with him. i v TABLE OF CONTENTS Page I. INTRODUCTION 1 II. REPORTED USES OF PINYON OLEORESI N 6 A. Internal Uses 6 B. External Uses 9 III. GENUS PINUS 1 3 A. Introduction 13 B. Pinyon Pines 14 1. Pinus edulis Engelm 18 2. Pinus monophylla Torr. and Frem. 23 3. Anatomy 27 (a) Leaves 27 (b) Bark 27 (c) Wood 30 IV. COLLECTION OF THE OLEORESIN 36 A. Ch ip Method 40 B. Scrape Method 43 C. Bore Method 44 V. REFINEMENT 51 VI. ANALYSIS 57 A. Analysis of the Oil 57 1. Assay for Oil Content 57 2. Purification of Oil 59 3. Yields 60 B. Analysis of the Resin 63 1. Acid Number 63 2. Saponification Number 64 3. Fusion Points 65 (a) Drop Softening Point 65 (b) Melting Point 65 v Page C. Physical Properties of the Purified Oil 67 1. Specific Gravity 67 2. Solubility in Alcohol 68 3. Refractive Index 69 4. Optical Rotation 70 5. Organoleptic Properties 71 6. Non-Volatile Residue 72 VII. PHARMACEUTICAL PREPARATIONS 74 A. Preparations for Local Application 74 1 . W-a t-e r-r--e-m ov eI b I eOleo res i n Ointment 75 2. Oleaginous Oleoresin Ointment 76 3. Cerates 77 4. Dental Cavity Liners 78 B. Preparations for Internal Use 80 1. Emulsions of Oleoresin 80 (a) Liquified Pinyon 82 (b) Primary Emulsion 82 (c) Final Product 83 2. Capsules of Oleoresin 83 3. Water, Aromatic 85 VIII. DISCUSSION 87 A. General 87 B. Collection, Refinement, and Analysis 89 C. Preparations 91 D. Proposed Evaluations 94 IX. SUMMARY AND CONCLUSIONS 95 X. REFERENCES 98 v i LIST OF TABLES Tab I e Page 1 • Bore and Vial, and Cup and Gutter Collection 49 2. Oil Content of Oleoresins 58 3. Acid Number of Resins 64 4. Saponification Number of Resins 64 5. Drop Softening Point of Resins 66 6. Melting Point of Resins 66 7. Specific Gravity of Ojls 68 8. Refractory Index of Oils 70 9. Optical Rotation of Oils 7 1 10. Organoleptic Properties of Oils 73 1 1 . Summary of Volatile Oil Properties 92 1 2. Summary of Resin Properties 92 vii LIST OF PLATES Plate No. Page 1 • Pinus edulis Engelm 2 1 2. Pinus monophylla Torr. and Frem. 26 3. Leaves of Pinyon 29 4. Bark of Pinyon 32 5. Wood of Pinyon 35 6. Chipped bark and limb cross-section of Pinyon 38 7. Cup and Gutter Collection, and Natural Exudate of Oleoresin 42 8. Artificial Scarification of Pinyon 45 9. Artificial Scarification of Pinyon after Twenty Months 46 10. Bore Hole, With and Without Collection Vial 48 1 1 . Cleavenger Oil Trap, Oleoresin Screening Apparatus 56 vii i LIST OF FIGURES Figure No. Pa ge 1. Range of Pinyon 1 7 2. Flow Diagram of Pinyon Oleoresin Extraction and Purification Procedure 6 1 I. INTRODUCTION The pinyon or nut pine comprises a major portion of the Pygmy forest flora of the south-western section of the United States. Tens of thousands of acres of pinyon pines occur in continuous belts. Indeed, in fhe Navajo Indian Reservation alone, there has been estimated to exist over three million acres of pinyon-juniper woodland (Clark and Melis,1953). The Indians and Mexicans have utilized this readily available tree for centuries. The nuts have been used by them as a dietary staple, particularly during the winter months (Peattie, 1953). Early settlers, trappers, and explorers are known to have sought out the pinyon nut as a necessary supple­ ment to their otherwise meager ration (Fremont, 1845). Pinyon nuts have since been shown to be a good source of protein, carbohydrate and fat (Long, 1941). If reft unshelled, these small seeds may be kept for over a year after roasting without becoming rancid (Peattie, 1953). The In d ian s h a vel 0 n gut i I j-·z edt he co u I kin g pro per tie s of pinyon oleoresin. The Apaches made their baskets water­ tight with it. The Navajo still do the same with their water - 2 - bottles, and also use it to make a black dye for their wool. The wood of the pinyon has been used as a source of fuel. The structural timber used