LC-MS/MS法测定大鼠尿液和粪便中WSJ-537的浓度及其在大鼠体内的排泄研究

Determination of WSJ-537 concentration in rat urine and feces by LC-MS/MS and its excretion in rats

目的建立LC-MS/MS法测定大鼠尿液和粪便中WSJ-537的浓度,并研究其在大鼠体内的排泄行为。方法使用乙腈沉淀法进行排泄物样品预处理,选用乙氧苯柳胺作为内标进行检测。色谱柱:AcquityTM UPLC BEH C18柱(2.1 mm×50 mm,1.7μm);流动相:水(含0.1%甲酸)-乙腈(含0.1%甲酸),梯度洗脱;流速:0.3 mL·min^-1。LC-MS/MS法测定大鼠尿液和粪便中WSJ-537的浓度。结果WSJ-537在4~4000 ng·mL-1与峰面积线性关系良好,所建立方法的准确度、精密度、提取回收率、基质效应和稳定性均符合要求。WSJ-537单剂量灌胃给药5.2 mg·kg^-1后,大鼠尿液中的累积排泄总量为(235.09±316.88)ng,大鼠粪便中的累积排泄总量为(3742.74±2267.77)ng,排出体外的原形药物总量约占总给药量的0.36%。结论该方法预处理步骤简便,分析时间短,线性范围宽,灵敏度高,可用于研究WSJ-537在大鼠体内的排泄行为。

Objective To determine the concentration of WSJ-537 in the urine and feces of rats by LC-MS/MS,and study the excretion behaviors in vivo.Methods The excretion sample was precipitated with acetonitrile.Elofesalamide was used as the internal standard.An AcquityTM UPLC BEH C18 column(2.1 mm×50 mm,1.7μm)was used.The mobile phase was 0.1%formic acid-water and 0.1%formic acid-acetonitrile with gradient elution.The flow rate was 0.3 mL·min^-1.The concentration of WSJ-537 in the urine,feces of rats was determined.Results The linearity of WSJ-537 was good at 4~4000 ng·mL^-1.The accuracy,precision,extraction recovery,matrix effect and stability of the established method all met the requirements.After a single dose oral administration of WSJ-537 at 5.2 mg·kg^-1,the cumulative excretion in the urine in rats was(235.09±316.88)ng,and the cumulative excretion in the feces in rats was(3742.74±2267.77)ng.The proportion of WSJ-537 excreted in vitro in the form of prototype compounds accounted for about 0.36%of the total administration.Conclusion The method has simple pretreatment procedure,short analysis time,wide linear range,and high sensitivity,and can be used to study the excretion behaviors of WSJ-537 in rats.