Share M R, Roe R M
Department of Entomology, North Carolina State University, Raleigh 27695-7613.
Anal Biochem. 1988 Feb 15;169(1):81-8. doi: 10.1016/0003-2697(88)90257-6.
A partition assay was developed to measure insect juvenile hormone (JH) I and III metabolism in biological samples containing both JH esterase and JH epoxide hydrolase activity. The assay utilizes commercially available radiochain 3H-labeled JH as substrate and the selective JH esterase inhibitor 3-octylthio-1,1,1-trifluoro-2-propanone. JH partitions into an isooctane phase and the metabolites JH acid, JH diol, and JH diol-acid into aqueous methanol after incubation of JH substrate with inhibited and uninhibited sample. The assay provides a time- and cost-efficient alternative to the currently available thin-layer chromatography method for the measurement of JH esterase and epoxide hydrolase activity.
开发了一种分配测定法，用于测量同时具有保幼激素酯酶和保幼激素环氧水解酶活性的生物样品中的昆虫保幼激素（JH）I和III的代谢情况。该测定法使用市售的放射性链3H标记的JH作为底物以及选择性保幼激素酯酶抑制剂3-辛硫基-1,1,1-三氟-2-丙酮。在将JH底物与受抑制和未受抑制的样品孵育后，JH分配到异辛烷相中，而代谢产物JH酸、JH二醇和JH二醇酸则分配到甲醇水溶液中。该测定法为目前用于测量保幼激素酯酶和环氧水解酶活性的薄层色谱法提供了一种省时且经济高效的替代方法。
