黄烷-3-醇类化合物及其代谢产物的靶向定量方法研究
Development of Target Quantitative Methods of Flavan-3-ols and their Metabolites
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摘要: 黄烷-3-醇类化合物作为重要的膳食多酚，其生物利用度不仅局限于母体化合物，更重要的是其经过体内肠道微生物代谢后的产物。为了明确体内代谢情况及代谢产物对生物利用度的贡献，本研究探究了黄烷-3-醇类化合物及其代谢产物的靶向定量分析方法。根据各化合物子离子响应值的差异，确定其在混合标准溶液中适合的浓度，并建立了超高效液相色谱-三重四极杆-串联质谱（UHPLC-QQQ-MS）定量分析方法，在15 min分析时间内所有化合物的分离度良好，并有适宜的响应值。方法学验证结果表明，定量化合物具有良好的线性关系；并以尿液、粪便和血浆为研究材料考察方法的稳定性，日内和日间精密度RSD值均在15%以下，基质效应介于±20%之间，回收率在70%~120%之间。该方法快速、灵敏、准确、稳定，可用于黄烷-3-醇类化合物在体内的靶向定量研究。Abstract: Flavan-3-ols are important dietary polyphenol, their bioavailability are not only limited to the parent compound, but also the metabolites after the intestinal microbial metabolism in vivo. Aimed to identify the in vivo metabolism and the contribution of metabolites to bioavailability, the quantitative analysis methods of dietary flavan-3-ols and their metabolites were researched. According to the difference response values of the product ions of each compound, the appropriate concentration in the mixed standard was determined, and the quantitative analysis method of ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UHPLC-QQQ-MS) was established. All compounds had good resolution and appropriate response values within analysis time of 15 min. The method validation results showed that most of the quantitative compounds had good linearity. Urine, feces and plasma were used as samples to investigate the intra-day and inter-day precision of the method. The RSD value of the method was below 15%, the matrix effect was within ±20%, and the recovery was mainly between 70% and 120%. The method is fast, sensitive, accurate and stable, which can be used as a method for targeted quantitative research of flavan-3-ol compounds in vivo.
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