A partition assay for the simultaneous determination of insect juvenile hormone esterase and epoxide hydrolase activity.
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.
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Department of Entomology, North Carolina State University, Raleigh 27695-7613.
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D000096:Acetone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000032:analysis* / Q000037:antagonists & inhibitors; D002855:Chromatography, Thin Layer; D004851:Epoxide Hydrolases / Q000032:analysis*; D007313:Insecta / Q000201:enzymology*; D000432:Methanol; D009795:Octanes; D012995:Solubility
D009795:Octanes; D000096:Acetone; C055697:3-octylthio-1,1,1-trifluoro-2-propanone; D002265:Carboxylic Ester Hydrolases; C038003:juvenile hormone esterase; D004851:Epoxide Hydrolases; C055698:juvenile hormone epoxide hydrolase; C045798:2,2,4-trimethylpentane; D000432:Methanol