Rapid Analysis of Lignans from Leaves of Podophyllum peltatum L. Samples using UPLC-UV-MS

Abstract

The American mayapple (Podophyllum peltatum L.) is a rhizomatous perennial species found throughout the wooded landscape of the eastern half of North America. The American species P. peltatum might be considered as an alternative commercially viable source for podophyllotoxin production. Podophyllotoxin is a natural lignan that is being used as a precursor for the semi-synthetic anti-cancer drugs etoposide, teniposide and etopophos [1]. A new rapid UPLC-UV-MS method has been developed for the analysis of four lignans (4'-O-demethylpodophyllotoxin, podophyllotoxin, α-peltatin and β-peltatin) in P. peltatum L. The chromatographic separation was achieved using a reversed phase C18 column with a mobile phase of water and acetonitrile, both containing 0.05% formic acid. Within 3.0 minutes four main lignans could be separated with detection limits of 0.1, 0.3, 0.3 and 0.2µg/mL, respectively. 4'-O-demethylpodophyllotoxin and α-peltatin appeared most prominently among the lignans obtained. Analyses of mayapple leaves collected from different colonies within a single site indicated a significant variation in 4'-O-demethylpodophyllotoxin, α-peltatin, podophyllotoxin and β-peltatin content. The podophyllotoxin content was found in the range of 0.004–0.77% from the 16 different locations from where the samples were collected. The content of podophyllotoxin is directly proportional to the content of 4'-O-demethylpodophyllotoxin and inversely proportional to α-peltatin and β-peltatin content. LC-mass spectrometry coupled with electrospray ionization (ESI) interface method is described for the identification of four lignans in various populations of plant samples. By applying PCA and HCA, Podophyllum samples collected from various locations were distinguished.