建立了以阿莫西林d4为稳定同位素标记内标的液相色谱-电喷雾串联质谱同时快速检测人血浆中阿莫西林和克拉维酸的方法,并进行了验证。在HederaODS-2色谱柱上进行色谱分离(2.1×150mm,5μm)。梯度洗脱的流动相为含0.2%乙酸(AA)的水溶液(流动相A)和有机相溶液(乙腈和甲醇混合溶液,流动相B)。在多反应监测模式下使用负电喷雾电离进行质谱。阿莫西林的靶碎片离子对,克拉维酸和阿莫西林-d4分别为m/z364.1→223.1、198.1→135.9和368.1→227.1。该方法的线性范围为阿莫西林40-5,000ng/ml,克拉维酸30-2,500ng/ml,决定系数>0.9900。该方法验证包括选择性,标准曲线,定量下限,准确度,精度,recovery,基质效应(溶血基质和高脂血症基质),结转,稳定性,稀释可靠性和招致样品再分析研究。在服用阿莫西林-克拉维酸钾颗粒后,在药代动力学研究中成功应用了该方法。
A liquid chromatography electrospray ionization tandem mass spectrometry method with amoxicillin-d4 as the stable isotope-labeled internal standard for simultaneous quick detection of amoxicillin and clavulanic acid in human plasma was developed and validated. Chromatographic separations were performed on a Hedera ODS-2 column (2.1 × 150 mm, 5 μm). The mobile phases for gradient elution were aqueous solution containing 0.2% acetic acid (AA) (mobile phase A) together with organic phase solution (acetonitrile and methanol mixed solution, mobile phase B). Mass spectrometry was performed using negative electrospray ionization in multiple reaction monitoring mode. The target fragment ion pairs of amoxicillin, clavulanic acid and amoxicillin-d4 were m/z 364.1 → 223.1, 198.1 → 135.9 and 368.1 → 227.1, respectively. The linear ranges of this method were 40-5,000 ng/ml for amoxicillin and 30-2,500 ng/ml for clavulanic acid, with coefficient of determination > 0.9900. This method validation included selectivity, standard curve, lower limit of quantitation, accuracy, precision, recovery, matrix effect (hemolytic matrix and hyperlipidemic matrix), carryover, stability, dilution reliability and incurred sample reanalysis study. A successful application of this method was realized in a pharmacokinetic study after administration of amoxicillin-clavulanic acid potassium granules.