Liquid chromatography coupled to ion trap-tandem mass spectrometry to evaluate juvenile hormone III levels in bee hemolymph from Nosema spp. infected colonies

详细信息	查看全文 | 推荐本文 |

• 作者：A.M. Ares^a ; M.J. Nozal^a ; J.L. Bernal^a ; R. Martí ; n-Herná ; ndez^b ; ^c ; M.Higes^b ; J. Bernal^a ; ^{jose.bernal@qa.uva.es}
• 关键词：Juvenile hormone III ; Honey bee hemolymph ; LC&ndash ; MS/MS ; Nosema ceranae ; Nosema apis
• 刊名：Journal of Chromatography B ; Analytical Technologies in the Biomedical and Life Sciences
• 出版年：2012
• 期刊代码：124_15700232
• 类别：ch
• 出版时间：15 June, 2012
• 卷：899
• 期：Complete
• 页码：146-153
• 文件大小：436 K

摘要

It has been described a fast, simple and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to measure juvenile hormone III (JH III), which was used to study of the effects of Nosema spp. infection on JH III levels in bee hemolymph. Honey bee hemolymph was extracted by centrifugation and mixed with a solution of phenylthiourea in methanol. This mixture was then centrifuged and the supernatant removed and evaporated to dryness. The residue was reconstituted in methanol containing the internal standard (methoprene) and injected onto an LC-MS/MS (ion-trap) system coupled to electrospray ionization (ESI) in positive mode. Chromatography was performed on a Synergi Hydro-RP column (4 渭m, 30 mm 脳 4.60 mm i.d.) using a mobile phase of 20 mM ammonium formate and methanol in binary gradient elution mode. The method was fully validated and it was found to be selective, linear from 15 to 14,562 pg/渭L, precise and accurate, with%RSD values below 5%. The limits of detection and quantification were: LOD, 6 pg/渭L; LOQ, 15 pg/渭L. Finally, the proposed LC-MS/MS method was used to analyze JH III levels in the hemolymph of worker honey bees (Apis mellifera iberiensis) experimentally infected with different Nosema spp. (Nosema apis, Spanish and Dutch Nosema ceranae strains). The highest concentrations of JH III were detected in hemolymph from bees infected with Spanish N. ceranae.