LC-MS/MS测定小鼠血浆中厄他培南的浓度
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摘要
目的建立一种快捷、简便、灵敏的LC-MS/MS方法测定小鼠血浆中的厄他培南。方法以XZP-3408为内标,血浆样品经过甲醇沉淀蛋白,离心后取上清液进行分析。色谱柱:ZORBAX Eclipse plus C18(4. 6 mm×50. 0 mm,3. 5micron);流动相:乙腈含0. 2%甲酸-0. 2%甲酸水溶液含10 mmol·L~(-1)乙酸铵;梯度洗脱;流速:0. 4 m L·min~(-1);进样量:5μL。电喷雾正离子检测方式,多反应监测。考察该方法的专属性、标准曲线与定量下限、精密度与回收率、基质效应、残留效应、稀释效应和稳定性。结果厄他培南在8～2000 ng·m L~(-1)内,线性关系良好,定量下限为8 ng·m L~(-1),批内及批间精密度分别为≤4. 20%和≤5. 81%,样品及内标回收率分别为96. 22%～100. 40%和88. 29%～96. 50%,日内及日间准确度均为-9. 30%～14. 00%,样品及内标基质效应分别为86. 10%～94. 87%和90. 01%～97. 32%,残留效应、稀释效应和稳定性等方法学考察的结果均符合要求。结论本文建立了一种灵敏、快速、稳定的LC-MS/MS方法用于测定小鼠血浆中厄他培南的浓度。
Objective To establish a rapid,sensitive method for the determination of ertapenem in mice plasma. Methods XZP-3408 was taken as the internal standard,the plasma samples were extracted by methanol to precipitate protein. After centrifugation,the supernatant was analyzed by LC-MS/MS. Chromatographic column of ZORBAX Eclipse plus C18( 4. 6 mm × 50. 0 mm,3. 5 micron),mobile phase composed of acetonitrile containing 0. 2% formic acid-water( 0. 2% formic acid)containing 10 mmol ·L~(-1) ammonium acetate in gradient elution mode,flow-rate of 0. 4 m L·min~(-1),injection volume of 5 μL. Electrospray positive ion detection method,multi-reaction monitoring. The specificity,standard curve and lower limit of quantification,precision and recovery,matrix effect,residual effect,dilution effect and stability of the method were investigated. Results Calibration curves were linear between 8 and 2000 ng·m L~(-1). The quantitation limit was 8 ng·m L~(-1).The intra-and inter-precision were ≤4. 20% and ≤5. 81%,the extraction recovery of ertapenem and internal standard were 96. 22%-100. 40%and 88. 29%-96. 50%,the intra-and inter-accuracy were-9. 30%-14. 00%,and the matrix effects of ertapenem and internal standard were 86. 10%-94. 87% and 90. 01%-97. 32%,respectively. All results of methodological study conformed to the requirements. Conclusion A reliable LC-MS/MS method with high sensitivity,simple sample preparation and short elution time was developed for the determination of ertapenem in mice plasma.
Objective To establish a rapid,sensitive method for the determination of ertapenem in mice plasma. Methods XZP-3408 was taken as the internal standard,the plasma samples were extracted by methanol to precipitate protein. After centrifugation,the supernatant was analyzed by LC-MS/MS. Chromatographic column of ZORBAX Eclipse plus C18( 4. 6 mm × 50. 0 mm,3. 5 micron),mobile phase composed of acetonitrile containing 0. 2% formic acid-water( 0. 2% formic acid)containing 10 mmol ·L~(-1) ammonium acetate in gradient elution mode,flow-rate of 0. 4 m L·min~(-1),injection volume of 5 μL. Electrospray positive ion detection method,multi-reaction monitoring. The specificity,standard curve and lower limit of quantification,precision and recovery,matrix effect,residual effect,dilution effect and stability of the method were investigated. Results Calibration curves were linear between 8 and 2000 ng·m L~(-1). The quantitation limit was 8 ng·m L~(-1).The intra-and inter-precision were ≤4. 20% and ≤5. 81%,the extraction recovery of ertapenem and internal standard were 96. 22%-100. 40%and 88. 29%-96. 50%,the intra-and inter-accuracy were-9. 30%-14. 00%,and the matrix effects of ertapenem and internal standard were 86. 10%-94. 87% and 90. 01%-97. 32%,respectively. All results of methodological study conformed to the requirements. Conclusion A reliable LC-MS/MS method with high sensitivity,simple sample preparation and short elution time was developed for the determination of ertapenem in mice plasma.
引文
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[11]刘秀菊,董维冲,王凌峰,等.大鼠血浆中厄他培南的HPLC法测定及初步药动学研究[J].药物分析杂志,2017,37(11):2082-2086.
[2]左震华,王东晓,管希周.厄他培南治疗中重度社区获得性肺炎疗效观察[J].国际呼吸杂志,2015,35(8):563-566.
[3]刘小平,王潇,刘泽滨,董晶.厄他培南对产超广谱β-内酰胺酶细菌的抑菌活性[J].中华医院感染学杂志,2009,19(20):2766-2768.
[4]周宇麒,张天托,谢灿茂,等.厄他培南静脉注射液治疗下呼吸道细菌性感染的有效性和安全性研究[J].实用医学杂志,2013,29(1):110-113.
[5]KOAL T,DETERS M,RESCH K,et al.Quantification of the carbapenem antibiotic ertapenem in human plasma by a validated liquid chromatography-mass spectrometry method[J].Clin Chim Acta,2006,364(1-2):239-245.
[6]LA MARCA G,GIOCALIERE E,VILLANELLI F,et al.Development of an UPLC-MS/MS method for the determination of antibiotic ertapenem on dried blood spots[J/OL].J Pharm Biomed Anal,2012,61:108-113.2012-03-05[2019-02-04].https://linkinghub.elsevier.com/retrieve/pii/S0731-7085(11)00725-4.
[7]MUNDKOWSKI R G,MAJCHER-PESZYNSKA J,BURKHARDTO,et al.A new simple HPLC assay for the quantification of ertapenem in human plasma,lung tissue,and broncho-alveolar lavage fluid[J].J Chromatogr B Analyt Technol Biomed Life Sci,2006,832(2):231-235.
[8]PICKERING M,BROWN S.Quantification and validation of HPLC-UV and LC-MS assays for therapeutic drug monitoring of erta-penem in human plasma[J].Biomed Chromatogr,2013,27(5):568-574.
[9]VAN RIJN S P,WESSELS A M A,GREIJDANUS B,et al.Quantification and validation of ertapenem using a liquid chromatographytandem mass spectrometry method[J].Antimicrob Agents Chemother,2014,58(6):3481-3484.
[10]CARDONE K E,GRABE D W,KULAWY R W,et al.Ertapenem pharmacokinetics and pharmacodynamics during continuous ambulatory peritoneal dialysis[J].Antimicrob Agents Chemother,2012,56(2):725-730.
[11]刘秀菊,董维冲,王凌峰,等.大鼠血浆中厄他培南的HPLC法测定及初步药动学研究[J].药物分析杂志,2017,37(11):2082-2086.