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.
Conclusions: 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.
[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。.
Keywords: Biomphalaria glabrata; Dopamine; Nervous system; Serotonin; UPLC MS/MS.