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[目的]建立高效液相色谱测定14-羟基芸苔素甾醇含量的方法。[方法]以萘硼酸作为衍生化试剂,甲醇作为溶剂进行衍生化反应。采用Eclipse XDB C_(18)色谱柱,以乙腈-水(体积比60∶40)等度洗脱,流速为1.0 mL/min,柱温为30℃进行测定。[结果]14-羟基芸苔素甾醇和杂质在10 min内能达到基线分离。14-羟基芸苔素甾醇在25.0~400.8 mg/L质量浓度范围内,峰面积与质量浓度线性关系良好,回归方程的相关系数为0.9997,7次平行测定的相对标准偏(RSD)为0.55%,平均回收率为100.80%。[结论]该方法简单实用,可为14-羟基芸苔素甾醇及其他芸苔素甾醇的质量控制提供一种准确可行的方法。
[Objective] To establish a method for the determination of 14-hydroxy-brassinosterol by high performance liquid chromatography. [Method] With naphthaleneboronic acid as derivatization reagent and methanol as solvent, the derivatization reaction was carried out. The elution was performed on an Eclipse XDB C_ (18) column with isocratic elution of acetonitrile-water (60:40 by volume) at a flow rate of 1.0 mL / min and a column temperature of 30 ° C. [Result] The 14-hydroxy-brassinosterol and impurities could reach baseline separation within 10 min. The linear relationship between peak area and mass concentration of 14-hydroxy brassinosterol was good in the concentration range of 25.0-400.8 mg / L, the correlation coefficient of regression equation was 0.9997, and the relative standard deviation (RSD) of seven parallel determinations was 0.55% , The average recovery was 100.80%. [Conclusion] The method was simple and practical, which could provide an accurate and feasible method for quality control of 14-hydroxy-brassinosterol and other brassinosteroids.