超声辅助-分散液液微萃取结合超高效液相色谱——串联质谱分析马尿中痕量氨鲁米特

Ultrasonic Assisted Dispersive Liquid-Liquid Microextraction Combined with Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry for the Determination of Trace Aminolutamide in Equine Urine

建立了一种简单、快速、高灵敏检测马尿中氨鲁米特的超声辅助分散液液微萃取/超高效液相色谱-串联质谱方法。将3mL马尿用1mol/L氨水调节至pH 10.5,以邻二氯苯为萃取剂,四氢呋喃为分散剂,配比为4∶5,组成微萃取体系共0.9mL进行提取,经过1min频率为80Hz的超声处理,4℃下以4000r/min高速离心10min后获得提取液,在40℃下氮气吹干后,经1mL 0.1%甲酸-乙腈(95:5)溶解后使用超高效液相色谱-三重四极杆串联质谱检测,以甲喹酮为内标,正离子电喷雾下多反应监测模式进行分析。结果显示,样品溶液在0.1~50ng/mL线性关系良好(r=0.9994),方法的检出限为0.05ng/mL,定量限为0.1ng/mL。尿样在三种加标浓度下的回收率为90.2%~109.7%,RSD不高于5.6%且基质效应低。经方法学验证,该方法可用于马术运动兴奋剂检测中痕量氨鲁米特的分析。

A simple,rapid and highly sensitive method for the determination of aminolumide in horse urine by ultrasonic assisted dispersion liquid-liquid microextraction/ultra high performance liquid chromatography tandem mass spectrometry was established.3ml of horse urine was adjusted to pH 10.5 with 1 mol/L ammonia,o-dichlorobenzene as extractant and tetrahydrofuran as dispersant,with a ratio of 4:5.A total of 0.9ml of microextraction system was formed for extraction.After ultrasonic treatment with a frequency of 80Hz for 1 min and high-speed centrifugation at 4000R/min at 4℃for 10min,the extract was obtained.After being dried with nitrogen at 40°C,it was dried with 1ml of 0.5ml 1%the dissolution of formic acid acetonitrile(95:5)was detected by ultra performance liquid chromatography with three quadrupole tandem mass spectrometry.The results showed that the sample solution had good linearity in the range of 0.1~50ng/mL by using the method of multiple reaction monitoring under positive electrospray ionization.(r=0.9994),the detection limit of the method is 0.05ng/ml and the quantitative limit is 0.1ng/ml.the recovery of urine samples under three spiked concentrations is 90.2%-109.7%,RSD is not higher than 5.6%and matrix effect is low.The method can be used for the analysis of trace aminolumide in the detection of equestrian sports stimulants.