摘要
目的建立液相色谱串联质谱法检测大鼠海马和纹状体中氨基酸类神经递质含量的方法。方法选用Kromasil C18(250 mm×4.6mm,5μm)色谱柱,以0.1%甲酸-乙腈(90∶10)为流动相,流速0.6 mL/min。离子源为电喷雾电离源(electrospray ionization,ESI),正离子化方式,以多重反应监测(multiple reaction monitoring,MRM)模式进行扫描定量,用于定量分析的离子对分别m/z148→m/z84(Glu),m/z134→m/z74(Asp),m/z104→m/z87(GABA),m/z76→m/z30(Gly)。测定SD大鼠海马和纹状体中4种氨基酸类神经递质的浓度。结果 4种氨基酸在4.8 min内即可完成测定,Glu在103～12 875 ng/mL线性关系良好,Asp和GABA在102～12 750 ng/mL线性关系良好,Gly在61.20～7 650 ng/mL线性关系良好。各神经递质的加样回收率均在91%～107%(RSD≤9.12%)。结论本方法简便、准确、灵敏度高,专属性强,重复性好,适用于脑组织中氨基酸类神经递质的测定,可为氨基酸类神经递质提供有效的检测手段。
Objective To establish LC-ESI-MS/MS method to simultaneously determine the contents of glutamic acid,aspartic acid,γ-aminobutyric acid,and glycine in hippocampus and corpus striatum of rat.Methods Kromasil C18 column（250 mm×4.6 mm,5 μm） maintained at 25 ℃ with 0.1% formic acid-acetonitrile（90∶10） as the mobile phase was used.The flow rate was 0.6 mL/min.The amino acids were detected by MS/MS using positive ESI with MRM.The specificions（m/z） used for quantification were m/z148→m/z84（Glu）,m/z134→m/z74（Asp）,m/z104→m/z87（GABA）,m/z76→m/z30（Gly）.Results The neurotransmitter amino acids were separated in 4.8 min.The calibration curve for glutamic acid,aspartic acid,γ-aminobutyric acid and glycine was linear in the range of 103~12 875 ng/mL,102~12 750 ng/mL,102~12 750 ng/mL and 61.20~7 650 ng/mL,respectively.The average recovery was among 91%~107%（RSD≤9.12%）.Conclusion This method shows excellent separation,linearity,high sensitivity,specificity,good repeatability.It is suitable for the separation and quantization of amino acids in brain tissue,for which it also provide an available detection method.
出处
《今日药学》
CAS
2013年第2期65-69,共5页
Pharmacy Today
基金
国家自然科学基金资助项目(编号:30772756)
