THE PENNSYLVANIA STATE UNIVERSITY SCHREYER HONORS COLLEGE DEPARTMENT OF TOXICOLOGY NATIVE AMERICAN ETHNOBOTANY AND TOXICOLOGICAL ANALYSIS OF THE ZANTHOXYLUM GENUS KAYLEE SIMON SPRING 2021 A thesis submitted in partial fulfillment of the requirements for a baccalaureate degree in Pharmacology & Toxicology with honors in Pharmacology & Toxicology Reviewed and approved* by the following: Joshua Kellogg Assistant Professor in the Agricultural Sciences Thesis Supervisor Gary H. Perdew H. Thomas and Dorothy Willits Hallowell Chair in Agricultural Sciences Honors Advisor * Electronic approvals are on file. i ABSTRACT Zanthoxylum americanum, otherwise known as northern prickly ash, is an aromatic shrub or small tree native to North America. It is used in several traditional medicines to cure various diseases. This shrub has been known to act as a dermatological aid, antifungal, toothache remedy, and a cold and cough remedy1. A wide range of chemical compounds found within the plant have shown promising pharmacological characteristics. These include coumarins, alkaloids, and furanocoumarins which come from the stem, root, bark, berries, and many other parts of the plant. Some of the most important furanocoumarins found in Z. americanum extracts are psoralen, 8-MOP, and imperatorin, and it is these compounds that give the fruit, husk, and leaf extracts their broad spectrum of antifungal activity2. Furanocoumarins are phototoxic by covalently binding to DNA forming adducts which allow Z. americanum to be used in phototherapeutic applications against fungal infections2,3. Furanocoumarins also correlate to the antimicrobial activity of Z. americanum2. The chemical compounds within Zanthoxylum americanum make it a common therapeutic for many infections and other skin conditions. Documenting and analyzing how other species within the Zanthoxylum genus can be used in traditional medicine is important to further understand the properties that cause toxicity or have therapeutic effects. This review documents the ethnobotany, chemistry, and toxicity/bioactivity of other species within the Zanthoxylum genus including Z. armatum DC. and Z. acanthopodium DC. ii TABLE OF CONTENTS LIST OF FIGURES ....................................................................................................................... iii LIST OF TABLES ......................................................................................................................... iv ACKNOWLEDGEMENTS ............................................................................................................ v INTRODUCTION .......................................................................................................................... 1 ZANTHOXYLUM ARMATUM DC. ............................................................................................ 2 ETHNOBOTANY ...................................................................................................................... 2 CHEMISTRY ............................................................................................................................. 4 TOXICITY/BIOLOGICAL ACTIVITY .................................................................................... 5 ZANTHOXYLUM ACANTHOPODIUM DC. .............................................................................. 7 ETHNOBOTANY ...................................................................................................................... 7 CHEMISTRY ............................................................................................................................. 8 TOXICITY/BIOLOGICAL ACTIVITY .................................................................................. 10 CONCLUSION ............................................................................................................................. 13 BIBLIOGRAPHY ......................................................................................................................... 14 iii LIST OF FIGURES Figure 1: Zanthoxylum armatum. (a) A mature flowering plant (b) Young fruits (c) Ripe fruits. Reconfigured from Phuyal et al. (2019)11. ...................................................................................... 2 Figure 2: Terpene Structures in Z. armatum DC14 .......................................................................... 4 Figure 3: Flavonoid Structure in Z. armatum DC 11 ....................................................................... 4 Figure 4: Zanthoxylum acanthopodium DC. fruits. Reconfigured from (“Zanthoxylum acanthopodium DC.”, 2020)25. ........................................................................................................ 