【摘要】 目的建立及验证超高效液相色谱串联质谱(UPLC-MS,/MS)的方法用于测定人血浆和尿液中恶唑烷酮类新药MRX-I药物浓度。方法 UPLC-MS/MS液相条件为色谱柱Waters ACQUITY UPLC BEH C8;流动相为乙腈:水(40:60,v/v)。质谱采用ESI源正离子多反应监测(MRM)。内标为利奈唑胺,以乙酸乙酯液-液萃取法清除血浆及尿液样本中杂质。方法学验证包括基质效应、绝对回收率、精密度和准确度及MRX-I在人血浆及尿液样本中放置稳定性。结果 UPLC-MS/MS法检测MRX-I在人血浆和尿液中的线性范围均为(0.005 00~1.00)mg/L,最低检测浓度均为0.005 00 mg/L。MRX-I与内标在血浆和尿液中的保留时间小于1.5 min。本方法学验证结果显示MRX-I在人血浆和尿液基质效应因子分别为90.4%±8.2%和82.7%±7.9%;血浆和尿液中MRX-I提取回收率分别为112.8%±13.4%和105.6%±13.4%。MRX-I血浆样本的测定方法日内、日间准确度分别为98.9%~105.0%和96.5%~102.6%;尿液样本的测定方法日内、日间准确度分别为92.7%~98.6%和95.1%~105.7%。MRX-I在人血浆和尿液样本室温放置24 h、预处理后自动进样器放置48 h、-40℃冰箱冻融3次、-40℃冰箱分别放置8个月和6个月仍然保持稳定。结论本研究建立的UPLC-MS/MS检测人血浆及尿液中MRX-I浓度方法的灵敏度高,专属性强。其方法学验证结果均符合生物样品分析的要求。更多还原

【Abstract】 Objective To establish and validate an ultra performance liquid chromatography-tandem mass spectrometric(UPLCMS/MS) method for quantification of MRX-I,a new oxazolidinone antibacterial agent,in human plasma and urine.Methods Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C8 column using an isocratic elution.The mobile phase consisted of acetonitrile and water(40:60,v/v).Quantitative analysis was conducted in the multiple reaction monitoring mode.Linezolid was used as an internal standard.Liquid-liquid extraction with ethyl acetate was used to remove impurities in the plasma and urine samples.The method was validated in terms of matrix effect,recovery,precision,accuracy and stability.Results The calibration curves were linear within the range of 0.005 00-1.00 mg/L.The lower limit of quantification was 0.005 00 mg/L for both plasma and urine samples.Retention time was less than 1.5 min for both MRX-I and internal standard in plasma and urine.The matrix effect factors of plasma and urine for MRX-I was 90.4%± 8.2%and 82.7%±7.9%,respectively.The recovery of MRX-I was 112.8%± 13.4%from plasma and 105.6%±13.4%from urine samples,respectively.The inter- and intra-day accuracy of MRX-I was 98.9%-105.0%and 96.5%-102.6%in plasma samples,and 92.7%-98.6%and 95.1%-105.7%in urine samples.MRX-I was stable for 24 h at room temperature,48 h in automatic sampler after pretreatment,and stable after 3 freeze-thaw cycles in plasma and urine.MRX-I was also stable at-40℃ for eight months in plasma and six months in urine,respectively.Conclusions The UPLC-MS/MS method established in this study shows high sensitivity and specificity for determination of MRX-I in human plasma and urine.The results of validation are consistent with the requirement of bioanalytical method validation.更多还原