FLUOROMETRIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF N-ACETYLNEURAMINIC AND N-GLYCOLYLNEURAMINIC ACIDS AND ITS APPLICATION TO THEIR MICRODETERMINATION IN HUMAN AND ANIMAL SERA, GLYCOPROTEINS, AND GLYCOLIPIDS
FLUOROMETRIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY OF N-ACETYLNEURAMINIC AND N-GLYCOLYLNEURAMINIC ACIDS AND ITS APPLICATION TO THEIR MICRODETERMINATION IN HUMAN AND ANIMAL SERA, GLYCOPROTEINS, AND GLYCOLIPIDS
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DOI：
10.1016/0003-2697(87)90377-0
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发表时间：
1987-07-01
影响因子：
2.9
通讯作者：
OHKURA, Y
中科院分区：
文献类型：
--
作者：
HARA, S;TAKEMORI, Y;OHKURA, Y
建立了一种简便、快速、高灵敏度的高效液相色谱法，用于测定人和动物血清、糖蛋白和糖脂中的N-乙酰和N-羟基神经氨酸。这些生物样品酸解释放的神经氨酸与α-酮酸的荧光试剂1，2-二氨基-4，5-亚甲二氧基苯在稀硫酸中转化为高荧光衍生物。衍生物在12分钟内在反相柱(Radial-Pak色谱柱C18)上用等浓度洗脱和荧光法检测。在10微升进样体积中，N-乙酰神经氨酸的检出限为25fmol(7.7pg)，N-羟基神经氨酸的检出限为23fmol(7.5pg)，信噪比为2。本方法可准确地测定人和动物血清中的神经氨酸，或0.25-2.5ug的糖蛋白和糖脂。
A simple, rapid, and highly sensitive high-performance liquid chromatographic method is described for the determination of N-acetyl- and N-glycolylneuraminic acids in human and animal sera, glycoproteins, and glycolipids. The neurmaninic acids, released by acid hydrolysis of these biological samples, are converted in dilute sulfuric acid with 1,2-diamino-4,5-methylenedioxybenzene, a fluorogenic reagent for .alpha.-keto acids, to highly fluorescent derivatives. The derivatives are separated within 12 min on a reversed-phase column (Radial-Pak cartridge C18) with an isocratic elution and detected fluorometrically. The detection limits are 25 fmol (7.7 pg) for N-acetylneuraminic acid and 23 fmol (7.5 pg) for N-glycoly neuraminic acid in a 10-.mu.l injection volume at a signal-to-noise ratio of 2. This method permits precise determination of the neuraminic acids in 5 .mu.l of human and animal sera or in 0.25-2.5 .mu.g of glycoprotein and glycolipids.