UPLC-PDA法测定不同茎色黄芪中花青素的含量及变化

Determination of Anthocyanins Content in Different Colored Stems Astragalus by UPLC-PDA

采用超高效液相色谱法(UPLC-PDA)测定不同茎色黄芪中矢车菊素、飞燕草素的含量,比较不同采收期、放置期黄芪药材中矢车菊素、飞燕草素含量的变化。采用Waters AcQuity UPLC■BEH-C18(100 mm×2.1 mm,1.7μm)色谱柱,流动相为乙腈-0.1%甲酸水溶液,梯度洗脱,流速0.3 mL/min,柱温30℃,检测波长530 nm。矢车菊素、飞燕草素线性范围分别为3.58~28.64μg/mL(r=0.9998)、2.17~17.36μg/mL(r=0.9997),平均回收率(n=6)分别为98.57%(RSD=2.6%)和99.53%(RSD=0.65%)。不同采收期紫茎黄芪中矢车菊素和飞燕草素含量差异较大,但六月下旬后矢车菊素和飞燕草素的总含量基本稳定,药材放置4个月后飞燕草素和矢车菊素含量下降率分别为26.77%和21.36%。绿茎黄芪中未检测到矢车菊素和飞燕草素。方法简便快捷,适用于黄芪中花青素的含量测定研究,为黄芪花青素合成相关基因分析及黄芪药材栽培技术的改进提供了依据。

To establish an UPLC-PDA method for the determination of cyanidin and delphinidin in Astragalus of different colored stems from different gathering and storage periods.Waters AcQuity UPLC■BEH-C18(100 mm×2.1 mm,1.7μm)was adoped with a mobile phase of acetonitrile-0.1%formic acid water.Separation was carried out with a gradient program at a flow rate of 0.3 mL/min,column temperature 30℃and detection wavelength 530 nm.Cyanidin and delphinidin showed good linearity in the ranges of 3.58~28.64μg/mL(r=0.9998)and 2.17~17.36μg/mL(r=0.9997)with average recoveries of 98.57%(RSD=2.6%)and 99.53%(RSD=0.65%),respectively.The contents of cyanidin and delphinidin in Astragalus of purple stems from different gathering periods marked difference,but in late June,the total contents of cyanidin and delphinidin were stable,and four months after harvesting,the decrease rate of the content of delphinidin and cyanidin were 26.77%and 21.36%,respectively.Cyanidin and delphinidin were not detected in Astragalus of green stems.The method is simple,rapid for determination the contents of anthocyanins in Astragalus,and can be used for analyzing the synthetic genes of anthocyanins and improving the cultivation techniques of Astragalus.