高效液相色谱法测定盐酸替络肟血药浓度

Determination of tiloronoxim hydrochloride in human blood by HPLC

目的:建立一种同时测定人全血中盐酸替络肟及其代谢产物替络龙的高效液相色谱法。方法:色谱柱为YMC-Pack ODS-A柱(150 mm×6．0 mm,5μm),以0．2 mol·L-1磷酸盐缓冲液(pH=4．6)-甲醇(58:42)作为流动相,酒石酸美托洛尔作为内标物,流速1．0 mL·min-1,紫外检测波长为263 nm。采用液液萃取方法同时测定人全血中盐酸替络肟及其代谢产物盐酸替络龙的浓度。结果:盐酸替络肟血药浓度线性范围为50-5 000 ng·mL-1,低、中、高(125,625,2 500 ng·mL-1)3个血药浓度的日内RSD分别为2．6％,2．0％和0．1％,日间RSD分别为6．2％,2．3％和0．3％,方法准确度分别为(99．9±0．01)％,(101．0±0．04)％和(100．0±0．01)％。盐酸替络龙血药浓度线性范围为50-5 000 ng·mL-1,低、中、高(125,625,2 500 ng·mL-1)3个血药浓度的日内RSD分别为5．1％,3．1％和1．1％,日间RSD分别为5．8％,1．3％和0．7％。方法准确度分别为(101．0±0．03)％,(97．8±0．03)％和(99．2±0．02)％。结论:建立的HPLC法可以同时测定人全血中盐酸替络肟及其代谢物的血药浓度,方法操作简便、准确、专一,满足该药临床药动学研究的需要。

Objective：To establish a HPLC method for determination of tiloronoxim hydrochloride and its metabolite （tilorone hydrochloride） in human blood. Methods： The basified plasma samples from volunteers who administered tiloronoxim were extracted with chloroform-ether （ 1 ： 2 ） and measured by a HPLC consisted of a mobile phase containing phosphate buffer（ pH 4. 6）-methanol （58：42） and a detective wavelength at 263 nm. The flow rate was I. 0 mL.min^-1 Results ： The calibrated linear curve of tiloronoxim concentration was in the range of 50 -5 000 ng.mL^-1 （r =0. 999 1 ）. The samples at 125,625 and 2 500 ng.mL-1 showed within day （n =5） precisions （RSD） of 2.6% , 2.0% and 0. I% , respectively, between day （n =5） precisions （RSD） of 6.2% , 2. 3% and 0.3% , respectively, and an average recovery of （99.9 ＋0. 01 ）% , （ 101.0 ＋0. 04）% and （ 100.0 ＋0. 01 ）% , respectively. The calibrated linear curve of tilorone concentration was in the range of 50 - 5 000 ng. mL^- 1 （r = 0. 999 1 ）. The metabolite samples at 125,625 and 2 500 ng. mL^-1 gave within day （ n = 5 ） precisions （RSD） of 5. 1% , 3. 1% and 1.1%, respectively, between day （n=5） precisions （RSD） of 5.8%, 1.3% and 0.7%, respectively, and an average recovery of （ 101.0 ＋ 0.03 ） % , （97.8 ＋ 0. 03 ） % and （99.2 ＋ 0.02） % , respectively. Conclusion： This sensitive, reliable and easy analytical method may be applied in pharmacokinetic studies of tiloronoxim hydrochloride and its metabolite.