title: Fast, ultra-trace detection of juvenile hormone III from mosquitoes using mass spectrometry.
abstract: In the present work, a new protocol for fast separation and quantification of JH III from biological samples using liquid chromatography coupled to electrospray tandem mass spectrometry is described. In particular, the proposed protocol improves existing methodologies by combining a limited number of sample preparation steps with fast LC-MS/MS detection, providing lower limits of detection and demonstrated matrix effect control, together with high inter and intraday reproducibility. A limit of detection of 8pg/mL (0.32pg on column) was achieved, representing a 15-fold gain in sensitivity with respect to previous LC-MS based protocols. The performance of the LC-MS/MS protocol is comparable to previously described JH III quantitation protocol based on fluorescence detection, with the added advantage that quantification is independent of the availability of fluorescent tags that are often unavailable or show quite diverse responses on a batch-to-batch basis. Additionally, a detailed description of the JH III fragmentation pathway is provided for the first time, based on isolation of the molecular ion and their intermediate fragments using in-source MS/MS, MS/MS(n) and FT-ICR MS/MS measurements. The JH III workflow was evaluated as a function of developmental changes, sugar feeding and farnesoic acid stimulation in mosquitoes and can be applied to the detection of other juvenile hormones.
authors.affiliation: Department of Chemistry and Biochemistry, Florida International University, Miami, USA.
authors.forename: Cesar E
authors.initials: CE
authors.lastname: Ramirez
authors.affiliation: Department of Biology, Florida International University, Miami, USA; Biomolecular Science Institute, Florida International University, Miami, USA.
authors.forename: Marcela
authors.initials: M
authors.lastname: Nouzova
authors.affiliation: Department of Chemistry and Biochemistry, Florida International University, Miami, USA.
authors.forename: Paolo
authors.initials: P
authors.lastname: Benigni
authors.affiliation: Department of Chemistry and Biochemistry, Florida International University, Miami, USA.
authors.forename: J Martin E
authors.initials: JME
authors.lastname: Quirke
authors.affiliation: Department of Biology, Florida International University, Miami, USA; Biomolecular Science Institute, Florida International University, Miami, USA.
authors.forename: Fernando G
authors.initials: FG
authors.lastname: Noriega
authors.affiliation: Department of Chemistry and Biochemistry, Florida International University, Miami, USA; Biomolecular Science Institute, Florida International University, Miami, USA. Electronic address: fernandf@fiu.edu.
authors.forename: Francisco
authors.initials: F
authors.lastname: Fernandez-Lima
mesh_terms: D000818:Animals; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002853:Chromatography, Liquid; D009033:Culicidae / Q000737:chemistry*; D057230:Limit of Detection; D015203:Reproducibility of Results; D012717:Sesquiterpenes / Q000032:analysis*; D053719:Tandem Mass Spectrometry
chemical_list: D012717:Sesquiterpenes; C036585:juvenile hormone III
keywords: Juvenile hormone III; Liquid chromatography; MRM; extraction; quantification