背景:塞洛通(SLT)是由人参提取物(人参C.A.Meyer的干燥根和根茎)制成的标准化草药配方,银杏(银杏叶),和藏红花(番红花的污名)。遵循补气活血的中医原则,可兼容治疗血管性痴呆(VaD)。人参在中国被广泛用作恢复力量的补品。银杏和藏红花传统上长期被用作药物,其主要作用是活血化瘀。
目的:现有的药理和临床证据证明SLT是治疗VaD的一种有前途的药物。了解配方草药及其有效成分如何配合产生综合效果,本研究旨在建立一种高灵敏度和准确的定量方法来揭示人类SLT的血浆暴露情况。
方法:通过使用在线SPE进行样品提取,随后进行LC-MS/MS测定,实现了人血浆中总共17种SLT衍生组分的多重定量。其中,8人参皂苷(Rg1,Re,F1,Rf,Rb1,Rb2,Rc和Rd)在ESI模式下测定,和银杏黄酮槲皮素,山奈酚,异鼠李素处于ESI模式。通过优化样品提取和LC分离的条件,提高了灵敏度,以及质量参数。4种银杏内酯,包括银杏内酯A,B,C和白果内酯,和2种藏红花素1及其代谢产物藏红花素,在负离子模式下同时分析,一系列保护解决方案确保了它们的稳定性。
结果:所有人参皂苷的定量下限在0.078ng/mL时非常敏感,银杏黄酮0.033-0.2ng/mL,对于银杏内酯为0.75或1.5ng/mL,对于Crocins为3ng/mL。方法的准确性和精密度得到充分验证,2名健康志愿者的临床样本分析证明了其适用性。
结论:所开发的方法将有助于进一步详细的临床药代动力学研究,以阐明SLT的作用机制并制定其合理的治疗方案。
BACKGROUND: 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.
OBJECTIVE: 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.
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.
CONCLUSIONS: 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.