14-羟基芸苔素甾醇高效液相色谱分析方法

Quantitative Determination of 14-Hydroxylated Brassinosteroid by HPLC

[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。

[Aims ] A rapid and efficient high performance liquid chromatography method for determination of14-hydroxylated brassinosteroid was developed. [Methods ] The samples were treated by derivatization reaction with1-Naphthaleneboronic acid as derivatization reagent and with methanol as solvent. The determination was carried out on Eclipse XDB C_（18） column with the mixture of acetonitrile and water（60 ∶40, by vol） as mobile phase and flow rate of 1.0 m L/min at 30 ℃. [Results ] The complete separation of the main composition and impurities was achieved in10 min under optimized conditions. When the mass concentration of 14-hydroxylated brassinosteroid ranged from 25.0 to400.8 mg/L, the linear relationship between peak area and concentration was well and the linear coefficient correlation was 0.9997. The relative standard deviation（RSD） for seven replicate determination was 0.55% and the mean recovery was 100.80%. [Conclusions ] The method is efficient, accurate and easy, which provides reliable method for determination and quality control of 14-hydroxylated brassinosteroid and other brassinosteroids.