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采用健康人新鲜粪便提取液与九节龙皂苷I(ADS I)在厌氧条件、37℃下培养72 h,通过液液萃取、旋转蒸发、复溶等方法对样品进行预处理,应用高效液相色谱-蒸发光散射检测法(HPLC-ELSD)和超高效液相色谱-电喷雾-串联质谱检测法(UHPLC-ESI-MS/MS)推断ADS I的生物转化产物结构。结果发现ADS I在人肠道菌作用下主要发生脱糖基反应,初步确定其结构分别为3个次生苷(M1、M2、M3)和1个苷元(M4)。本研究首次利用离体培养人肠道菌群法对ADS I进行生物转化研究,为ADS I在人肠道内转化吸收提供了科学依据。
The fresh manure extract of healthy people and ADS I were incubated under anaerobic conditions at 37°C for 72 h. Samples were pretreated by liquid-liquid extraction, rotary evaporation, reconstitution and other methods. HPLC-ELSD and ultra performance liquid chromatography-electrospray-tandem mass spectrometry (UHPLC-ESI-MS/MS) were used to infer the structure of ADS I biotransformation products. As a result, it was found that ADS I mainly undergoes a deglycosylation reaction under the action of human intestinal bacteria, and its structure is initially determined as three secondary glycosides (Ml, M2, M3) and one aglycon (M4). For the first time, this study used the in vitro culture of human intestinal flora to study the biotransformation of ADS I, which provided a scientific basis for the conversion and absorption of ADS I in the human intestine.