A Multiplex GC-MS/MS Analysis for the Quantitative Moni-toring of Bilobalide, Ginkgolides and Ginkgotoxin in Ginkgo biloba-Derived Products and Biomaterials

Abstract: The bioactivity Ginkgo biloba-derived extracts and other preparations is attributed to the presence of secondary metabolites, especially terpene trilactones (ginkgolides and bilobalide), flavonoids but also toxic constituents, like ginkgotoxin. In this study, we set up a multiplex method using a gas chromatography coupled with tandem mass spec-trometry (GC-MS/MS) for the simultaneous quantitative analysis of six of these char-acteristic metabolites, namely ginkgolide A, B, C, J, bilobalide and ginkgotoxin. Param-eters were set up and optimized for multiple reaction monitoring (MRM) to allow the sensitive and selective monitoring of specific collision induced dissociation transitions for each analyte. The method was applied to quantify and compare the above ingredi-ents after derivatization in the methanol extracts of a commercial supplement, ginkgo nuts, ginkgo cells from suspension and callus cultures, and microvesicles (MVs) and nanovesicles (NVs) isolated from ginkgo seeds. As a result of the MRM analysis, the commercial supplement contained 7,77% terpene lactones per tablet. The seed samples contained 50 parts per million (ppm) ginkgolide A and 14 ppm ginkgolide B, but gink-golide C and J were below the limit of quantitation. Ginkgo biloba cells from in vitro cultures, MVs and NVs contained considerably less bilobalide and ginkgolides than the seed and the commercial supplement. Keywords: Ginkgo biloba; extracellular vesicles; GC-MS/MS; derivatization; quantita-tive analysis, multiple reaction monitoring, ginkgolides, bilobalides, ginkgotoxin