摘要
目的建立液相色谱-串联质谱法(LC-MS/MS)测定人尿中乳果糖、甘露醇和乳糖含量。方法尿样用水和乙腈直接稀释后,以LunaNH2(100×2.0mm,5μm)为分析柱,乙腈-水(80∶20,v/v)为流动相,0.3mL/min流速进行色谱分离。采用负离子化电喷雾、多反应监测(MRM)扫描方式进行质谱定量分析,离子通道选择m/z为:甘露醇181.1/88.9,乳果糖和乳糖341.3/160.9,内标353.4/166.9。结果甘露醇、乳果糖、乳糖和内标的保留时间分别为2.9、4.1、4.7和4.7min。在0.1～40μg/mL浓度范围内线性关系良好(r≥0.9980)。最低定量限分别为0.1μg/mL(甘露醇、乳果糖)和0.2μg/mL(乳糖)。日内、日间精密度(CV%)分别为0.60%～4.31%、2.55%～7.71%,准确度分别为86.4%～107.6%、95.9%～102.9%。回收率为84.7%～105.5%。结论该方法的样品处理简单快速,检测专一灵敏、准确可靠,可用于肠通透性评价中甘露醇及乳果糖的含量测定。
[Objective] To establish a LC-MS/MS method for the determination of lactulose, mannitol and lactose in human urine. [Method] Urine samples were simply diluted with water and acetonitrile. Chromatographic separa- tion was performed on Phenomenex Luna NH2 （100×2.0 mm, 5μm） column using acetonitrile-water （80：20, v/v） as mobile phase at flow rate of 0.3 mL/min. The analysis were protonated in the positive electrospray ionization （ESI） interface and detected in multiple reaction monitoring （MRM） mode with the transitions from the precursor to the production of m/z 181.1/88.9 for mannitol, m/z 341.3/160.9 for lactulose and lactose and ndz 353.4/166.9 for internal standard （IS）. [Results] The retention time of mannitol, lactulose, lactose and IS were found at 2.9, 4.1, 4.7 and 4.7 rain. Calibration curve was linear in the range from 0.1 to 40 μg/mL with a correlation coefficient above 0.9980. The lower limit of quantitation was 0.1 μg/mL for mannitol and lactulose, 0.2 μg/mL for lactose. The intra- and interday precision （CV%） were 0.60%-4.31% and 2.55%-7.71%, the intra- and inter-day accuracy were 86.4%-107.6% and 95.9%-102.9%, the recovery was 84.7%-105.5%. [Conclusion] The LC-MS/MS method for the quantitative determination of mannitol, lactulose and lactose in human urine was simple, fast, sensitive and specific, which can be used for mannitol and lactulose determination for intestinal permeability assessment in clinic applications.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第13期1810-1813,1817,共5页
China Journal of Modern Medicine
