气相色谱-质谱法分析测定甲壳类动物中的大甲基法尼酯

大甲基法尼酯是甲壳类动物体内的一种激素,在早期发育阶段具有改变形态的生物活性,并对繁殖起调控作用.本论文发展了一种基于正相高效液相色谱法纯化、气相色谱-质谱联用仪分析的综合分析方法.以非生物活性异构体反式-顺式-大甲基法尼酯(C-T-MF)作为内标物并用无分流注射法进样,采用电子撞击电离源电离后检测.利用无分流注射模式检测时,气相色谱-质谱联用仪的T-T-MF检出下限为100 pg/(次.柱).若取血淋巴样品4 mL,从最终25μL提取物中注入2μL,此方法的检出下限为0.3 ng/mL.在样品制备第一步(包括溶剂萃取、高效液相色谱纯化、气相色谱-质谱联用仪分析)就加入25 ng T-T-MF的条件下,总回收率为79%(RSD=5%,n=5).这种综合分析方法已被成功地应用于测定甲壳类动物血淋巴中T-T-MF的含量,如龙虾(Homarus americanus)、墨吉尔虾(Fennero peneaus merg-uiensis)、小龙虾(Procambarus clarkii).

Quantification of Methyl Farnesoate Levels in Crustacea by Gas Chromatography-Mass Spectrometry Analysis

Methyl farnesoate (MF) is a hormone known to have important morphogenetic bioactivity in early developmental stages as well as having regulatory roles in crustacean reproduction. A comprehensive analytical method based on normal -phase, high performance liquid chromatography purification and gas chromatography- mass spectrometry (GC- MS) analysis has been developed for the quantitative determination of trans -trans -MF (T -T -MF) in hemolymph of crustaceans. Detection was done by electron ionization with splitless injection using the non- biological isomer, cis- trans- MF (C -T- MF), as internal standard. GC -MS instrument detection limit for the T- T- MF was 100 pg on -column using the splitless injection mode. The detection limit of the method was 0.3 ng/mL using 4 mL hemolymph samples and injections of 2 μL of the final 25 μL extract. The total recovery at 25 ng T - T - MF when added at the first step (solvent extraction, HPLC purification and GC/MS analysis) was 79% (RSD =5%, n =5). The validated method was successfully applied to the determination of T - T - MF levels in crustaceans, such as lobsters Homarus americanus, banana prawns Fennero peneaus merguiensis and crayfish Procambarus clarkii.