UPLC-MS/MS法测定犬血浆中的盐酸右哌甲酯

目的建立犬血浆中盐酸右哌甲酯的UPLC-MS/MS检测方法。方法用甲基叔丁基醚提取犬血浆,离心后取上清液用氮气吹干,用甲醇-水(80∶20)溶解后进行质谱分析。采用BEH C18色谱柱(50 mm×2.1 mm,1.7μm),流动相为甲醇-0.2%甲酸(80∶20),流速0.1 mL·min-1,柱温45℃,进样量5μL。电喷雾离子化(ESI),正离子模式多重反应选择离子检测(MRM),盐酸右哌甲酯及卡马西平内标的检测离子对分别为m/z 234.33→84.56和237.35→194.31。结果盐酸右哌甲酯的线性范围为0.1～50 ng·mL-1(r=0.994,n=5),在犬血浆基质中,高、中、低浓度的日内、日间RSD均小于15%,方法的回收率为89%～103%,血浆基质对测定无干扰。结论所用方法处理简单、灵敏、特异性高,定量准确,为盐酸右哌甲酯制剂的临床药物动力学研究提供了简便、准确的分析测定方法。

Determination of (+) -threo-methylphenidate hydrochloride in the dog plasma by UPLC-MS/MS

OBJECTIVE To establish a method for determination of（+）-threo-methylphenidate hydrochloride in dog plasma by UPLC-MS/MS.METHODS Plasma sample was extracted by methyl tert-butyl ether.The resulting supernatant liquid after centrifuge was dried by nitrogen flow and analyzed by UPLC-MS/MS method after diluted quantitatively by methanol-water（80:20）.The following chromatography conditions were used:BEH C₁8（50 mm×2.1 mm,1.7 μm）,the mobile phase consisted of methanol-0.2% formic acid（80:20）,the flow rate was 0.1 mL·min^-1,column temperature was 45 ℃,and injection volume was 5 μL.Positive ion MRM detection of（+）-threo-methylphenidate hydrochloride（m/z 234.33→84.56）and carbamazepine（m/z 237.35→194.31）was used as internal standard.RESULTS A good linearity was shown in the range of 0.1-50 ng·mL^-1（r=0.994,n=5）.Intra-day RSD and inter-day RSD were less than 15%.The recovery was within the range of 89%-103%.Plasma matrix effect test showed endogenous matrix had no effect on quantification of（+）-threo-methylphenidate hydrochloride.CONCLUSION A rapid,sensitive and accuracy UPLC-MS/MS method was developed,validated for the determination of（+）-threo-methylphenidate hydrochloride in dog plasma.