Modification and application of a high performance liquid chromatography method to separate juvenile hormones and their metabolites
OUYANG Ying-Chun1, LI Sheng 2*(1. Institute of Zoology, Chinese Academy of Sciences, Beijing 100080, China; 2. Department of Biological Sciences, Illinois State University, Normal, IL 61790-4120, USA)
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.