This study reports the development and application of a liquid chromatography method coupled to electrospray tandem mass spectrometry (LC-MS/MS) for the identification and quantification of the five most common juvenile hormone (JH) homologs and methyl farnesoate (MF). The protocol allows the simultaneous analysis in a single LC run of JH I, JH II, JH III, JH III bisepoxide (JHB3) and JH III skipped bisepoxide (JHSB3). The identification of JHs is based on multiple reaction monitoring (MRM), using two of the most abundant fragmentation transitions for each hormone. Addition of deuterated JH III as an internal standard permits the absolute quantification of the different JHs. The JH homologs common structural features led to similar chromatographic behavior, as well as related fragmentation patterns, which facilitated the simultaneous detection of all the homologs in a single LC-MS/MS run. The protocol detects JHs in the low femtomole range, allowing often the analysis of JH in individual insects. Fragmentation of each of the JH homologs generates unique diagnostic ions that permitted the identification and quantification of JHs from samples of different species of Diptera, Lepidoptera, Heteroptera and Hymenoptera. Having a simple protocol, which can undisputedly determine the identity of the homologs present in a particular species, provides us with the opportunity to identify and quantify JHs existing in insects that are pests, vector of diseases or important research models.

Department of Biology Florida International University Miami USA

Department of Biology Florida International University Miami USA; Biomolecular Science Institute Florida International University Miami USA

Department of Biology Florida International University Miami USA; Institute of Parasitology Biology Centre CAS Ceske Budejovice Czech Republic

Department of Chemistry and Biochemistry Florida International University Miami USA

Department of Chemistry and Biochemistry Florida International University Miami USA; Biomolecular Science Institute Florida International University Miami USA

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Gut-to-brain regulation of Drosophila aging through neuropeptide F, insulin, and juvenile hormone

Gut-to-brain regulation of Drosophila aging through neuropeptide F, insulin and juvenile hormone

The embryonic role of juvenile hormone in the firebrat, Thermobia domestica, reveals its function before its involvement in metamorphosis

Juvenile hormones direct primordial germ cell migration to the embryonic gonad

The embryonic role of juvenile hormone in the firebrat, Thermobia domestica, reveals its function before its involvement in metamorphosis

Effects of mating on female reproductive physiology in the insect model, Rhodnius prolixus, a vector of the causative parasite of Chagas disease

Female reproductive dormancy in Drosophila is regulated by DH31-producing neurons projecting into the corpus allatum

Crosstalk between Nutrition, Insulin, Juvenile Hormone, and Ecdysteroid Signaling in the Classical Insect Model, Rhodnius prolixus

Genetics tools for corpora allata specific gene expression in Aedes aegypti mosquitoes

Role of Methoprene-tolerant in the regulation of oogenesis in Dipetalogaster maxima

Epoxidation of juvenile hormone was a key innovation improving insect reproductive fitness

Approaches and Tools to Study the Roles of Juvenile Hormones in Controlling Insect Biology

The juvenile hormone described in Rhodnius prolixus by Wigglesworth is juvenile hormone III skipped bisepoxide