Nishimura K, Nakamura A
J Biochem. 1985 Nov;98(5):1247-54. doi: 10.1093/oxfordjournals.jbchem.a135391.
A new sensitive assay method for sphingoids, 4-D-hydroxysphinganine, sphingosine, and sphinganine, involving reversed phase HPLC was described. Chromatography of p-N-nitrophenylacetyl derivatives of the sphingoids showed sufficient resolution, and each peak showed a linear relationship between molar quantity and area response, from 10 pmol to 10 nmol. The method was applied to the analysis of long chain bases in intestinal glycolipids from embryonic and adult Japanese quails. At the embryonic stage of 12 days' incubation, 4-D-hydroxysphinganine accounted for about 40% of the long chain bases of glycolipids, a concentration comparable to that in adult tissue. Egg yolk, which is an exclusive exogenous nutrient mixture supplied by the mother, contained only a few percent of 4-D-hydroxysphinganine as a glycolipid component. While, [3H]palmitic acid administered to embryos was incorporated into 4-D-hydroxysphinganine as well as sphingosine and sphinganine. These results suggest that sphingoids of intestinal glycolipids including 4-D-hydroxysphinganine are synthesized in the tissue, excluding the possibility of their exogenous origin in the embryonic tissue.
描述了一种用于鞘脂类、4-D-羟基鞘氨醇、鞘氨醇和二氢鞘氨醇的新型灵敏测定方法，该方法涉及反相高效液相色谱法。鞘脂类的对硝基苯乙酰衍生物的色谱分析显示出足够的分辨率，并且每个峰在10皮摩尔至10纳摩尔的摩尔量与面积响应之间呈现线性关系。该方法应用于分析来自胚胎期和成年日本鹌鹑肠道糖脂中的长链碱。在孵化12天的胚胎阶段，4-D-羟基鞘氨醇约占糖脂长链碱的40%，其浓度与成年组织中的浓度相当。蛋黄是由母体提供的唯一外源营养混合物，仅含有百分之几的4-D-羟基鞘氨醇作为糖脂成分。而给胚胎注射的[3H]棕榈酸会掺入到4-D-羟基鞘氨醇以及鞘氨醇和二氢鞘氨醇中。这些结果表明，包括4-D-羟基鞘氨醇在内的肠道糖脂鞘脂类是在组织中合成的，排除了它们在胚胎组织中外源来源的可能性。
