摘要
A sensitive RP-HPLC-UV method has been developed and validated for the quantification of daphnoretin in rat plasma. Daphnoretin was extracted from rat plasma by protein precipitation and liquid-liquid extraction. Separation was performed on a Diamonsil C18 column （200 mm× 4.6 mm, 5 μm） with a mobile phase of methanol-20 mmol/L ammonium acetate （adjusted to pH 3.2 with acetic acid, 42：58, v/v） at a flow rate of 1.0 mL/min. The UV detector was set at 345 nm and column temperature was set at 40 ℃. The calibration curves were linear over the concentration range of 0.020-2.00 ~tg/mL, The lower limit of quantification （LLOQ） of daphnoretin in rat plasma was 0.020 μg/mL. The intra- and inter-day relative standard deviation （RSD） for measurement of quality control （QC） samples （0.050, 0.200 and 1.60 μg/mL） ranged from 5.0%-10.6%. Relative error （RE） was from ±（1.2%-2.5%）. The validated method was used successfully in a pharmacokinetic study of daphnoretin in rats after intraperitoneal injection.
建立了测定大鼠血浆中西瑞香素浓度的高效液相色谱法,并对其进行方法学确证。以蛋白沉淀法和液液萃取法对大鼠血浆中西瑞香素进行提取。采用Diamonsil C_(18)柱(200mm×4.6mm,5μm),流动相为甲醇-20mmol/L醋酸铵(醋酸调pH至3.2,42:58,v/v),流速1.0mL/min。检测波长345nm,柱温40℃。西瑞香素在0.020-2.00μg/mL浓度范围内线性关系良好(r>0.9900),定量下限0.020μg/mL。日内和日间精密度(RSD)为5.0%-10.6%,准确度(RE)为±(1.2%-2.5%)。所建立的方法成功应用于腹腔注射西瑞香素在大鼠体内的药代动力学研究。
基金
National Key Scientific Project for New Drug Discovery and Development(Grant No.2009ZX09301-012)
