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高效液相色谱法在确定无菌性脑膜炎患者脑脊液中游离氨基酸模式异常方面的应用。

Use of high performance liquid chromatography in defining the abnormalities in the free amino acid patterns in the cerebrospinal fluid of patients with aseptic meningitis.

作者信息

Halawa I, Baig S, Qureshi G A

机构信息

Department of Neurology, Karolinska Institute, Huddinge University Hospital, Stockholm, Sweden.

出版信息

Biomed Chromatogr. 1991 Sep;5(5):216-20. doi: 10.1002/bmc.1130050508.

Abstract

Free amino acids were quantitatively determined in cerebrospinal fluid (CSF) and plasma samples from patients with aseptic meningitis by a newly developed high performance liquid chromatographic (HPLC) method. The method of analysis was based on precolumn derivatization of orthophthaladehyde in the presence of 2-mercaptoethanol and detection was made at Eex = 340 nm and Eem = 450 nm. The method was sensitive and the limit for detection was less than 1 pmol for most of the amino acids. It took 45 min to separate 26 amino acids with highly reproducible results, giving a coefficient of variance for retention times and integrated areas less than 0.4% and 2%, respectively, after five replicate runs. The results accumulated in 10 patients were compared statistically with 11 age-matched healthy controls. Among the amino acids almost all the neurotransmitter candidates, such as aspartic acid, glutamic acid, glutamine, glycine, tyrosine, phenylalanine and gamma-aminobutyric acid (GABA), were significantly increased in the patients' CSF, whereas arginine and threonine were low. No change was observed in plasma amino acids in patients as compared to healthy controls. The higher levels of most of the neurotransmitters, especially GABA, aspartic acid and glutamic acid, could be used diagnostically in assessing the progression and remission in aseptic meningitis.

摘要

采用新开发的高效液相色谱(HPLC)法对无菌性脑膜炎患者的脑脊液(CSF)和血浆样本中的游离氨基酸进行了定量测定。分析方法基于在2-巯基乙醇存在下邻苯二甲醛的柱前衍生化,并在激发波长(Eex)=340nm和发射波长(Eem)=450nm处进行检测。该方法灵敏度高,大多数氨基酸的检测限小于1皮摩尔。分离26种氨基酸耗时45分钟,结果具有高度重现性,五次重复运行后保留时间和积分面积的变异系数分别小于0.4%和2%。对10例患者积累的结果与11例年龄匹配的健康对照进行了统计学比较。在氨基酸中,几乎所有神经递质候选物,如天冬氨酸、谷氨酸、谷氨酰胺、甘氨酸、酪氨酸、苯丙氨酸和γ-氨基丁酸(GABA),在患者脑脊液中均显著升高,而精氨酸和苏氨酸含量较低。与健康对照相比,患者血浆氨基酸未见变化。大多数神经递质,尤其是GABA、天冬氨酸和谷氨酸的较高水平可用于诊断评估无菌性脑膜炎的进展和缓解情况。

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