Dobbie M S, Surtees R A
Division of Biochemistry and Genetics, Institute of Child Health (UCLMS), London, UK.
J Chromatogr B Biomed Sci Appl. 1997 Aug 15;696(1):53-8. doi: 10.1016/s0378-4347(97)00221-1.
A simple method for the determination of the excitotoxin, quinolinic acid (QUIN) in cerebrospinal fluid (CSF) is described. QUIN, in lyophilized samples, was silylated by N-methyl-N-(tert.-butyldimethylsilyl)trifluoroacetamide in a single-step reaction at 65 to 70 degrees C to form a di-tert.-butyldimethylsilyl ester. Neither pre-purification of QUIN from CSF nor post-derivatisation sample clean-up was required. The derivatives were analysed by gas chromatography-electron impact mass spectrometry resulting in a prominent and characteristic [M-57]+ fragment ion which was used for quantitation. 2,6-Pyridine dicarboxylic acid, a structural analog of QUIN, was used as the internal standard. The detection limits for injected standards are in the femtomole range. CSF QUIN was found to be age-related and three preliminary reference ranges for CSF QUIN were found: 0 to 1 years, 31 +/- 15 nM QUIN (mean +/- standard deviation): 1.1 to 3 years, 26 +/- 15 nM; 3.1 to 14 years, 14 +/- 9 nM.
本文描述了一种测定脑脊液(CSF)中兴奋性毒素喹啉酸(QUIN)的简单方法。冻干样品中的QUIN在65至70摄氏度下,通过N-甲基-N-(叔丁基二甲基硅烷基)三氟乙酰胺一步反应进行硅烷化,形成二叔丁基二甲基硅烷基酯。既不需要从脑脊液中预先纯化QUIN,也不需要在衍生化后进行样品净化。通过气相色谱-电子轰击质谱法分析衍生物,得到一个突出且具有特征性的[M-57]+碎片离子,用于定量分析。2,6-吡啶二甲酸作为QUIN的结构类似物用作内标。注入标准品的检测限在飞摩尔范围内。发现脑脊液QUIN与年龄相关,并得出了脑脊液QUIN的三个初步参考范围:0至1岁,31±15 nM QUIN(平均值±标准差);1.1至3岁,26±15 nM;3.1至14岁,14±9 nM。