Nagai Yuko, Sakakibara Iori, Toyoda Hidenao
Faculty of Pharmaceutical Sciences, Ritsumeikan University.
Anal Sci. 2019 May 10;35(5):517-520. doi: 10.2116/analsci.18P500. Epub 2018 Dec 28.
An analytical method for the determination of sialic acids in biological samples has been developed and applied to fetal bovine serum (FBS), newborn calf serum and adult bovine serum. The hydrolysis of sera was carried out and the liberated sialic acids were quantified using a rapid and sensitive HPLC. The HPLC includes the separation and detection of N-acetylneuraminic acid (Neu5Ac) and N-glycolylneuraminic acid (Neu5Gc) using hydrophilic interaction liquid chromatography and a fluorometric post-column reaction with 2-cyanoacetamide. The calibration graphs for Neu5Ac and Neu5Gc were linear over the range of 10 pmol - 5 nmol. The concentrations of sialic acids in FBS, newborn calf serum and adult bovine serum were 5.06, 3.79 and 1.64 mM, respectively. The ratios of Neu5Gc and Neu5Ac changed dramatically according to the development stages. The present method has a satisfactory sensitivity in the quantification of Neu5Ac and Neu5Gc in serum samples. It seems that this analytical system can therefore be applied for routine use in clinical investigations of serum sialylation changes in cancer patients.
已开发出一种用于测定生物样品中唾液酸的分析方法，并将其应用于胎牛血清（FBS）、新生牛血清和成年牛血清。对血清进行水解，使用快速灵敏的高效液相色谱法对释放出的唾液酸进行定量。该高效液相色谱法包括使用亲水相互作用液相色谱法分离和检测N-乙酰神经氨酸（Neu5Ac）和N-羟乙酰神经氨酸（Neu5Gc），以及与2-氰基乙酰胺进行柱后荧光反应。Neu5Ac和Neu5Gc的校准曲线在10 pmol - 5 nmol范围内呈线性。FBS、新生牛血清和成年牛血清中唾液酸的浓度分别为5.06、3.79和1.64 mM。Neu5Gc与Neu5Ac的比例根据发育阶段有显著变化。本方法在血清样品中Neu5Ac和Neu5Gc的定量方面具有令人满意的灵敏度。因此，这种分析系统似乎可用于癌症患者血清唾液酸化变化的临床研究中的常规检测。
