一种用于检测法伯病患者及对照者培养成纤维细胞中酸性神经酰胺酶活性的高效液相色谱分析方法。

A high-performance liquid chromatographic assay for acid ceramidase activity in cultured fibroblasts from patients with Farber's disease and from controls.

作者信息

Mitsuo K, Kobayashi T, Shinnoh N, Goto I

机构信息

Department of Neurology, Faculty of Medicine, Kyushu University, Fukuoka, Japan.

出版信息

Clin Chim Acta. 1988 Apr 29;173(3):281-7. doi: 10.1016/0009-8981(88)90015-0.

DOI:10.1016/0009-8981(88)90015-0

PMID:3383428

Abstract

The high-performance liquid chromatographic (HPLC) method we devised for assay of acid ceramidase activity involves coupling of a fluorescent probe to the enzymatically released sphingosine in the reaction mixture and detection of the fluorescent sphingosine derivative by reverse-phase HPLC. Using the method, acid ceramidase activity in fibroblast homogenates was accurately assayed, with or without the addition of exogenous ceramide, as the substrate, and the patients and carriers of Farber's disease could be readily diagnosed.

摘要

我们设计的用于测定酸性神经酰胺酶活性的高效液相色谱（HPLC）方法，包括将荧光探针与反应混合物中酶促释放的鞘氨醇偶联，并通过反相HPLC检测荧光鞘氨醇衍生物。使用该方法，可以准确测定成纤维细胞匀浆中的酸性神经酰胺酶活性，无论是否添加外源性神经酰胺作为底物，并且可以很容易地诊断出法伯病的患者和携带者。