Nakabayashi H, Iwamori M, Nagai Y
J Biochem. 1984 Oct;96(4):977-84.
A highly sensitive method for the analysis and quantitation of gangliosides by high-performance liquid chromatography is described. The method involves simple, quantitative and specific derivatization of gangliosides, involving their carboxylic acid groups, to UV-absorbing p-bromophenacyl derivatives. The molar extinction coefficient of the derivatives at maximum absorption at 261 nm was about 23,000. The acidic fraction separated by DEAE-Sephadex column chromatography was directly derivatized and the reaction mixture was directly injected into a high-performance liquid chromatograph without any purification of the derivatives. Monosialogangliosides, GM4, GM3, GM2, and GM1, were well separated by normal phase high-performance liquid chromatography. The standard curve for quantitation of gangliosides was linear up to 100 micrograms of ganglioside-sialic acid and the lower limit of detection was about 10 ng. Also, by reverse phase high performance liquid chromatography, the molecular species of gangliosides were successfully analyzed. The method was shown to be applicable to the analysis of gangliosides in biological materials.
描述了一种通过高效液相色谱法分析和定量神经节苷脂的高灵敏度方法。该方法涉及神经节苷脂的简单、定量和特异性衍生化，即利用其羧酸基团将其转化为具有紫外吸收的对溴苯甲酰衍生物。衍生物在261nm最大吸收波长处的摩尔消光系数约为23000。通过DEAE-葡聚糖柱色谱分离得到的酸性组分直接进行衍生化，反应混合物无需对衍生物进行任何纯化即可直接注入高效液相色谱仪。单唾液酸神经节苷脂GM4、GM3、GM2和GM1通过正相高效液相色谱法得到了很好的分离。神经节苷脂定量的标准曲线在高达100μg神经节苷脂-唾液酸的范围内呈线性，检测下限约为10ng。此外，通过反相高效液相色谱法成功分析了神经节苷脂的分子种类。该方法被证明适用于生物材料中神经节苷脂的分析。
