HPLC法同时测定人血浆中氢氯噻嗪与奥美沙坦浓度

Simultaneous determination of hydrochlorothiazide and olmesartan in human plasma by HPLC

目的建立同时测定人血浆中氢氯噻嗪、奥美沙坦浓度的HPLC法。方法采用Kromasil C18柱，以乙腈-0．02mol·L^-1 KH2PO4（32：68，V/V；磷酸调pH2．40）为流动相，坎地沙坦为内标，检测波长为272nm，血浆样品采用盐酸酸化后用EO基叔丁基醚提取浓集，按内标法定量。结果氢氯噻嗪血药浓度在0．005—0．64mg·L^-1内、奥美沙坦血药浓度在0．01～4．0mg·L^-1内线性关系良好（r=0．9991、r=0．9988）。氢氯噻嗪日内精密度（RSD）为3．11％～7．29％，日间RSD为3．72％～10．54％；奥美沙坦日内RSD为0．97％～5．18％，日间RSD为5．37％～9．17％。氢氯噻嗪提取回收率为78．38％～80．77％，相对回收率为98．01％．101．26％；奥美沙坦提取回收率为72．28％～74．35％，相对回收率为97．04％～100．17％。结论本法简便、灵敏、准确、重现性好，可用于人血浆中氢氯噻嗪、奥美沙坦浓度的测定。

AIM To establish a method for simultaneously determining the concentration of hydrochlorothiazide and olmesartan in human plasma by HPLC. METHODS The analysis involved a Kromasil C18 column and the mobile phase was constituted of aeetonitrile-0.02 mol· L^-1 potassittm dihydrogen phosphate buffer （32：68, V/V;The pH was adjusted to 2.40 with phosphoric acid）. The detective wavelength was set at 272 nm and candesartan was used as the in- ternal standard. Plasma samples of hydrochlorothiazide and olmesartan were extracted with methyl tert-butyl ether after acidification with hydrochloric acid. RESULTS Calibration curves of hydrochlorothiazide and olmesartan were linear in the concentration range of 0.005 to 0.64 mg· L^-1 （ r = 0.999 1 ） and 0.01 to 4.0 mg· L^-1 （ r = 0.998 8）, respectively. The intra-day RSD and inter-day RSD of hydrochlorothiazide vs olmesartan were 3.11% -7.29% vs 0.97% -5.18% and 3.72% - 10.54% vs 5.37 % - 9.17 %, respectively. The absolute recovery and relative recovery of hydrochlorothiazide vs olmesartan were 78.38% -80.77% vs 72.28% -74.35% and 98.01% - 101.26% vs 97.04% 100.17%, respectively. CONCLUSION The method is found to be simple, sensitive, accurate with good reproducity for determining the concentration of hydrochlorothiazide and olmesartan in human plasma.