Method validation of 15 phytochemicals inHypericum lysimachioidesvar.spathulatumby LC-MS/MS, and fatty acid, essential oil, and aroma profiles with biological activities

Abstract

The aim of the present study was to develop and validate a LC-MS/MS method for quantification of 15 phytochemicals inHypericumspecies. The developed method was fully validated in terms of repeatability (inter-day and intra-day precision), limits of detection and quantification, linearity, recovery and relative standard uncertainty. The developed and validated LC-MS/MS method was applied to determine 15 phytochemicals in the ethanol extracts ofH. lysimachioidesvar.spathulatumaerial parts (HLS-A) and roots (HLS-R). Hyperoside was found to be the major compound in HLS-A and HLS-R ethanol extracts (16,560.3 and 3561.6 mu g analyte/g extract, respectively). According to the results of GC-MS analyses,cis-13,16-docosadienoic acid (35.0%), caryophyllene oxide (24.33%) and undecane (28.21%) were determined as the major components in fatty acid, essential oil and aroma compositions ofH. lysimachioidesvar.spathulatum, respectively. HLS-A and HLS-R ethanol extracts showed moderate activity in ABTS cation radical decolorization assay. The major components (hyperoside, astragalin, and quercetin) present in HLS-A and HLS-R ethanol extracts were found to have also the highest antioxidant effect in ABTS cation radical scavenging method. The extracts, main constituents in the extracts and essential oil had no toxic-cytotoxic potential against PDF, MCF-7 and HT-29 cell lines. HLS-A ethanol extract and essential oil exhibited high butyrylcholinesterase inhibitory activity. Quercetin showed the highest inhibitory effect against acetyl- and butyryl-cholinesterase, urease and tyrosinase among the tested samples. According to biological activity studies,H. lysimachioidesvar.spathulatumand their major components might be promoted as promising sources of natural agents and used in the development of nutraceuticals, functional food ingredients and pharmaceutical industry.