Differentiation of Various Snake Bile Derived from Different Genus by High-Performance Thin-Layer Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry

Background: Snake bile originates from nearly 20 species from three families (Elapidae, Colubridae, and Viperidae). However, the components of various snake bile were not compared with one another. Objecive: The aim of the study was to develop a TLC-MS method for differentiation of various snake bile derived from a different genus. Method: As a type of traditional identification method for Chinese herbs, TLC has considerable advantages as a multicomponent separation system. Results: In this paper, TLC was used to separate the components in snake bile. It was found that the snake bile from Colubridae family and Cobra species is different from the others using TLC. The molecular formulas of the bands were elucidated with TLC coupled with quadrupole–time-of-flight–MS by the TLC-MS interface. Two bands on the plate were identified with the reference substance, and the other three bands were analyzed by the TLC-MS method without the reference substance. The corresponding molecular formulas of these bands were given according to accurate molecular weights. Conclusions: The results from this study indicate that the proposed method is reliable, and it has been successfully applied to the identification of snake bile samples.