Simultaneous determination of naphtodianthrones, emodin, skyrin and new bisanthrones in Hypericum perforatwn L. in vitro shoot cultures

摘要

Hypericum perforatum L. (St John's wort) extracts are top-selling herbal medicines for the treatment of various ailments including mild to moderate depression across the world. One of the major bioactive compounds of these extracts is hypericin that is also considered as a lead molecule for drug development. Here, we report on the development of a robust method for the simultaneous quantification of hypericins (hypericin, pseudohypericin, protohypericin and protopseudohypericin), their presumed precursors (emodin and skyrin) and new skyrin derivatives (oxyskyrin, iridoskyrin, rubroskyrin and luteoskyrin). Compounds extracted from the in vitro shoot cultures of H. perforatum L. in ethanol, acetone, 80 % methanol, ethylacetate and dichloromethane were separated by ultra-performance liquid chromatography (UPLC), quantified by photo diode array/fluorescence (PDA-FLR) and annotated by Q- Exactive Orbitrap mass spectrometer (UPLC-QE-MS). Analyses of the UPLC data revealed that ethanol is the best extractant for the efficient quantification of all target compounds, although all of them could also be extracted in acetone and 80 % methanol. The extractants as well as the method of detection proved to be important for the efficient quantification of naphtodianthrones. Our results are expected to significantly contribute to the dissection of the hypericin biosynthesis pathway and the quality control of Hypericum extracts in the pharmaceutical industry.