LC-MS/MS法测定Beagle犬血浆中可乐定的浓度

Determination of donidine in Beagle dogs plasma by LC-MS/MS

目的建立一种LC-MS/MS法,测定Beagle犬血浆中可乐定的质量浓度,从而对盐酸可乐定缓释片在犬体内的药物动力学行为进行研究。方法采用液-液萃取法制备血浆样品;色谱柱:Luna C_(18)柱,采用梯度洗脱方式,流动相:A为甲醇(含体积分数为0.3%的甲酸溶液),B为5 mmol·L^(-1)醋酸铵缓冲溶液(含体积分数为0.3%的甲酸溶液);采用多反应监测扫描方式,测定Beagle犬口服给予盐酸可乐定缓释片后血浆中可乐定的质量浓度。结果 Beagle犬血浆中可乐定的质量浓度在25~1 000 ng·L^(-1)内线性关系良好,相关系数r=0.998 6;日内和日间精密度不大于13.2%;犬血浆样品中可乐定的提取回收率在76.0%~82.3%内,基质效应在100.3%~112.0%内。结论该方法适合于犬体内可乐定的药物动力学的研究。

Objective To develop a liquid chromatography-tandem mass spectrometry（LC-MS/MS） method for the determination of clonidine in Beagle dogs plasma,and to study the pharmacokinetic characteristic of clonidine in Beagle dogs plasma.Methods Plasma samples were pretreated by liquid-liquid extraction with ethyl acetate.The chromatographic separation was accomplished on Luna C_（18） analytical column using gradient elution of methanol（solvent A） and 5 mmol·L^-1 ammonium acetate solution（solvent B） containing0.3%formic acid,respectively.The plasma concentration of clonidine was detected by multiple reaction monitoring after the Beagle dogs were given clonidine ex tended-release.Results The method was linear in the concentration range of 25.0-1 000 ng·L^-1 with a correlation coefficient of 0.998 6;the intra-and inter-day precision was both less than 13.2%.The average recoveries were between 76.0%and 82.3%.The matrix effects were between 100.3%and 112.0%.Conclusions This LC-MS/MS method is successfully applied to pharmacokinetic studies of clonidine in Beagle dogs plasma.