Abstract—The propose of this study was to investigate in vitro thrombolytic activity of Zingiber cassumunar Roxb. and Prasaplai, a Thai herbal formulation of Z. cassumunar Roxb. Herbs were extracted with boiling water and concentrated by lyophilization. To observe their thrombolytic potential, an in vitro clot lysis method was applied where streptokinase and sterile distilled water were used as positive and negative controls, respectively. Crude aqueous extracts from Z. cassumunar Roxb. and Prasaplai formula showed significant thrombolytic activity by clot lysis of 17.90% and 25.21%, respectively, compared to the negative control water (5.16%) while the standard streptokinase revealed 64.78% clot lysis. These findings suggest that Z. cassumunar Roxb. exhibits moderate thrombolytic activity and cloud play an important role in the thrombolytic properties of Prasaplai formula. However, further study should be done to observe in vivo clot dissolving potential and to isolate active component(s) of these extracts. Keywords—Aqueous extract, prasaplai formula, thrombolytic activity, Zingiber cassumunar Roxb. I. INTRODUCTION LOOD clot formation, thrombosis, has been a severe problem of the blood circulation system. Thrombus or embolus obstructs the blood flow by blocking the blood vessel therefore depriving blood and oxygen supply to tissue, leading to tissue necrosis. Atherothrombotic diseases such as acute myocardial or cerebral infarction and stroke are serious consequences of the thrombus formed in blood vessels. Usually thrombolytic agents such as streptokinase (SK), urokinase (UK), or tissue-plasminogen activator (t-PA) [1]- [3], are used to dissolve the formed clots in the vessels, however, these drugs have certain limitations which cause severe and sometime fatal disorders including systemic fibrinolysis, anaphylactic reaction, and bleeding tendency [4]- [6]. Traditional herbs have been used since ancient times to treat many diseases. Herbs are often known as safe because they are natural products. Previous studies have shown that many herbs possessed antithrombotic activity [7]-[11]. For instance, K. W., J. K., E. P., and T. S. are with the Department of Thai Traditional Medicine, Thai Traditional Medicine College, Rajamangala University of Technology Thanyaburi, Pathum Thani, 12130, Thailand (phone: +66 25921999; fax: +66 25921900; e-mail: warachate_k@rmutt.ac.th, khemjira_j@rmutt.ac.th, pattaranut_e@rmutt.ac.th, surachai_te@rmutt.ac.th). S. S. is with the Department of Medical Technology, School of Allied Health Sciences and Public Health, Walailak University, Nakhon Si Thammarat, 80160, Thailand (phone: +66 75672104-5; fax: +66 75672106; e- mail: suriyan.su@wu.ac.th). methanolic extract of Umbilicaria esculenta exhibited both antithrombotic activity in vivo and in vitro [7]. However, herbs that could be used for thrombolysis has been few reported. Ayurvedic Prasaplai is a Thai traditional herbal drug for pain treatment. Prasaplai is derived from “Prasa” and “Plai” in which Prasa means 50% in amounts and Plai is a Thai name of Zingiber cassumunar Roxb. Prasaplai formula contains 50% Z. cassumunar Roxb., while the remaining ingredients consist of equal amounts of each plant including the root of Calamus (Acorus calamus L.), the peel of Kaffir lime (Citrus hystrix DC.), the bulb of Waanhomdaeng (Eleutherine americana (Abul.) Merr.), the bulb of Garlic (Allium sativum L.), the fruit of Long pepper (Piper retrofractum Vahl.), the fruit of Black pepper (Piper nigrum L.), the rhizome of Zedoary (Curcuma zedoaria Roscoe.), the rhizome of Ginger (Zingiber officinale Roxb.), the seed of Black cumin (Nigella sativa L.), and two chemical compounds, sodium chloride and camphor [12]. Usually, it was used for pain relief during menstruation dysmenorrheal [13]. This traditional Thai drug has been described by alternative medicine for treatment of primary dysmenorrheal [14]. Z. cassumunar Roxb, a species of plant in the Zingiberaceae family, is a major ingredient in Prasaplai formula. It is cultivated throughout Southeast Asia [15] and widely used as a Thai traditional herb for treatment of inflammation [16], muscular and joint pain [17], rheumatoid arthritis [18], skin diseases [19], asthmatic symptoms [20], [21], abscesses, wound healing, and menstrual disorders, as well as in food in Thailand [22]. The chemical compositions of the rhizome of Z. cassumunar Roxb. have been studied and revealed that there are phenylbutenoids [23], [24], cassumunaquinones [25], β- sesquiphellandrene [26], cassumunarins [27], [28], β- sitosterol, terpinen-4-ol [29], [30], triquinacene 1,4-bis (methoxy), (E)-1-(3,4-dimethoxyphenyl)buta-l,3-diene, (E)-1- (3,4-dimethoxyphenyl) but-1-ene, and (Z)-ocimene [19]. To date, no studies have investigated the thrombolytic properties of Z. cassumunar Roxb. and its formula, Prasaplai. With the aim to evaluate thrombolytic activity of the aqueous extracts from Z. cassumunar Roxb. and Prasaplai formula, we investigated the thrombolytic activity by using in vitro model. II. MATERIALS AND METHODS A. Preparation and Extraction of Plant Material The rhizome of Z. cassumunar Roxb. and the 9 raw herbs composing the Prasaplai formula were purchased from a Evaluation of Thrombolytic Activity of Zingiber cassumunar Roxb. and Thai Herbal Prasaplai Formula Warachate Khobjai, Suriyan Sukati, Khemjira Jarmkom, Pattaranut Eakwaropas, Surachai Techaoei B World Academy of Science, Engineering and Technology International Journal of Pharmacological and Pharmaceutical Sciences Vol:11, No:5, 2017 286 International Scholarly and Scientific Research & Innovation 11(5) 2017 ISNI:0000000091950263 Open Science Index, Pharmacological and Pharmaceutical Sciences Vol:11, No:5, 2017 publications.waset.org/10007201/pdf