title: Quantitative determination of juvenile hormone III and 20-hydroxyecdysone in queen larvae and drone pupae of Apis mellifera by ultrasonic-assisted extraction and liquid chromatography with electrospray ionization tandem mass spectrometry.
abstract: In this paper, a method for the rapid and sensitive analysis of juvenile hormone III (JH III) and 20-hydroxyecdysone (20E) in queen larvae and drone pupae samples was presented. Ultrasound-assisted extraction provided a significant shortening of the leaching time for the extraction of JH III and 20E and satisfactory sensitivity as compared to the conventional shake extraction procedure. After extraction, determination was carried out by liquid chromatography-tandem mass spectrometry (LC-MS/MS) operating in electrospray ionization positive ion mode via multiple reaction monitoring (MRM) without any clean-up step prior to analysis. A linear gradient consisting of (A) water containing 0.1% formic acid and (B) acetonitrile containing 0.1% formic acid, and a ZORBAX SB-Aq column (100 mm × 2.1 mm, 3.5 μm) were employed to obtain the best resolution of the target analytes. The method was validated for linearity, limit of quantification, recovery, matrix effects, precision and stability. Drone pupae samples were found to contain 20E at concentrations of 18.0 ± 0.1 ng/g (mean ± SD) and JH III was detected at concentrations of 0.20 ± 0.06 ng/g (mean ± SD) in queen larvae samples. This validated method provided some practical information for the actual content of JH III and 20E in queen larvae and drone pupae samples.
authors.affiliation: Bee Research Institute, Chinese Academy of Agricultural Sciences, PR China.
authors.forename: Jinhui
authors.initials: J
authors.lastname: Zhou
authors.forename: Yitao
authors.initials: Y
authors.lastname: Qi
authors.forename: Yali
authors.initials: Y
authors.lastname: Hou
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Xue
authors.forename: Liming
authors.initials: L
authors.lastname: Wu
authors.forename: Jinzhen
authors.initials: J
authors.lastname: Zhang
authors.forename: Fang
authors.initials: F
authors.lastname: Chen
mesh_terms: D000818:Animals; D001516:Bees / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004441:Ecdysterone / Q000032:analysis* / Q000302:isolation & purification; D007814:Larva / Q000737:chemistry*; D016018:Least-Squares Analysis; D016014:Linear Models; D011679:Pupa / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012717:Sesquiterpenes / Q000032:analysis* / Q000302:isolation & purification; D013010:Sonication; D021241:Spectrometry, Mass, Electrospray Ionization; D013696:Temperature
chemical_list: D012717:Sesquiterpenes; D004441:Ecdysterone; C036585:juvenile hormone III