Multiplex quantitation of 17 drug-derived components in human plasma after administration of a fixed herbal preparation of Sailuotong using combined online SPE-LC-MS/MS methods

Abstract

Ethnopharmacological relevance: Sailuotong (SLT) is a standardized herbal medicine formula made from extracts of ginseng (the dried root and rhizome of Panax ginseng C. A. Meyer), ginkgo (the leaves of Ginkgo biloba L.), and saffron (the stigma of Crocus sativus L.). It is prescribed compatibly for the treatment of vascular dementia (VaD) following the TCM principle of Qi-invigorating and Blood-activating. Ginseng is widely used as a tonic for the restoration of strength in China. Ginkgo and saffron have been traditionally used for a long time as medicines with the main effect of promoting blood circulation and removing blood stasis.

Aim of the study: SLT has been proven to be a promising medicine for VaD by existing pharmacological and clinical evidence. To understand how the formula herbs and their active ingredients cooperate to produce comprehensive effects, the present study aimed to establish a highly sensitive and accurate quantitative method to reveal the plasma exposure profile of SLT in humans.

Material and methods: Multiplex quantitation of a total of 17 SLT-derived components in human plasma was fulfilled by using online SPE for sample extractions followed by LC-MS/MS determinations. Among them, 8 ginsenoside (Rg1, Re, F1, Rf, Rb1, Rb2, Rc and Rd) were determined in ESI+ mode, and ginkgo flavonoids of quercetin, kaempferol, isorhamnetin were in ESI- mode. Improved sensitivity was achieved through optimizing the condition of sample extraction and LC separation, as well as mass parameters. 4 ginkgolides, including ginkgolide A, B, C and bilobalide, and 2 crocins of crocin-1 and its metabolite crocetin, were analyzed concurrently in negative ion mode, and their stability was ensured by a series of protective solutions.

Results: The lower limit of quantitation was achieved to be extremely sensitive at 0.078 ng/mL for all ginsenosides, 0.033 ‒ 0.2 ng/mL for ginkgo flavonoids, 0.75 or 1.5 ng/mL for ginkgolides and 3 ng/mL for crocins. The methods were fully validated to be accurate and precise, and applicability was demonstrated by the analysis of clinical samples from 2 healthy volunteers.

Conclusion: The developed methods should be useful in further detailed clinical pharmacokinetic research for clarifying the effect mechanism of SLT and formulating its rational therapeutic regimens.

Keywords: Crocus sativus; Ginkgo biloba; Human plasma; Online-SPE-LC-MS/MS; Panax ginseng; Sailuotong.