目的:建立人血浆中比阿培南的高效液相色谱检测方法.方法:色谱柱为Agilent ZORBAX Bonus-RP (4.6 mm×250 mm,5 μm);流动相为甲醇和0.03%醋酸水溶液,采用梯度洗脱法;流速为1.00 ml/min;检测波长为300 nm,血浆样品处理选用5-羟基吲哚乙酸为内标,采用硫酸锌沉淀蛋白.结果:比阿培南在0.2～50.0 μg/ml 范围内线性良好,HPLC 测定定量下限为0.2 μg/ml,比阿培南和内标在血浆的保留时间分别为3.9 min 和9.7 min,出峰迅速.比阿培南浓度分别为0.5、5.0 和40.0 μg/ml,血浆样本回收率分别为101.8%、100.8%、100.4%;精密度试验日内和日间RSD均小于15%,稳定性试验结果表明血浆样品在样品处理过程中较稳定(RSD<15%).结论:本方法灵敏度高,操作简便,快速,重复性好,适用于比阿培南血浆浓度的测定.%Objective: To establish a method for the determination of Biapenem in human plasma by HPLC. Methods: Agilent ZORBAX Bonus-RP (4.6 mmx250 mm, 5 |jLm) was used as the analytical column; mobile phase was composed of methanol-0.03% acetic acid solution (gradient elution); the flow rate was 1.00 ml/min; the LJV detection was carried out at 300 nm. The plasma samples were performed by protein precipitation with ZnSO4 using 5-hydroxyindole acetic acid as internal standard sample. Results: The assay was linear over the concentration range of 0.2-50.0 |JLg/ml for Biapenem in human plasma. The limit of quantitation for biapenem was 0.2 |jLg/ml. The retention time of Biapenem and internal standard sample in plasma was 3.9 min and 9.7 min, respectively. The recovery rate at concentrations of 0.5, 5.0, 40.0 |jLg/ml were 101.8%,100.8%,100.4%, respectively. The method was accurate with all intra-day and inter-day mean concentrations within 15% of nominal values. Furthermore, the stability results showed that Biapenem was stable during sample processing (RSD<15%). Conclusion: This method is high sensitive, operation simple, fast and repeatable, which is suitable for determination of Biapenem in human plasma.
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