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目的:建立高效液相色谱法(HPLC)测定新疆紫草毛状根中乙酰紫草素和β,β′-二甲基丙烯酰紫草色素含量的方法。方法:采用YMCODS-AC18色谱柱(4.6mm×150mm,5μm),流动相为乙腈-水-甲酸(700:300:0.5);检测波长为275nm;流速为1.0mL.min-1;柱温为35℃。结果:β,β′-二甲基丙烯酰紫草素对照品浓度在10.32～206.4μg.mL-1范围内与峰面积线性关系良好,r=0.9999;平均回收率为99.6%,RSD为1.7%(n=9)。乙酰紫草素对照品浓度在51.55～1031.0μg.mL-1范围内,与峰面积线性关系良好,r=0.9999;平均回收率为99.8%,RSD为2.0%(n=9)。优化培养的毛状根中不含从新疆紫草根中分离得到的紫草素;乙酰紫草素的含量明显高于β,β′-二甲基丙烯酰紫草素的含量。结论:高效液相色谱法操作简单快速,重复性好,可用于新疆紫草毛状根中乙酰紫草素和β,β′-二甲基丙烯酰紫草色素含量的测定。
Objective: To establish a high performance liquid chromatography (HPLC) method for the determination of acetylhosulfan and β, β’-dimethacrylocyanine in Xinjiang violet grass hairy roots. Methods: The mobile phase consisted of acetonitrile-water-formic acid (700: 300: 0.5) with a wavelength of 275 nm and a flow rate of 1.0 mL · min-1 on a YMCODS-AC18 column (4.6 mm × 150 mm, 35 ° C. Results: The calibration curve of β, β’-dimethylacrylshikonin had a good linear relationship with the peak area in the range of 10.32 ~ 206.4μg.mL-1, r = 0.9999; the average recovery was 99.6% and the RSD was 1.7 % (n = 9). The calibration curve of acetylshikonin in the range of 51.55 ~ 1031.0μg.mL-1 showed a good linear relationship with the peak area (r = 0.9999). The average recovery was 99.8% and the RSD was 2.0% (n = 9). The cultured hairy roots did not contain shikonin isolated from Xinjiang shikonin; the content of acetylshikonin was significantly higher than that of β, β’-dimethylacrylshikonin. Conclusion: HPLC is simple, rapid and reproducible. It can be used for the determination of acetylschin and β, β’-dimethacryl in pigment of hairy roots of Lithospermum.