【摘要】 目的:基于RRLC-QQQ建立脑缺血后脑组织中未衍生化的内源性氨基酸的含量测定方法。方法:采用DiamonsilC18(4.6 mm×250 mm,5μm)色谱柱,15 min内同时测定12种氨基酸。乙腈-0.1%甲酸水梯度洗脱,流速为0.5 mL.min-1。离子源为ESI,正离子扫描模式,扫描方式为多反应检测。结果:各氨基酸标准曲线在检测范围内均呈良好线性(r>0.996),其检测限低于1.53μg.L-1,定量限低于3.09μg.L-1,高中低3个浓度水平日内和日间精密度的RSD均小于11%。结论:实际样品测定结果表明与假手术组相比,在脑缺血病理状态下,除N-乙酰天门冬氨酸外,脑组织中其余氨基酸水平均升高。部分氨基酸有显著变化,变化水平在术后12,24 h呈时间依赖性。该方法简便,快速,灵敏。这种方法可以应用于寻找脑缺血生物代谢标记物,探索研究脑缺血分子机制以及中药多成分组合药物协同作用机制。更多还原

【Abstract】 Objective: To establish a method to determine underivatized endogenous amino acids in brain tissues after cerebral ischemia based on RRLC-QQQ.Method: Diamonsil chromatographic column C18(4.6 mm×250 mm,5 μm) was adopted to determine 12 amino acids in 15 min,with acetonitrile-0.1% formic acid for gradient elution.The flow rate was set at 0.5 mL·min-1.With ESI as the ion source,positive ion scanning mode was adopted for multi-reaction monitoring.Result: Each amino acid standard curve(AAs) showed good linear relationship within the detection range(r>0.996),with the limit of detection of less than 11%,the limit of quantitation of less than 3.09 μg·L-1.The RSD of intra-and inter-day precisions at high,middle and low concentrations were less than 11%.Conclusion: The determination results of actual samples showed that compared with the levels of AAs of the sham operation group,all of the remaining amino acids apart from N-acetyl-aspartate increased in brain tissues.Some amino acids showed significant changes in a time-dependent manner after the operation.The method is so simple,rapid and sensitive that it can be used for finding biological metabolite markers of cerebral ischemia,and exploring cerebral ischemia molecular mechanisms and synergistic mechanism of combined administration of multi-component traditional Chinese medicines.更多还原