基于UHPLC-MS/MS法,中毒量氯化琥珀胆碱在大鼠体内分布的研究

Study on distribution of succinylcholine chloride at poisoning dose in rats by UHPLC-MS/MS analysis

目的建立适当的超高效液相色谱串联质谱(UHPLC-MS/MS)法,测定中毒量氯化琥珀胆碱(succinylcholine chloride,Suc)在大鼠体内的分布。方法大鼠分别皮下注射3种剂量的Suc,4 h后取大鼠血清和各主要脏器组织进行乙腈蛋白沉淀并过固相萃取WCX柱进行纯化;色谱采用Luna NH2色谱柱(2 mm×100 mm,3μm),流动相为0.1%甲酸水溶液-乙腈等度洗脱,质谱测定采用正离子扫描下多反应监测模式(MRM)。对Suc离子对(m/z 145.1→93.3,m/z 145.1→115.4)进行定性、定量分析。结果Suc的检测限是0.01μg·L^-1,回收率为89.5%~95.4%,日内、日间RSD均小于15%。除NS对照组外,3种剂量的Suc组大鼠的血清及心、肝、肾等主要脏器组织中均检测到Suc,并且肾组织中的Suc最高。结论大鼠Suc中毒时在血清及心、肝、肾等组织中均有分布,其中肾脏中含量最高;UHPLC-MS/MS法是检测Suc中毒大鼠体内药量的一个有效、可靠的方法。

Aim To determine the distribution of suc-cinylcholine chloride(Suc)at toxic dose in rats byestablishing UHPLC-MS/MS analysis.Methods The rats were subcutaneously injected with three doses of Suc,and 4 h later,rat serum and major organs were taken for acetonitrile protein precipitation and purified by solid phase extraction with WCX column.Luna NH2 column(2 mm×100 mm,3μm)was used for chromatography.The mobile phase was 0.1%formic acid-acetonitrile isoelution.Mass spectrometry was carried outwith positive ion scanning multi-reaction monitoring mode(MRM).Qualitative and quantitative analysis was conducted on the ion pairs of Suc(m/z 145.1→93.3,m/z 145.1→115.4).Results The detection limit of Suc was 0.01μg·L^-1,the recovery rate was 89.5%~95.4%,and the intraday and intraday RSD was less than 15%.Except in the untreated rat,Suc was detected in the serum,heart,liver,kidney and other major organs from Suc-administered rats,and Suc content in the kidney was the highest among organs.Conclusions Sucat poisoning dose is distributed in the serum,heart,liver,kidney and other tissues.The content in the kidney is the highest;UHPLC-MS/MS analysis is an effective and reliable method for detecting the drug content in Suc-poisoned rats.