高效毛细管电泳测定血清中盐酸反式曲马多对映体

Determination of the Enantiomers of Trans Tramadol Hydrochloride in Human Serum by High Performance Capillary Electrophoresis

目的 :建立测定人血清中盐酸反式曲马多对映体的高效毛细管电泳方法。方法 :血清中盐酸曲马多以乙酸乙酯提取、浓缩。含 0 8mmol·L-1磺丁基 - β -环糊精的 40mmol·L-1Tris缓冲液 (磷酸调pH至 2 5 )为分离介质 ;进样电压 10kV ,2 0s,入口为阳极 ;分离电压 15kV ,柱温 2 5℃ ,检测波长 2 14nm。结果 :盐酸曲马多的 4种立体异构体达到基线分离 ,且血清组分无干扰峰 ;以盐酸顺式曲马多的一对映体为内标 ,(+) -盐酸反式曲马多、 (- ) -盐酸反式曲马多的血清浓度在 2 5～ 32 0ng·mL-1范围内 ,峰面积与内标峰面积的比值和浓度呈良好的线性关系 ;回收率在 90 %和 10 8%之间 ;日内、日间RSD分别小于 18%和 2 1% ;最低检测浓度为1 2 5ng·mL-1。结论 :本方法适用于临床药代动力学研究。

Objective:To establish a high performance capillary electrophoresis(HPCE)method for determining the two enantiomers of trans tramadol hydrochloride in human serum.Methods:The tramadol hydrochloride in human serum was extracted by ethyl acetate and concentrated.The electrophoretic medium was 40 mmol·L -1 Tris buffer(pH adjusted to 2 5 by H 3PO 4)containing sulfobutylether-β-cyclodextrin.An uncoated capillary used was 37 cm(30 cm to the detector)×75 μm.Detection was carried out with a UV detector at 214 nm.The separations were performed at 25 ℃ with a positive voltage of 15 kV.Samples were injected into the capillary by voltage (10 kV) for 20 s.Results:With one enantiomer of cis tramadol hydrochloride as internal standard,the two enantiomers of trans tramadol hydrochloride were separated well and without interference from human serum.The linear concentration region was 2 5～320 ng·mL -1 for each enantiomer.The relative recoveries were from 90% to 108%.The within-day and between-day RSD s were lower than 18% and 21% respectively.The lowest detective concentration was 1 25 ng·mL -1 for each enantiomer. Conclusion:This method is very suitable for clinical pharmacokinetic study.