7 iv LIST OF TABLES Table 1: Ethnomedicinal uses of Zanthoxylum armatum. Reconfigured from Phuyal et al. (2019)11 ........................................................................................................................................... 3 Table 2: Bioactivity of Zanthoxylum armatum and extracts. Reconfigured from Brijwal et al. (2013)16. .......................................................................................................................................... 6 Table 3: Identified chemical content of Zanthoxylum acanthopodium fruit. Reconfigured from Sibero et al. (2020)31. ...................................................................................................................... 9 v ACKNOWLEDGEMENTS I would like to thank Dr. Joshua Kellogg for believing in me and allowing me to work in his laboratory. It has been a great journey seeing the development of the lab over these past few semesters. Giving me the opportunity to conduct as much research as possible given the COVID-19 restrictions has been a great experience. Having him help me every step of the way in the thesis writing process has been greatly appreciated. I would also like to thank Dr. Vanden Heuvel for always answering my questions and guiding me through the undergraduate process. I also want to express my gratitude for all of the members of the Kellogg laboratory for being a source of positivity during our lab meetings and in the lab. Last but not least, I would like to thank my parents and friends for watching me grow over these past four years and pushing me to follow my dreams. Without their love and support, I would not be the person I am today. 1 INTRODUCTION The Zanthoxylum genus belongs to the Rutaceae family which consists of over 200 species including trees and shrubs in warm subtropic areas around the world4. Some are more commonly used in Northeastern United States, while others are found in places like India, Nepal, and Indonesia. Zanthoxylum comes from the word Xanthoxylum which in Greek derives to “zanthon xylon” or yellow wood5. Many articles and authors use Zanthoxylum and Xanthoxylum interchangeably5. Species in the Zanthoxylum genus have distinct features. They have the presence of recurved spines on the trunk and branches5. The leaf shapes can be varied having up to 15 pairs of leaflets5. The seeds of the fruits are either red or black with a slight shine5,6. The fruits can contain one to five aromatic carpels5,7. Species in the Zanthoxylum genus have been used in traditional medicine to treat fungal infections, stomach ailments, inflammation, and rheumatic diseases4. They are importance sources of edible fruits, oils, and medicinal plants5. The major components of Zanthoxylum species are furanocoumarins, monoterpenes, and alkaloids. They have shown potential biological activities such as antioxidant, antibacterial, antimicrobial, and antifungal. 2 ZANTHOXYLUM ARMATUM DC. ETHNOBOTANY Zanthoxylum armatum DC. is an erect shrub or small tree found in the hot valleys of the Himalayas and India8. The seeds, bark, and fruit are used to treat various diseases. The fruits, branches, and thorns are used to help elevate toothaches whereas the seeds and bark are used in fever, indigestion, and cholera treatment9,10. There are also some studies showing that Z. armatum DC has the potential to help manage diabetes. Figure 1: Zanthoxylum armatum. (a) A mature flowering plant (b) Young fruits (c) Ripe fruits. Reconfigured from Phuyal et al. (2019)11. 3 Table 1: Ethnomedicinal uses of Zanthoxylum armatum. Reconfigured from Phuyal et al. (2019)11 4 CHEMISTRY Essential oil extractions from Zanthoxylum armatum DC branches and leaves contain �- Terpinene (45.56%), piperitone (33.47%), and 3-carene (8.88%)12, all of which are monoterpenes. However, different places of origin and harvest time seem to cause varying chemical composition for the same plant essential oil12. This is seen with Z. armatum DC as varied harvest times resulted in a change of main components to 2-undecanone (19.75%), followed by 2-tridecanone (11.76%) and �-caryophyllene (9.88%)12,13. Essential oils extracted from Kashmir and Jummu contained linalool, linalyl acetate and limonene14,15.The main constituents of the essential oils of Zanthoxylum armatum DC are consistently monoterpenes which have demonstrated insecticidal as well as antibacterial activities16. It has even been used to treat some cancers 16. Crude methanol extracts of Zanthoxylum armatum DC are seen to also contain phenolic and flavonoid compounds like tambulin that show antimycotic potential11. Figure 2: Terpene Structures in Z. armatum DC14 Figure 3: Flavonoid Structure in Z. armatum DC 11 5 TOXICITY/BIOLOGICAL ACTIVITY Zanthoxylum armatum DC methanolic and ethanolic fruit extracts evidenced antioxidant properties through free radical activity