HPLC-UV法检测人血浆中双氢可待因的浓度

Detection the Concentration of Dihydrocodeine in Human Plasma by HPLC-UV

目的建立人血浆双氢可待因检测的高效液相色谱方法。方法血浆经乙酸乙酯、正己烷萃取,以ZORBAXXDB-C18为色谱柱;流动相为乙腈-水-0.1%三氟乙酸(trifluoroacetic acid,TFA),流速为1.0 mL·min-1;检测波长为230 nm(0～5.0 min)和210 nm(5.0～6.2 min)。结果双氢可待因浓度在2.5～100.0μg·L-1范围内线性关系良好(r=0.9998);定量下限为2.5μg·L-1;高中低三个浓度的相对回收率分别为(101.48±5.87)%、(103.75±4.48)%和(100.06±2.51)%;日内相对标准偏差(Relative standard deviation,RSD)分别为5.79%、4.32%和2.51%,日间RSD分别为6.64%、4.05%和3.95%。结论本高效液相色谱紫外法准确可靠、简便快速,适用于人血浆双氢可待因浓度的测定及其药物代谢动力学研究。

Objective To develop a high performance liquid chromatography method for the determination of dihydroeodeine in human plasma. Methods The plasma was extracted by ethyl acetate and hexane. The analytical column was packed with ZORBAX XDB-C18. The mobile phase were acetonitrile-water -0. 1% trifluoroacetic acid and the flow rate was 1.0 mL · min^-1. The UV detection wavelength was 230 nm （0 -5.0 min） and 210 nm （5.0-6.2 min）. Results Excellent liner relationship was obtained from the range of 2.5 μg·L^-1 to 100.0μg·L^-1（ r = 0. 999 8 ）, the limit determination of dihydrocodeine was 2.5μg·L^-1. The relative recoveries were （ 101.48 ± 5.87 ） % , （ 103.75 ±4.48） % and（ 100.06 ± 2.51 ）% respectively at three concentrations, the intra-day RSD were 5.79% , 4.32% and 2. 51% and inter-day RSD were 6.64% , 4.05% and 3.95% respectively. Conclusion This chromatography method was accurate, simple and rapid and could be used to determine the dihydrocodeine concentration in human plasma and to study its pharmacokinetics.