in the famed pit houses of the south- western Indians was also pinyon. From a study of the growth rings of this wood, it has been possible to date these pre-historic A mer i can hom e s, and p I ace the i r b u i I· din gas. Ion gag 0 a s 4 0 0 - 900 A. D. (Peattie, 1953). The importance of the pinyon pine to the Indian during the period of intrusion by the first whites was pointed out by John Muir (1912). He states, "This •.. is the Indians' own tree, and many a white man have they killed for cutting it down. II There have been severar studies devoted to the develop- ment of possible commercial uses of western pinyon pines. The latest of these was concerned with determining whether suffi­ cient volatile oil could be obtained from Utah'pinyon pines to be of commercial value as an agent for perfuming pharmaceuti­ cal and cosmetic preparations, and to determine the physical and organoleptic properties of such oils (Cole, 1956). Just previous to this work, and os a part of the t'surveys and resource~ ll studies , The Bureau of Indian Affairs commissioned Deaver and - 3 - Haskell, through the University of Arizona, to conduct a study o nth e pin yon res 0 u r c e son the Na v a i 0 and Hop i I n d ian res e r v a - tions of Arizona and Utah. This work was confined to possible economic exploitation of the oleoresin, resin, and turpentine of Pinus edulis Engelm. as a competitor of the south-eastern pine for the naval stores market (Deaver and Haskell, 1955). The possibility of using western pines as a source of naval stores products was investigated by the United States Depart- men t of A g ric u I t u reF 0 res t S e r vic e ( Bet t s, 1 9 1 2; S c h 0 r g e r 1 9 1 5 ) . Pinus ponderosa, var. scopulorum Engelm., Pinus ponderosa Laws. and Pinus edulis were studied. Using the hack and cup method of collection, Pinus edulis was found to yield an average of 0.147 pounds per cup per week, with the best yield occurring during the months of August and September. The turpentine con­ tent averaging 20.6°k of the tota I oleoresin collected. The tur­ pen tin e was f 0 u n d t 0 con t a i n m 0 s t I y pin e n e, and a Iso. DB a not l­ oil of quite different properties U which were not describedo Schorger (1913) described the oleoresin as having an odor charac teristic of Thuja leaf oil. Cannon (1953) reported on a correlation shown to exist between uranium content in the needles and peeled twigs of piny - 4 - pines and known ore deposits, thus pinyon has been used as a means of locating uranium ore. The oleoresin, commonly known as pinyon gum, has been reported as having been used by both Indians and early white settlers for many and varied medicinal purposes. The medical lore, passed down by word of mouth, indicates that the oleoresin has been successfully used locally as an antiseptic, rubifacient, c 0 u n t e r i r r ita nt, as a d raw i n g sa I v e, and a s a po u I tic e for too t h - aches. The "gumU has been used internally both as a stool- softening laxative and as a treatment for various other gastro­ intestinal disorders. In view of the numerous forms in which pinyon oleoresin has been utilized medicinally, it was thought important to in­ corporate the oleoresin and resin, and the oils of the hybrid described by Cole (1956) into various classes of pharmaceutical and dental products. Pharmaceutically elegant preparations of pinyon products could be prepared for the number of persons currently using the crude materials, and perhaps an even greater market might be developed. If a sufficient demand could be made for these new prepa­ rations as well as demonstrating a superiority of pinyon resin, oil - 5 - and b leo res i n i n est a b lis h e d for m u I a e., the n per hap s the vas t natural pinyon resources of'the West could be developed for some economic advantage .