测定脑脊液中兴奋性氨基酸的样品前处理方法

The Effect of Different Pretreatment on the Determination of Excitatory Amino Acids in Cerebrospinal Fluid Sample

采用高效液相色谱法，研究脑脊液样品不同的前处理方法对脑脊液中兴奋性氨基酸（谷氨酸、天门冬氨酶）测定结果的影响。结果表明：强酸性蛋白沉淀剂可导致脑脊液中的谷氨酸异常增高，脑脊液贮存于－８０℃并以甲醇作为蛋白沉淀剂时，兴奋氨基酸稳定，测定结果不变；如果脑脊液贮存在－２０℃，不能超过８周。建立了准确、灵敏的兴奋性氨基酸分析方法，为临床拮抗兴奋性毒性和进一步深入研究兴奋性氨基酸提供可靠的测定方法。

The changes of excitatory amino acids (EAAs, including glutamate and aspartate), in cerebrospinal fluid (CSF) were investigated by of different pretreatment methods. High performance liquid chromatography was employed for the determination of EA-As. It was found that deproteinization by precipitation with strong acid could lead to falsely high measurements of glutamate and when CSF was stored at -80degreesC and deproteinized with methanol before assay the EAAs in cerebrospinal fluid kept constant. It was suggested that the stored time of the sample could not be longer than 8 week when the cerebrospinal fluid was stored only at -20degreesC. An accurate and sensitive reversed-phase high performance liquid chromatographic method has been developed for anti-excitotoxity therapy and thorough study of EA-As in clinical.