光滑双脐螺神经系统生物胺UPLC MS/MS定量检测方法的建立  

Quantitative determination of biogenic amine from Biomphalaria glabrata nervous system by UPLC MS/MS

作　　者：黄涛 郭云海 刘和香[1] 张仪[1] HUANG Tao;GUO Yun-hai;LIU He-xiang;ZHANG Yi(National Institute of Parasitic Diseases, Chinese Center for Disease Control and Preventio;WHO Collaborating Centre for Tropical Disease;National Center for International Research on Tropical Diseases, Ministry of Science and Technolog;Key Laboratory of Parasite and Vector Biology, National Health and Family Planning Commission, Shanghai 200025, Chin)

机构地区：[1]中国疾病预防控制中心寄生虫病预防控制所世界卫生组织热带病合作中心科技部国家级热带病国际联合研究中心国家卫生和计划生育委员会寄生虫病原与媒介生物学重点实验室,上海200025

出　　处：《中国血吸虫病防治杂志》2018年第2期145-148,178,共5页Chinese Journal of Schistosomiasis Control

基　　金：国家重点研发计划(2016YFC1200501)

摘　　要：目的建立光滑双脐螺神经系统五羟色胺和多巴胺的超高液相色谱-三重四级杆串联质谱(UPLC MS/MS)定量检测方法。方法在纯甲醇溶液中破碎光滑双脐螺神经系统,经过2次高速离心获得含有目标检测物的上清液,通过ACQUITY UPLC BEH Amide色谱柱、Waters TQ-XS系列质谱检测器,采用ESI源、正离子模式上机检测。结果光滑双脐螺神经系统五羟色胺UPLC MS/MS检测限为0.03 ng/ml,定量限为0.1 ng/ml;多巴胺UPLC MS/MS检测限为0.05 ng/ml,定量限为0.15 ng/ml。在1~40 ng/ml添加范围内,五羟色胺的回收率为90.68%~94.72%;多巴胺的回收率为91.68%~96.12%。结论建立的UPLC MS/MS方法操作简单、检测结果稳定、重复性良好,可用于光滑双脐螺神经系统五羟色胺和多巴胺的定量检测分析和研究。Objective To establish a method for the quantitative determination of serotonin and dopamine in the nervous system of Biomphalaria glabrata by using ultra high performance liquid chromatography-tandem quadrupole mass spectrometry（UPLC MS/MS）.Methods The B. glabrata nervous system was broken in the pure methanol solution after obtaining it by dissecting with microscope. Then,the supernatant containing the target substance after twice high speed centrifugation was got. The extraction was separated on an ACQUITY UPLC BEH Amide column with Waters TQ-XS series mass spectrometry detector,with ESI source and positive electrospray ionization mode when the machine testing.Results The detection limit of serotonin was 0.03 ng/ml and the limit of quantification was 0.1 ng/ml. The detection limit of dopamine was 0.05 ng/ml and the limit of quantification was 0.15 ng/ml. The recoveries of serotonin ranged from 90.68% to 94.72% over the range of 1 to 40 ng/ml. The recoveries of dopamine ranged from 91.68% to 96.12% over the range of 1.0 ng/ml to 40 ng/ml.Conclusion The established UPLC MS/MS method is simple,stable and reproducible. It can be used for the quantitative analysis of serotonin and dopamine in the nervous system of B. glabrata snails.

关 键 词：光滑双脐螺 神经系统 五羟色胺 多巴胺 UPLC MS/MS 

分 类 号：R383.24[医药卫生—医学寄生虫学]