LC-MS/MS法测定比格犬血浆中的磷酸可待因

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

目的建立灵敏、快速的LC-MS/MS法测定比格犬血浆中磷酸可待因浓度的方法。方法血浆样品经乙醚-二氯甲烷(60∶40)萃取后,以甲酸-10 mmol·L-1乙酸-甲醇(0.2∶62∶38)为流动相,盐酸利多卡因为内标,采用Phenomenex C18(150 mm×2 mm,3μm)色谱柱进行分离,流速0.2 mL·min-1,进样量10μL;LC-MS/MS法选择反应监测方式,定量分析的离子反应分别为m/z 300→165(磷酸可待因)和m/z 235→86(盐酸利多卡因)。结果 0.08～16 ng·mL-1磷酸可待因与峰面积的线性关系良好(r=0.9977);其最低定量下限为0.08 ng·mL-1,绝对回收率大于85%,批间和批内的变异系数小于15%。结论所用方法操作简便、快速、选择性好、灵敏度高,适用于磷酸可待因的生物等效性及药物动力学的研究。

OBJECTIVE To establish a rapid and sensitive method based on LC-MS/MS to determine the content of codeine in Beagle dogs plasma.METHODS After being extracted by diethyl ether-dichloromethane（60：40）,the plasma samples were separated on Phenomenex C18 column（150 mm×2 mm,3 μm） using formic acid-10 mmol·L-1 ammonium acetate-methanol（0.2：62：38） as mobile phase and Lidocaine Hydrochloride as internal standard.The flow rate was 0.2 mL·min-1,and the volume of injection was 10 μL.The detection was performed on a HPLC-triple quadrupole tandem mass spectrometer by selected reaction monitoring（SRM） mode via electrospray ionization（ESI）.In quantitative analysis,the ionic reaction of codeine was m/z 300→165 and that of lidocaine was m/z 235→86.RESULTS The method was linear in the concentration range of 0.08-16 ng·mL-1（r=0.9977） for codeine,with lower limit of quantification of 0.08 ng ·mL-1,and the absolute recovery was greater than 85%.The within-run and between-run relative standard deviation was less than 15%.CONCLUSION The established method is simple,rapid,sensitive,and suitable for pharmacokinetic study of codeine.