CAJ | 학술논문

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口恶唑烷酮类新药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（0．00500～1．00）mg/L，最低检测浓度均为0．00500 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。
목적：건립급험증초고효액상색보천련질보（UPLC-MS/MS）적방법용우측정인혈장화뇨액중구악서완동류신약MRX-I 약물농도。방법UPLC-MS/MS 액상조건위색보주 Waters ACQUITY UPLC BEH C8；류동상위을정∶수（40∶60， v/v）。질보채용 ESI 원정리자다반응감측（MRM）。내표위리내서알，이을산을지액-액췌취법청제혈장급뇨액양본중잡질。방법학험증포괄기질효응、절대회수솔、정밀도화준학도급 MRX-I 재인혈장급뇨액양본중방치은정성。결과UPLC-MS/MS법검측 MRX-I 재인혈장화뇨액중적선성범위균위（0．00500～1．00）mg/L，최저검측농도균위0．00500 mg/L。MRX-I 여내표재혈장화뇨액중적보류시간소우1．5 min。본방법학험증결과현시 MRX-I 재인혈장화뇨액기질효응인자분별위90．4％±8．2％화82．7％±7．9％；혈장화뇨액중 MRX-I 제취회수솔분별위112．8％±13．4％화105．6％±13．4％。MRX-I 혈장양본적측정방법일내、일간준학도분별위98．9％～105．0％화96．5％～102．6％；뇨액양본적측정방법일내、일간준학도분별위92．7％～98．6％화95．1％～105．7％。MRX-I 재인혈장화뇨액양본실온방치24 h、예처리후자동진양기방치48 h、-40℃빙상동융3차、-40℃빙상분별방치8개월화6개월잉연보지은정。결론본연구건립적 UPLC-MS/MS 검측인혈장급뇨액중MRX-I 농도방법적령민도고，전속성강。기방법학험증결과균부합생물양품분석적요구。
Objective To establish and validate an ultra performance liquid chromatography-tandem mass spectrometric (UPLC-MS/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 mo-bile phase consisted of acetonitrile and water (40∶60,v/v).Quantitative analysis was conducted in the multiple reaction moni-toring mode.Linezolid was used as an internal standard.Liquid-liquid extraction with ethyl acetate was used to remove impuri-ties 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 ma-trix 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 tem-perature,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 re-sults of validation are consistent with the requirement of bioanalytical method validation.