CAJ | 학술논문

目的：建立 LC-MS／MS 法同时测定小鼠肝脏中10种游离型与结合型胆汁酸的浓度。方法肝组织样品匀浆经乙腈沉淀蛋白后进行 LC-MS／MS 分析，空白基质经活性炭（100 mg／mL）吸附获得，色谱柱为 Zorbax Eclipse XDB-C18柱（5μm，150 mm×4.60 mm），流动相为乙腈-含0.05％甲酸和4 mmol／L 乙酸铵溶液，梯度洗脱，流速1 mL／min，进样10μL，采用电喷雾离子源（ESI）、负离子的多级反应模式（MRM）监测。结果在定量范围内，各胆汁酸的线性关系良好，检测时间较短（14 min），精密度、准确度、提取回收率等均符合要求。结论建立的方法灵敏度高、专属性好，可用于肝组织样品中胆汁酸浓度的测定。
목적：건립 LC-MS／MS 법동시측정소서간장중10충유리형여결합형담즙산적농도。방법간조직양품균장경을정침정단백후진행 LC-MS／MS 분석，공백기질경활성탄（100 mg／mL）흡부획득，색보주위 Zorbax Eclipse XDB-C18주（5μm，150 mm×4.60 mm），류동상위을정-함0.05％갑산화4 mmol／L 을산안용액，제도세탈，류속1 mL／min，진양10μL，채용전분무리자원（ESI）、부리자적다급반응모식（MRM）감측。결과재정량범위내，각담즙산적선성관계량호，검측시간교단（14 min），정밀도、준학도、제취회수솔등균부합요구。결론건립적방법령민도고、전속성호，가용우간조직양품중담즙산농도적측정。
Objective To develop a high-performance liquid chromatography tandem mass spectrometry (LC-MS/MS)method for simultaneous determination of 10 kinds of free and conjugated bile acids in mice liver.Methods Liver samples (homogenized in deionized water)were precipitated as protein by acetonitrile (100mg/ml)to be analyzed by LC-MS/MS.Blank matrix was prepared with 100mg/ml activated charcoal.Analytes were separated on a Zorbax Eclipse XDB-C18 column (5μm,150mm ×4.60mm)with gradient elution of acetonitrile and 4mmol/L ammonium acetate in aqueous solution (contained 0.05% formic acid),of which flow rate was 1ml/min and injection volume was 10μl.Detection was carried out by multiple reaction monitoring (MRM)in negative ion mode.Results The method had a good linear relationship in quantitative range,with a short run time as 14min,of which precision,accuracy,extraction recovery and other parameters met requirement.Conclusion The proposed method,proved to be sensitive and selective,is suitable for concentration determination of bile acids in mouse liver.