HPLC法同时测定健康人血浆中贝那普利及其代谢物贝那普利拉的浓度

Determination of benazepril and benazeprilat in healthy human plasma by high-performance liquid chromatography

目的建立一种灵敏度较高的高效液相色谱法,可同时测定健康人血浆中贝那普利及其代谢物贝那普利拉的浓度。方法流动相用0.02mol·L-1磷酸二氢钾缓冲液-乙腈(7∶3)(pH2.6),用取梯度洗脱法,血浆样本用C8固相小柱萃取,紫外检测波长为237nm。结果贝那普利:在0.06～2μg·mL-1,线性关系良好;最低检测限为0.03μg·mL-1;日内RSD≤6.3%,日间RSD≤4.0%;提取回收率在73%～78.5%(n=5),相对回收率在93.2%～101.3%。贝那普利拉:在0.06～2μg·mL-1,线性关系良好;最低检测限为0.03μg·mL-1;日内RSD≤8.5%,日间RSD≤6.0%;提取回收率在67.7%～74.1%(n=5),相对回收率在95.6%～103.2%。结论本方法灵敏度高、重现性好,可用于贝那普利人体生物利用度和药代动力学研究。

Objective To develop a HPLC method for quantifying benazepril and benazeprilat in healthy human plasma. Methods Plasma samples were cleaned up by extraction with Cs solid- phase column. The chromatogram was monitored by UV detection at a wavelength of 237 nm. The mobile phase consisted of patassium dihydrogen phosphate - acetonitril pH 2. 6 （7：3）. The flow rate is gradient. Results A linear range of benazepril was 0.06 - 2 μg · mL^- 1 , the lower limit of detection of benazepril was 0.03 μg · mL^-1. The intra- and inter- run precision were lower than 6.3% and 4.0% （n = 5） , respectively. The extraction recovery was 73% - 78.5% （n = 5） and the method recovery was 93.2% - 101.3%. The linear range of benazeprilat was 0.06 - 2 μg · mL^-1, the lower limit of detection of benazeprilat was 0.03 μg · mL^-1 The intra- and inter- run precision were lower than 8.5% and 6.0% （n =5） , respectively. The extraction recovery were 67.7% - 74.1% （ n = 5 ） and the method recoveries were 95.6% - 103.2%. Conclusion The method is shown to be accurate and repetitive, simple to operate, suitable for pharmacokinetic studies of benazepril in human.