摘要
建立一种快速、准确测定人血浆中罗红霉素浓度的UPLC-MS/MS分析方法。以克拉霉素为内标,0.2mL含药血浆经碱化、乙酸乙酯萃取后进样分析;色谱柱为Acquity UPLC BEH C18（2.1mm×50mm×1.7μm）,流动相组成为乙腈∶0.01%醋酸铵=30∶70,梯度洗脱方式,乙腈比例在4 min内从30%变为70%,流速0.3 mL/min,柱温为35℃,进样量3μL。质谱条件：气动辅助电喷雾离子化（ESI）源;正离子检测（MRM）模式,罗红霉素质荷比为（m/z 837.53→m/z 158.15）和克拉霉素质荷比为（m/z 748.48→m/z 590.30）。罗红霉素在0.05~25.6μg/mL的浓度范围内呈线性,定量下限为0.05μg/mL,基质效应影响小,日内变异系数小于10.3%,日间变异系数小于9.4%,相对回收率在97.5%~106.4%之间。该方法准确、快速、灵敏,可用于微量血浆的罗红霉素药物浓度监测、人体内药代动力学及生物等效性研究。
To establish a rapid and accurate method for quantification of roxithromycin in human plasma with UPLC-MS/MS, human plasma sample was alkalized by sodium carbonate, extracted with ethyl acetate, the roxithromycin and internal standard clarithromycine were separated on an acquity UPLC BEH C18 column（2.1 mm×50 mm× 1.7 μm） using gradient elution with acetonitrile（mobile phase A） and 0.01% ammonium acetate（mobile phase B） with a flow rate of 0.3mL/min and a total runtime of 4.0 min. Mass spectrometry： The analytes were detected by the electronicspryionization（ESI）,selected multiple reaction monitoring（MRM） using the precursor to product ion combinations of m/z 837.53 →158.15 and m/z 748.48 →590.30 was performed to detect roxithromycin and the internal standard respectively. The method was linear over the concentration range of 0.05-25.6 μg/mL. The lower limit of quantification was 0.05 μg/mL. Intra-day inaccuracy was within 10.3% and inter-day was less than 9.4%. Matrix effects were minor. The recovery of the method was between 97.5% and 106.4%. The validated method was developed for quantification of roxithromycin in small volumes of plasma. The method can be used for therapeutic drug monitoring and the study of pharmacokinetics and bioavailability of roxithromycin.
出处
《中国测试》
CAS
北大核心
2013年第6期60-63,103,共5页
China Measurement & Test
