LC-MS同时测定大鼠脑脊液中谷氨酸和γ-氨基丁酸的方法研究  

Simultaneous determination of glutamic acid and γ-aminobutyric acid in rat cerebrospinal fluid by LC-MS

作　　者：廖昌军[1] 臧志和[1] 王德艳[1] 

机构地区：[1]成都医学院药学院,四川成都610083

出　　处：《中国测试》2015年第2期50-53,共4页China Measurement & Test

基　　金：成都医学院药学重点学科建设项目(CYXK2012006)

摘　　要：建立LC-MS同时检测大鼠脑脊液中谷氨酸和γ-氨基丁酸的方法。采用安捷伦ZORBAX SIL（4.6 mm×250 mm×5μm）硅胶正相色谱柱,流动相为甲醇-水（0.1%甲酸）=50∶50（v/v）,梯度洗脱,流速为1 m L/min。电喷雾离子源（eletrospray ionization,ESI）采用正离子化方式,以多重反应检测（multiple reaction monitoring,MRM）方式进行检测,用于定量分析的离子反应分别为m/z 148.1→m/z 84.1（谷氨酸,[M＋H]＋）,m/z 104.1→m/z 87.1（γ-氨基丁酸,[M＋H]＋）。测定大鼠脑脊液中谷氨酸在500~2 500μg/L和γ-氨基丁酸在50~250μg/L质量浓度范围内,浓度与峰面积呈良好的线性关系。该方法简便、准确、灵敏度高,专属性强,重复性好,适用于脑脊液中谷氨酸和γ-氨基丁酸的同时测定,可为氨基酸类神经递质提供有效的检测手段。To establish LC-MS method to simultaneously determine the contents of glutamic acid and γ-aminobutyric acid in cerebrospinal fluid of rat. The Agilent ZORBAX SIL（4.6mm×250 mm× 5 μm） silica gel was chromatographic column maintained at 25℃. The mobile phase consisted of methanol water （0.1% formic acid）=50：50（v/v）, gradient elution. The flow rate was 1 mL/min. The glutamic acid and γ-aminobutyric acids were detected by LC-MS using positive ESI with MRM. The specificions （m/z） used for quantification were m/z 148.1→μm/z 84.1（Glu,[M＋H]＋）, m/z 104.1→m/z 87.1 （GABA,[M＋H]＋）. The calibration curve for Glutamic acid and γ-aminobutyric acid was linear in the range of 500 μg/L to 2 500 p,g/L, 50 μg/L to 250 μg/L, respectively. This method shows excellent separation, simply, accurately, high sensitivity, specificity, good repeatability. It is suitable for the separation and quantization of glutamic acid and ~/-aminobutyric acid in cerebrospinal fluid tissue, for which it also provide an available detection method.

关 键 词：液质联用法 谷氨酸 Γ-氨基丁酸 脑脊液 

分 类 号：O657.32[理学—分析化学]