Iwamori M, Costello C, Moser H W
J Lipid Res. 1979 Jan;20(1):86-96.
Reaction of ceramides containing nonhydroxy fatty acids with benzoyl chloride in pyridine at 70 degrees C for 1 hr resulted in N-benzoylation to form N,N-acyl,benzoyl derivatives; O-benzoylation also occurred. However with ceramides containing 2-hydroxy fatty acids and phytosphingosine only O-benzoylation occurred even on prolonged treatment. Only O-benzoylation occurred on reaction with benzoic an hydride. However, the benzoylation of ceramides with phytosphingosine could not be achieved with benzoic anhydride and this benzoylation was performed by reaction with benzoyl chloride at 70 degrees C for 4 hr. Because N,N-acyl,benzoyl derivatives of ceramides containing nonhydroxy fatty acids produced by treatment with benzoyl chloride overlap methyl benzoate on high-performance liquid chromatography, benzoic anhydride was preferable for benzoylation of ceramides with nonhydroxy and 2-hydroxy fatty acids. On the other hand, the reaction with benzoyl chloride at 70 degrees C for 4 hr was used for quantitation of benzoylated ceramides containing 2-hydroxy fatty acids and phytosphingosine. 3-(p-Phenylbenzoyl)estrone was used as an internal standard for both reactions and values for ceramides containing 2-hydroxy fatty acids obtained by the two reactions were in good agreement. This procedure was applied to measurement of the ceramide levels in the brain, liver, and kidney of rats during development. The levels of ceramides containing nonhydroxy and 2-hydroxy fatty acids in the brain, liver, and kidney increased to the adult levels and then remained unchanged. Ceramide with phytosphingosine was detected in the liver and kidney, where its concentration gradually increased with age, but it was not found in the brain. The composition of nonhydroxy fatty acids were also analyzed.
含非羟基脂肪酸的神经酰胺与苯甲酰氯在吡啶中于70℃反应1小时，会发生N - 苯甲酰化反应，生成N,N - 酰基、苯甲酰基衍生物；同时也会发生O - 苯甲酰化反应。然而，含2 - 羟基脂肪酸的神经酰胺和植物鞘氨醇即使经过长时间处理，也仅发生O - 苯甲酰化反应。与苯甲酸酐反应时，仅发生O - 苯甲酰化反应。但是，植物鞘氨醇神经酰胺与苯甲酸酐无法实现苯甲酰化反应，而该苯甲酰化反应是通过与苯甲酰氯在70℃反应4小时来进行的。由于用苯甲酰氯处理产生的含非羟基脂肪酸的神经酰胺的N,N - 酰基、苯甲酰基衍生物在高效液相色谱上与苯甲酸甲酯重叠，所以苯甲酸酐更适合用于含非羟基和2 - 羟基脂肪酸的神经酰胺的苯甲酰化反应。另一方面，在70℃与苯甲酰氯反应4小时用于定量含2 - 羟基脂肪酸和植物鞘氨醇的苯甲酰化神经酰胺。3 - (对苯基苯甲酰基)雌酮用作这两种反应的内标，通过这两种反应获得的含2 - 羟基脂肪酸的神经酰胺的值吻合良好。该方法应用于发育过程中大鼠脑、肝和肾中神经酰胺水平的测定。脑、肝和肾中含非羟基和2 - 羟基脂肪酸的神经酰胺水平升高至成年水平后保持不变。在肝和肾中检测到了含植物鞘氨醇的神经酰胺，其浓度随年龄逐渐增加，但在脑中未发现。还分析了非羟基脂肪酸的组成。
