藏羚粪便中类固醇激素的高效液相色谱分析

Analysis of Steroids from Chiru Feces by High Performance Liquid Chromatography

[目的]通过建立粪便中生殖激素含量的测定方法,为研究藏羚羊的繁殖特征奠定基础。[方法]以2-(11H-苯﹝α﹞咔唑)-乙基肼基甲酸酯(BCECH)为柱前衍生化试剂,探讨皮质醇、皮质酮、睾酮和孕酮4种类固醇激素衍生物的最佳色谱分离条件。通过比较4种类固醇激素标准品出峰时间,确定粪样中目标物质峰。[结果]最优衍生条件为:以乙腈为衍生化溶剂,以三氯乙酸为催化剂,衍生化温度65℃,衍生试剂用量为总摩尔类固醇的10倍以上。在建立的色谱条件下,藏羚粪样中4种类固醇激素含量色谱图有较多的杂质峰,表明粪样中的成分很复杂。[结论]该研究建立了藏羚羊粪便中类固醇激素高效液相色谱的分析方法,实现了藏羚粪便中4种类固醇化合物的定量测定。

[Objective]The research aimed to lay a foundation for studying the reproductive characters of chiru through establishing the measurement method of reproductive hormone content in the feces.[Method]Using 2-（11H-benzo ﹝ a ﹞ carbazol-11-yl） ethylcarbonylhydrazine（BCECH） as pre-column derivatization reagent,the optimum chromatographic separation conditions of 4 kinds of steroid hormone ramification（including cortisol,corticosterone,testosterone and progesterone） were discussed.Through comparing the peak time of 4 kinds of standard steroid hormones,the peak of the target substance was confirmed.[Result]The optimum derivatization conditions were as follows：with acetonitrile as derivatization solvent,with trichloroacetic acid as activator,derivatization temperature of 65℃,derivatization solvent dosage was ten times than total moore of steroid.Under the established chromatography conditions,the chromatogram of the content of 4 kinds of steroid hormones had more impurity peaks,showing that the components in the feces samples.[Conclusion]The analysis method of steroid hormone in feces by high performance liquid chromatography was established and the quantitative measurement of 4 kinds of steroid compounds in the feces of chiru was realized.