摘要
目的建立大鼠尿液中蜜炙黄芪三种活性成分毛蕊异黄酮、芒柄花素和黄芪甲苷的含量测定方法。方法采用Phenomenex kinetex柱为色谱柱,0.2%乙酸水-乙腈为流动相梯度洗脱,柱温2 5℃,流速0.3 ml/min,在负离子模式下采用UPLC/Q-TOF-MS技术对大鼠尿液中毛蕊异黄酮、芒柄花素和黄芪甲苷的含量进行测定。结果毛蕊异黄酮、芒柄花素和黄芪甲苷分别在6.42 min,10.48 min和11.14 min左右出峰,表明三个物质在色谱柱上分离度良好;其分子离子峰分别为m/z 283.06[M-H]^-,m/z 267.06[M-H]^-和m/z 843.47[M+CH_3COO]^-,选择性强。毛蕊异黄酮、芒柄花素和黄芪甲苷分别在1.55~250μg/ml,0.234~50.0μg/ml和0.100~12.8μg/ml范围内线性良好;在不同浓度内,此法的日内、日间精密度和准确度均小于±15%,相对回收率均大于80%,基质效应的RSD值均小于15%。结论 UPLC/QTOF-MS技术可用于蜜炙黄芪的体内含量测定,且符合生物样品分析要求,灵敏度高,重复性好,是一种快速可靠的新方法。
Objective To establish a method for the determination of calycosin, formononetin and astragaloside IV in rat urine af- ter oral administration of honey - processed Astragalus. Methods The residues were separated on a Phenomenex kinetex column with the mobile phase consisting of acetonitrile - water with 0.2% acetic acid at the flow rate of 0.3 mL/min. The column tem- perature was set at 25 ℃ and the contents of calycosin, formononetin and astragalus 1V in rat urine were determined by UPLC/Q - TOF - MS in the negative ion mode. Results The results showed that the separation of calycosin, formononetin and astragaloside IV were good at 6.42 min, 10.48 min and 11.14 rain, respectively. The selective molecular ion peaks were m/z 283.06[ M - HI -, m/z 267.06[ M - HI - and m/z 843.47[ M ＋ CHACO0] -, respectively. The linear ranges of calyeosin, formononetin and astragaloside IV were 1.55 -250 μg/mL, 0. 234 -50.0 μg/mL and 0. 100 - 12.8 μg/mL , respectively. Within the different concentration levels, the intra - day and inter - day precision and accuracy were less than ± 15% , the relative recovery was more than 80% , and the matrix effect of RSD was less than 15%. Conclusion UPLC/Q - TOF - MS technology might be used for the determination of the compositions of honey - processed Astragalus in vivo. It is a quick and reliable method for the analysis of bio- logical samples with high sensitivity and good repeatability.
出处
《时珍国医国药》
CAS
CSCD
北大核心
2017年第11期2564-2567,共4页
Lishizhen Medicine and Materia Medica Research
基金
国家自然科学基金(81202917
81573607)
