摘要
A new HPLC-UV method was developed and validated for the quantitative determination of epidermal growth factor receptor inhibitor erlotinib in the plasma of tumor bearing BALB/c nude mice.Erlotinib and its internal standard l-（3-（（6,7-bis（2- methoxyethoxy）quinazolin-4-yl）amino）phenyl）ethanone were extracted from mice plasma samples using liquid-liquid extraction with a mixed solvent of methyl t-butyl ether and ethyl acetate（9：1,v/v）.Chromatographic separation was performed on a Luna C｜_（18）column（4.6 mm×250 mm,5μm）with acetonitrile：5 mM potassium phosphate buffer pH=5.2（41：59,v/v）as the mobile phase.UV detector was set at the wavelength of 345 nm,and the flow rate was 1.0 mL/min.The calibration curve was linear over the range of 20-10 000 ng/mL with acceptable intra-and inter-day precision and accuracy.The intra-day and inter-day precisions were within the range of 1.69%—5.66%,and the accuracies of intra-and inter-day assays were within the range of 105%—113%. The mean recoveries were 85.2%and 96.1%for erlotinib and IS,respectively.This method was successfully applied to a pharmacokinetic study in tumor bearing BALB/c nude mice following single oral administration at the dose of 12.5 mg/kg. The main pharmacokinetic parameters were as follows：C_（max）was 4.67μg/mL,T_（max）was 1.0 h,T_（1/2）was 2.78 h,and AUC_（0-24h）was 18.0μg/mL·h.
本试验建立并验证了测定肿瘤模型BALB/c裸鼠血浆中表皮生长因子受体抑制剂埃罗替尼血药浓度的高效液相色谱方法。使用液液萃取的方法,用甲基叔丁醚与乙酸乙酯(9:1,v/v)的混合溶剂将血浆中埃罗替尼和内标3-(6,7-双(2-甲氧基乙基)喹唑啉-4-氨基)苯乙酮提取。使用Luna C_(18)(4.6 mm×250 mm,5μm)分析柱,流动相为乙腈-5 mM的KH_2PO_4缓冲盐(41:59,v/v),pH为5.2,紫外检测波长为345 nm,流速为1.0 mL/min。本方法在20-10 000 ng/mL的浓度范围内具有良好的线性关系,并且具有可接受的日内和日间精密度和准确度,精密度在1.69%-5.66%的范围内,准确度在105%-113%的范围内。埃罗替尼和内标的平均提取回收率分别为85.3%和96.1%。该方法成功地运用到肿瘤模型BALB/c裸鼠单剂量口服给药12.5 mg/kg埃罗替尼后的药代动力学研究,主要的药动学参数如下:C_(max)为4.67μg/mL,T_(max)为1.0 h,T_(1/2)为2.78 h,AUC_(0-24)h为18.0μg/mL·h。
基金
National Integrity Innovational Technology Platform of New Drug and Development(Grant No.2009ZX09301- 010).
