Poulos A, Pollard A C
Clin Chim Acta. 1976 Nov 1;72(3):327-35. doi: 10.1016/0009-8981(76)90195-9.
A method for the assay of leukocyte beta-galactocerebrosidase, beta-glucocerebrosidase and sphingomyelinase activities has been developed, based on the separation of the tritiated sphingolipid substrates from their corresponding radioactive hydrophobic product (ceramide) by thin-layer chromatography on Silica gel H coated microscope slides. For the determination of beta-galactocerebrosidase and beta-glucocerebrosidase activities the silica gel is impregnanted with sodium tetraborate. Each chromatogram is easily divided into two distinct zones and the radioactivity content of each is determined by liquid scintillation counting. The technique described, is rapid, less costly than conventional methods and provides an accurate assessment of sphingolipid hydrolase activity. It is suggested that it should be of considerable value in those areas which require the rapid analysis of large numbers of samples, such as in screening for the sphingolipidoses or for enzyme purification studies.
基于在硅胶H涂层显微镜载玻片上进行薄层色谱，将氚标记的鞘脂底物与其相应的放射性疏水产物（神经酰胺）分离，已开发出一种用于测定白细胞β-半乳糖脑苷脂酶、β-葡萄糖脑苷脂酶和鞘磷脂酶活性的方法。为了测定β-半乳糖脑苷脂酶和β-葡萄糖脑苷脂酶的活性，硅胶用四硼酸钠浸渍。每个色谱图很容易分成两个不同的区域，每个区域的放射性含量通过液体闪烁计数来确定。所描述的技术快速，比传统方法成本低，并能准确评估鞘脂水解酶活性。建议在那些需要快速分析大量样品的领域，如鞘脂沉积症筛查或酶纯化研究中，该技术具有相当大的价值。
