Quantitative evaluation of sphingomyelin and glucosylceramide using matrix-assisted laser desorption ionization time-of-flight mass spectrometry with sphingosylphosphorylcholine as an internal standard Practical application to tissues from patients with Niemann-Pick disease types A and C, and Gaucher disease
Quantitative evaluation of sphingomyelin and glucosylceramide using matrix-assisted laser desorption ionization time-of-flight mass spectrometry with sphingosylphosphorylcholine as an internal standard Practical application to tissues from patients with Niemann-Pick disease types A and C, and Gaucher disease
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DOI：
10.1016/j.jchromb.2008.05.013
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发表时间：
2008-07-15
影响因子：
3
通讯作者：
Yamaguchi, Seiji
中科院分区：
文献类型：
--
作者：
Fujiwaki, Takehisa;Tasaka, Masaru;Yamaguchi, Seiji
Niemann-Pick病A、C型和Gaucher病是糖脂储存障碍，其特征是糖鞘糖脂的系统性沉积，即Niemann-Pick病A、C型组织中的鞘磷脂和Gaucher病组织中的葡萄糖神经酰胺。用基质辅助激光解吸电离飞行时间质谱仪(MALDI-TOF/MS)分析了A、C型Niemann-Pick病和Gaucher病患者肝和脾组织中的鞘磷脂。用氯仿和甲醇从含有5 mg蛋白质的组织中提取粗脂。粗脂经温和碱处理后，制备了鞘磷脂部分，并进行了MALDI-TOF/MS分析。结果表明：(A)清晰地检测到不同种类的鞘磷脂和神经酰胺单己糖苷(CMH)的m/z值离子峰。(2)以鞘氨醇磷脂酰胆碱为定量内标物，计算了鞘磷脂和CMH的相对峰高，并绘制了它们与其含量的关系图。鞘磷脂和CMH的相对峰高分别在50~1500 ng和5~150 mg CMH含量范围内呈线性关系。(3)定量分析显示A、C型Niemann-Pick病患者肝、脾标本中神经鞘磷脂积聚，Gaucher病患者肝、脾标本中CMH显着积聚。本研究表明，用MALDI-TOF/MS方法可以定量检测鞘磷脂病患者少量组织中积聚的鞘磷脂和CMH。该方法不仅可用于各种鞘脂沉积症的诊断，而且可用于各种鞘脂沉积症患者的生化病理生理学评价。(C)2008 Elsevier B.V.，所有比赛已预订。
Niemann-Pick disease types A and C, and Gaucher disease are glycolipid storage disorders characterized by the systemic deposition of glycosphingolipids, i.e., sphingomyelin in Niemann-Pick disease types A and C tissues and glucosylceramide in Gaucher disease ones, respectively. Using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF/MS), we analyzed the sphingolipids in liver and spleen specimens from patients with Niemann-Pick disease types A and C, and Gaucher disease. Crude lipids were extracted from tissue containing 5 mg protein with chloroform and methanol. After mild alkaline treatment of the crude lipids, a sphingolipid fraction was prepared and analyzed by MALDI-TOF/MS. The results were as follows: (a) ion peaks with m/z values corresponding to different sphingomyelin and ceramide monohexoside (CMH) species were clearly detected. (b) With sphingosylphosphorylcholine as the internal standard for quantification of sphingomyelin and CMH, the relative peak heights of sphingomyelin and CMH were calculated and plotted versus their contents. The relative peak heights of sphingomyelin and CMH showed linearity between 50 and 1500 ng sphingomyelin content, and between 5 and 150 Fig CMH content, respectively. (c) Quantitative analysis revealed the accumulation of sphingomyelin in the liver and spleen specimens from the patients with Niemann-Pick disease types A and C. Striking accumulation of CMH was also detected in the liver and spleen specimens from the patients with Gaucher disease. This investigation indicated that accumulated sphingomyelin and CMH in small amounts of tissues from sphingolipidosis patients can be detected quantatively with the MALDI-TOF/MS method. This method will be useful not only for the diagnosis but also for biochemical pathophysiology evaluation of patients with various sphingolipidosis. (C) 2008 Elsevier B.V. All Fights reserved.