保幼激素及其代谢产物的HPLC分离方法的改进和应用

Modification and application of a high performance liquid chromatography method to separate juvenile hormones and their metabolites

传统的正相高效液相色谱 (normalphasehighperformanceliquidchromatography,NP HPLC)可以较好地分离保幼激素 (ju venilehormone ,JH)Ⅰ、Ⅱ和Ⅲ ,但不能分离保幼激素的代谢产物及其类似物。经过改进的反相高效液相色谱 (reversephasehighperformanceliquidchromatography,RP HPLC)不仅可以很好地分离保幼激素 ,还能定性定量分析保幼激素的代谢产物及其类似物。离体培养昆虫咽侧体 (corporaallata ,CA)所合成的、被同位素标记的痕量保幼激素可以用以上两种色谱方法进行分离和鉴定。此外 ,RP HPLC还可以用来分离体内或体外同位素标记的保幼激素代谢产物 ,以及测量血淋巴中的保幼激素滴度。

Juvenile hormones (JH) Ⅰ,Ⅱ and Ⅲ can be effectively separated by traditional normal phase high performance liquid chromatography (NP-HPLC), but their metabolites and analogues can not be clearly separated by this method. We have recently modified a method of reverse phase high performance liquid chromatography (RP-HPLC). This modified method can be used to separate not only different juvenile hormones, but also a variety of juvenile hormone metabolites and analogues, qualitatively and quantitatively. Trace amounts of radiolabeled juvenile hormone, which was synthesized and secreted by the in vitro incubated corpora allata (CA), could be separated by both NP-HPLC and RP-HPLC methods. Moreover, the RP-HPLC can be used to separate radiolabeled juvenile hormone metabolites and analogues, in both in vivo and in vitro conditions, as well as to measure juvenile hormone levels in the hemolymph.