Journal Pre-proof Traditional uses, phytochemistry and pharmacology of Chios mastic gum (Pistacia lentiscus var. Chia, Anacardiaceae): A review Vasiliki K. Pachi, Eleni V. Mikropoulou, Petros Gkiouvetidis, Konstantinos Siafakas, Aikaterini Argyropoulou, Apostolis Angelis, Sofia Mitakou, Maria Halabalaki PII: S0378-8741(19)33117-4 DOI: https://doi.org/10.1016/j.jep.2019.112485 Reference: JEP 112485 To appear in: Journal of Ethnopharmacology Received Date: 4 August 2019 Revised Date: 12 November 2019 Accepted Date: 13 December 2019 Please cite this article as: Pachi, V.K., Mikropoulou, E.V., Gkiouvetidis, P., Siafakas, K., Argyropoulou, A., Angelis, A., Mitakou, S., Halabalaki, M., Traditional uses, phytochemistry and pharmacology of Chios mastic gum (Pistacia lentiscus var. Chia, Anacardiaceae): A review, Journal of Ethnopharmacology (2020), doi: https://doi.org/10.1016/j.jep.2019.112485. This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2019 Published by Elsevier B.V. 1 Traditional uses, phytochemistry and pharmacology of 2 Chios mastic gum ( Pistacia lentiscus var. Chia, 3 Anacardiaceae): A review 4 5 Authors: Vasiliki K Pachi, Eleni V. Mikropoulou, Petros Gkiouvetidis, Konstantinos Siafakas, 6 Aikaterini Argyropoulou, Apostolis Angelis, Sofia Mitakou, Maria Halabalaki* 7 8 Department of Pharmacognosy and Natural Products Chemistry, Faculty of Pharmacy, National 9 and Kapodistrian University of Athens, Panepistimiopolis Zografou 15771, Athens, Greece 10 11 Corresponding author: Maria Halabalaki, e-mail: [email protected] 12 1 13 Table of Contents 14 Abbreviations ........................................................................................................................................ 3 15 Abstract ................................................................................................................................................. 4 16 1. Introduction ....................................................................................................................................... 6 17 2. Materials and methods ...................................................................................................................... 9 18 3. Ethnonopharmacological aspects .................................................................................................... 11 19 4. Chemical analysis of Chios Mastic Gum ......................................................................................... 16 20 4.1 Extraction, isolation and identification of CMG constituents ................................................. 16 21 4.1.1 Essential oil and volatile compounds ..................................................................................... 18 22 4.1.2 Extraction and structure elucidation of 1,4-poly-β-myrcene polymer ..................................... 19 23 4.1.3 Isolation and identification of CMG triterpenes ..................................................................... 21 24 4.1.4 Other compounds .................................................................................................................. 30 25 4.2 Analysis of CGM and quality aspects ...................................................................................... 30 26 5. Biological properties ........................................................................................................................ 33 27 5.1 Antioxidant activity .................................................................................................................. 33 28 5.2 Antimicrobial and antifungal properties ................................................................................. 36 29 5.3 Antiinflammatory activity ........................................................................................................ 40 30 5.4 Chemopreventive activity ......................................................................................................... 42 31 5.5 Cardioprotective activity .......................................................................................................... 44 32 5.6 Wound Healing ......................................................................................................................... 46 33 5.7 Other properties ....................................................................................................................... 47 34 6. Pharmacokinetics/ pharmacodynamics .......................................................................................... 49 35 7. Current uses and products .............................................................................................................. 50 36 8. Conclusions and future perspectives ............................................................................................... 56 37 Acknowledgements .............................................................................................................................. 59 38 Conflict of Interest ............................................................................................................................... 59 39 References ............................................................................................................................................ 60 40 2 41 Abbreviations 42 CMG, Chios Mastic Gum; MG, Mastic Gum of unspecified origin; EMA, European Medicines 43 Agency; CMGA: Chios Mastiha Growers Association; PDO, Protected Designation of Origin; 44 TME, Total Mastic Extract; TMEWP, Total Mastic Extract Without Polymer; AMF, Acidic 45 Mastic Fraction; NMF, Neutral Mastic Fraction; TCE, Total Colophony Extract; CMW, Chios 46 Mastic Water; CMO, Chios Mastic Oil; SFE, Supercritical Fluid Extraction; SPE, Solid Phase 47 Extraction; SEC, Size Exclusion Chromatography; DCM, Dichloromethane; MeOH, Methanol; 48 EA, Ethyl Acetate; DE, Diethyl Ether; GC-MS, Gass Chromatography – Mass Spectrometry; 49 GC-FID, Gass Chromatography coupled to a Flame Ionization Detector; RP HPLC, Reverse 50 Phase High Performance Liquid Chromatography; TLC, Thin Layer Chromatography; IMNA, 51 Isomasticadienonic Acid; IMLA, Isomasticadienolic Acid; MNA, Masticadienonic Acid; OA, 52 Oleanolic Acid; MA, Moronic Acid; NMR, Nuclear Magnetic Resonance; MS, Mass 53 Spectrometry; HRMS/MS, High Resolution Tandem Mass Spectrometry; LDL, Low Density 54 Lipoproteins; AAs, Amino Acids; MBC ,Minimum Bactericidal Concentration 55 3 56 Abstract 57 Ethnopharmacological relevance: Chios mastic gum constitutes a unique Greek product, 58 produced exclusively in the southern part of the island of Chios. References about its use from 59 local populations for the treatment of gastrointestinal disorders or as a cosmetic agent can even 60 be encountered in ancient texts of Galen, Theophrastus and Dioscorides. Nowadays, this 61 versatile resin has been rediscovered, not only as a traditional remedy and aromatic agent, but as 62 a potent phytotherapeutic product with various biological properties. 63 Aim of the study: The aim of this study is to quote the summation of the ethnopharmacology, 64 phytochemical profile and pharmacological properties of the resin of Pistacia lentiscus var. Chia 65 and thus provide the scientific community with a summary of the research conducted so far. 66 Furthermore, perspectives and uses are being discussed and studied so as to broaden the field of 67 its applications. 68 Materials and Methods: A comprehensive review of the literature on Pistacia lentiscus var. Chia 69 was performed using as resources scientific databases such as Scopus, Sciencedirect, Pubmed 70 and Web of science, studies and traditional books provided by the Chios Mastiha Growers 71 Association as well as PhD and Master’ s theses. 72 Results: Chios mastic gum has been used as a traditional medicine over the last 2500 years. More 73 than 120 chemical compounds have been identified in the resin and the major components are a 74 natural polymer, acidic and neutral triterpenes and volatile secondary metabolites. Several plant 75 extracts and compounds have been studied for their antibacterial, anti-inflammatory, antioxidant, 76 anti-ulcer, anti-diabetic, cardioprotective and anti-cancer properties in vitro and in vivo . Clinical 77 interventions and trials have also showed the therapeutic potential of Chios mastic gum. In 2015 4 78 Pistacia lentiscus L., resin (mastic) was recognized as a herbal medicinal product with traditional 79 use by the European Medicines Agency (EMA) with two therapeutic indications (mild dyspeptic 80 disorders & skin inflammation/ healing of minor wounds). Over the last years, Chios mastic gum 81 is widely involved in medicinal products, food supplements and cosmetics and has become 82 object of study, also in the field of Pharmacotechnology. 83 Conclusions: Chios mastic’s beneficial properties have been demonstrated in the treatment of 84 gastrointestinal disorders, wound healing, skin inflammations, plasma lipid and blood sugar 85 reduction and oral care. These properties are attributed to triterpenes