UHPLC-DAD-ESI-MS/MS and HPTLC profiling of ash leaf samples from different commercial and natural sources and their in vitro effects on mediators of inflammation.

Introduction: In European traditional medicine, common ash leaf infusion is recommended by European Medicines Agency to treat minor articular pain and to increase the amount of urine for flushing minor urinary complaints. However, a comprehensive ultra-high-performance liquid chromatography diode array detector electrospray ionisation tandem mass spectrometry (UHPLC-DAD-ESI-MS/MS) analysis of this pharmacopeial plant material has never been performed. Moreover, the number of biological and pharmacological investigations proving the usefulness of this plant material in recommended traditional uses is surprisingly small.

Objective: Phytochemical profiling of ash leaf samples from different commercial and natural sources and the determination of the in vitro effects on inflammatory mediators in a model of human neutrophils.

Methods: Ash leaf samples were characterised by total hydroxycinnamic acid content and by high-performance thin layer chromatography (HPTLC), UHPLC-DAD-ESI-MS/MS methods. The effects of leaf infusions on reactive oxygen species (ROS), tumor necrosis factor (TNF-α), interleukin 8 (IL-8), interleukin 1β (IL-1β), and monocyte chemoattractant protein 1 (MCP-1) production by neutrophils were measured using luminol-dependent chemiluminescence and enzyme-linked immunosorbent assay (ELISA).

Results: In ash leaf samples 64 compounds were identified or partly identified together with four unknown compounds. The major compounds detected belong to different structural groups, including phenolic acid derivatives, phenylethanoids, flavonoids, iridoids, secoiridoids and lignans. The major compounds detected in ash samples were chlorogenic acid, quercetin-3-O-rutinoside, verbascoside, oleuropein and ligstroside. However, one sample contained coumarin derivatives. This finding suggested adulteration with other Fraxinus species and/or plant parts. All infusions were able to inhibit ROS, cytokine and chemokine production.

Conclusions: The performed phytochemical and biological analyses contribute to the knowledge about this pharmacopeial plant material and supports its traditional use to treat minor inflammatory complaints.