超高效液相色谱-质谱法分析人全血、肝脏中维库溴铵

Determination of vecuronium bromide in whole blood and blank liver with UPLC-MS/MS

目的:建立超高效液相色谱-质谱法(UPLC-MS/MS法)测定人血液、肝脏中维库溴铵。方法:取空白血、空白肝(经匀浆)加入一定量维库溴铵标准溶液,加入pH 8氨水稀释后涡旋离心,取上清液过混合型弱阳离子交换柱(WCX)进行固相萃取提取纯化。提取液采用UPLC-MS/MS检测。色谱分离采用UPLC~?BEH HILIC色谱柱。流动相体系:A相为乙腈,B相为5 mmol·L^(-1)甲酸铵水溶液(甲酸调pH 3.5左右),梯度淋洗;质谱检测采用正离子扫描,多反应离子监测模式(MRM)。检测结果以维库溴铵母离子m/z 557.526和子离子m/z 100.103、356.346进行定性、定量。结果:空白血、空白肝加标回收率在74%~120%,检出限为0.43 ng·mL^(-1)。结论:本文通过初步探讨维库溴铵大鼠体内分布,得知维库溴铵在大鼠体内的分布变化是受多种复杂因素影响的动态变化过程,并且死后随提取检材部位的不同测出药物浓度存在较大差异。真实案例中也证实了维库溴铵体内分布这一特点。本文可以为相关中毒案(事)件的检验提供方法,并且满足检验要求。

Objective： To establish a UPLC-MS/MS method for the quantitative determination of vecuronium bromide in whole blood and liver. Methods： The whole blood and blank liver vecuronium bromide and then diluted by ammonia water, followed by vortexing and centrifuging and confirmation were added with The supernatant liquid was cleaned-up by solid phase extraction technique using Oasis WCX. Extracts were analyzed by UPLC- MS/MS in positive ionization. An acquity UPLC~ BEH HILIC column was used with gradient elution of acetonitrile and5 mmol. L-1 ammonium formate buffer （ pH=3.5 ） as the mobile phase, and the signals were detected by ESI source in MRM mode operated in positive modes. The parent ion for vecuronium bromide is m/z 557.526. Two daughter ions were monitored at m/z 356. 346 and 100. 103 for quantification and confirmation. Results： The average recovery of vecuronium bromide from the whole blood samples and the blank liver was 74%-120%. The detection limit was 0. 43 ng. mL-1. Conclusion： From the preliminary discussion on distribution of vecuronium bromide in rats, it was known that the distribution variation of vecuronium bromide in rats was a dynamic variation process influenced by multiple complex factors and there was a postmortem maldistribution of vecuronium bromide in poisoning death rats. The distribution of VEC in poisoning death rats was confirmed by the real case. This paper could provide test method for related poisoning cases or events, and the method meets the test requirements.