高效液相色谱法测定人血浆中比阿培南的浓度

Determination of biapenem in human plasma by HPLC

目的建立人血浆中比阿培南的高效液相检测方法。方法色谱柱为Waters ACQUITY○RBEH C18柱(250×4.6mm,5μm),流动相为乙腈∶水(15∶85,v/v);流速1.0 mL.min-1;检测波长为300nm,血浆样品处理选用亚胺培南为内标,采用乙腈沉淀蛋白。结果比阿培南在0.05-25.0μg.mL-1范围内线性良好(r=0.9988,加权系数为1/C2),HPLC测定定量限为0.05μg.mL-1,比阿培南浓度分别为0.1,5.0 and 20.0μg.mL-1血浆样本绝对回收率分别为94.28%,95.07%,93.96%,相对回收率分别为99.87%,102.55%,98.64%,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测定。

OBJECTIVE To establish a HPLC method for the determination of biapenem in human plasma.METHODS The chromatography conditions were as follows： chromatographic column was Waters ACQUITY BEH C18 Column （250 × 4.6mm, 5μm）, mobile phase was composed of acetonitile-water （15 ： 85, v/v）, the flow rate was 1.0 mL·min^-1 and the UV detection was carried out at 300 nm. The plasma samples were performed by protein precipitation with acetonitrile using imipenem as internal standard. RESULTS The assay was linear over the concentration range of 0. 05-25. 0μg·mL^-1 for biapenem in human plasma （r = 0. 9988, weighted by 1/C^2）. The limit of quantitation for biapenem was 0.05μg·mL^-1. The absolute recoveries at concentrations 0.1,5.0 and 20.0μg·mL^-1 were 94.28 %, 95.07 %, 93.96 %, respectively, while the relative recoveries at concentrations 0.1,5.0 and 20.0μg·mL^-1 were 99.87 %, 102.55 %, 98.64 %, respectively. The method was accurate with all intra-and inter-day mean concentrations within 15 96 of nominal values at 0.1, 5.0 and 20.0μg·mL^-1. Furthermore, the stability results showed that biapenem was stable during sample processing （RSD 〈 15 % ） ）. CONCLUSION This method was robust and suitable for determination of biapenem in human plasma.