反相高效液相色谱法测定曲马多及其代谢产物O-去甲基曲马多人血药及尿药浓度

Determination of tramadol and its metabolite O-demethyltramadol concentration in human plasma and urine by RP-HPLC

目的建立血浆和尿液中曲马多(镇痛药)及其代谢产物O-去甲基曲马多的反相高效液相色谱分析方法。方法用zorbax SB-C18色谱柱,0.05mol.L-1KH2PO4(用磷酸调节pH至4.0)-乙腈(90:10)为流动相,流量1.0mL.min-1;荧光检测波长Ex=216nm,Em=308nm。结果血浆中,曲马多、O-去甲基曲马多的线性范围分别为12.5～800.0、5～320ng.mL-1,低、中、高3种浓度绝对回收率均大于86%,相对回收率为93%～105%,日内和日间RSD分别小于9%和8%。尿液中,曲马多、O-去甲基曲马多线性范围分别为12.5～4000、5～1280ng.mL-1,低、中、高3种浓度绝对回收率均大于80%,相对回收率为91%～107%,日内和日间RSD均小于9%。在血浆和尿液中均稳定,差异均在±10%以内。结论本方法准确可靠、专一性好、操作简便。

Objective To establish analytical method for the determination of tramadol and its metabolite O - demethyhramadol in human plasma and urine by RP- HPLC. Methods Separation was carried out ons a zorbax RP - select B column and the mobile phase was a mixture of 0. 05 mol · L^-1 potassium dihydrogen phosphate （adjusted to pH 4. 0 with phosphoric acid）-acetonitrile （90： 10）. The flow rate was 1 mL ·min^-1 Fluormetric detection was performed at excitation and emission wavelengths of 216 nm and 308 nm, respectively. Results In plasma, the calibration curve was linear within 12.5 - 800. 0 ng · mL^- 1 for tramadol and 5 -320 ng · mL^-1 for O - demethyhramadol. The absolute recoveries were all higher than 86% , and the relative recoveries were from 93% to 105%. The RSD of the within - day and between - day variations were lower than 9% and 8% , respectively. In urine, the calibration curve was linear within 12. 5 -4 000 ng · mL^- 1 for tramadol and 5 - 1 280 ng · mL^-1 for O- demethyhramadol. The absolute recoveries were all higher than 80%, and the relative recoveries were from 91% to 107%. The RSD of the within-day and between-day variations were lower than 9%. For stability test in plasma and urine, the difference of the results was within 10%. Conclusion This RP- HPLC method is simple, sensitive, accurate and specific.