Yu R K, Ledeen R W
J Lipid Res. 1970 Nov;11(6):506-16.
A method is described for analysis of gangliosides by GLC assay of the sialic acid component. Mild acid treatment in methanol converted the latter to methyl ketoside methyl ester, which was then chromatographed as the TMS derivative. The major methanolysis product was shown to be the beta-anomer, and its chromatographic peak was used for quantification. NANA and NGNA could be analyzed simultaneously, while an O-acetylated derivative of NGNA was detected qualitatively. Standard curves were obtained for the three following representative samples: (a) a mixture of beef brain gangliosides, (b) Tay-Sachs ganglioside, and (c) hematoside-NANA. These had different slopes which reflected the variation in yield of beta-NANA obtained from methanolysis. The smallest sample analyzed in the present study contained 0.3 micro g of NANA. The advantage of GLC in solving the problem of false chromogens is illustrated in a comparative study with two colorimetric procedures. Two columns are described whose combined use is highly effective in establishing identity and in eliminating false peaks when they arise. The GLC method has been applied to analysis of the total brain ganglioside content of several species, and a general trend was observed toward decreasing levels in the lower vertebrates. In addition, NGNA was detected and quantified in several of these samples.
本文描述了一种通过对唾液酸成分进行气相色谱分析来测定神经节苷脂的方法。在甲醇中进行温和的酸处理可将后者转化为甲基酮糖苷甲酯，然后将其作为三甲基硅烷基（TMS）衍生物进行色谱分析。结果表明，主要的甲醇解产物是β-异头物，其色谱峰用于定量分析。N-乙酰神经氨酸（NANA）和N-羟乙酰神经氨酸（NGNA）可同时进行分析，同时对NGNA的O-乙酰化衍生物进行了定性检测。针对以下三种代表性样品获得了标准曲线：（a）牛脑神经节苷脂混合物，（b）泰-萨克斯神经节苷脂，以及（c）血苷-NANA。这些标准曲线具有不同的斜率，反映了甲醇解得到的β-NANA产量的变化。本研究中分析的最小样品含有0.3微克的NANA。在与两种比色法的对比研究中，展示了气相色谱法在解决假色原问题方面的优势。描述了两根色谱柱，当出现假峰时，联合使用这两根色谱柱在确定物质同一性和消除假峰方面非常有效。气相色谱法已应用于几种物种全脑神经节苷脂含量的分析，并观察到一个总体趋势，即低等脊椎动物中的含量逐渐降低。此外，在其中几个样品中检测并定量了NGNA。
