www.tjprc.org editor@tjprc.org International Journal of Applied, Physical and Bio-Chemistry Research (IJAPBCR) ISSN(P): 2277-4793; ISSN(E): 2319-4448 Vol. 5, Issue 3, Dec 2015, 1-8 © TJPRC Pvt. Ltd. IDENTIFICATION AND QUANTIFICATION OF PODOPHYLLOTOXIN FROM PODOPHYLLUM HEXANDRUM BY ESI-LC/MS/MS SHARMA D.K & KUMAR A Department of Chemistry Himachal Pradesh University, Shimla, Himachal Pradesh, India ABSTRACT An ESI-LC/MS/MS method for the quantification of podophyllotoxin from Podophyllum hexandrum population from Sangla valley of Kinnaur, Himachal Pradesh, India in the Northern Himalayas was developed. Qualification of podophyllotoxin was performed by thin layer chromatography (TLC) with R f values of 0.85 (leaf) and 0.94 (root) when compared with the standard podophyllotoxin. UV-VIS spectrophotometeric studies showed electronic absorption at λ max value 284 nm. Gradient chromatographic separation of Podophyllotoxin was performed on a YMC-Pak Pro C-18 : 4.6mm × 250 mm, HPLC column, packed with 1.5 μm particles equipped with a 0.7m pre-filter (Supleco), using mobile phase methanol/water (1/1, v/v) and 100% methanol, both containing 0.1% ammonium hydroxide (25%) and 10 m mol ammonium acetate (pH 9). Structural information from the tandem mass spectrometric data compared with those of lignin markers already reported for the herb established the quantitative high yield of pophyllotoxin. The method could be employed in the absence of reference standards for the marker and was particularly useful in view of the scarcity of the chemical standard of podophyllotoxin. KEYWORDS: Podophyllotoxin; Podophyllum Hexandrun; ESI-LC/MS/MS; Tandem Mass Spectrometry Received: Oct 05, 2015; Accepted: Oct 12, 2015; Published: Oct 18, 2015; Paper Id.: IJAPBCRDEC20151 INTRODUCTION Podophyllum hexandrum also known as Indian Mayapple, which is an endangered medicinal perennial herb is belonging to the family Berberidaceae. Podophyllum hexandrum, a moisture and shade loving erect, glabrous, succulent herb thriving from Himalayan zone at an altitude between 1300 to 4300 m above from sea level. Extensive chemical investigation of Podophyllum species revealed presence of a number of compounds like podophyllin, podophyllotoxin, querectin, 4-dimethypodophyllotoxin, kaempherol, picropodophyllotoxin, α-peltatin and β-peltatin [1]. The podophyllotoxin content in Indian Podophyllum is more (7-15%) in comparison to other species notably Podophyllum peltatum (4-8%), the most common species in American sub-continent [2]. Recently podophyllotoxin has acquired great importance and high medicinal status due to its effectiveness as antimitotic, anticancer and immunostimulatory activity [3,4,5] especially for curing uterine tumors [6]. The rhizome powder of the plant is used as laxative or to get rid of intestinal worms and also used as poultice to treat warts and tumorous growth on the skin. Semi synthetic derivatives of podophyllotoxin e.g. etoposide (VP-16) [7], etopophos [8] and tenoposide (VM-26) are effective against the treatment of lung cancer, a variety of leukaemia, and other solid tumours [9]. Recently, Podophyllum hexandrum extracts have been reported to offer radioprotection by modulating free radical flux involving the role of lignans presents [10-16]. Due to its anticancerous property podophyllotoxin is in Original Article