Newborn Screening and Molecular Biology Branch, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
Clin Chim Acta. 2012 Aug 16;413(15-16):1217-21. doi: 10.1016/j.cca.2012.03.026. Epub 2012 Apr 4.
X-linked adrenoleukodystrophy (X-ALD) is the most common human peroxisomal disorder, and is caused by mutations in the peroxisomal transmembrane ALD protein (ALDP, ABCD1). The biochemical defect associated with X-ALD is an accumulation of very long-chain fatty acids (VLCFA, e.g. C24:0 and C26:0), which has been shown to result in the accumulation of C26:0-lysophosphatidylcholine (C26:0-LPC).
We describe the analysis of C26:0-LPC in dried-blood spots (DBS) using a rapid (30 min) and simple extraction procedure, isocratic HPLC resolution of LPC, and structure-specific analysis via negative ion mode tandem mass spectrometry.
In putative normal DBS specimens from newborns (N=223) C26:0-LPC was 0.09±0.03 μmol/l whole blood, while in peroxisomal biogenesis disorder (including X-ALD) patients (N=28) C26:0-LPC was 1.13±0.67 μmol/l whole blood. Both multiple reaction monitoring and a neutral loss scan (225.1 Da) analysis of DBS were used to analyze LPC.
Compared to a previous report of C26:0-LPC analysis in DBS, the method described here is simpler, faster, and more structure-specific for LPC with C26:0 acyl chains.
X 连锁肾上腺脑白质营养不良（X-ALD）是最常见的人类过氧化物酶体疾病，由过氧化物酶体跨膜 ALD 蛋白（ALDP，ABCD1）的突变引起。与 X-ALD 相关的生化缺陷是极长链脂肪酸（VLCFA，例如 C24:0 和 C26:0）的积累，这已被证明导致 C26:0-溶血磷脂酰胆碱（C26:0-LPC）的积累。
我们描述了使用快速（30 分钟）和简单的提取程序、等度 HPLC 分离 LPC 以及通过负离子模式串联质谱进行结构特异性分析，在干血斑（DBS）中分析 C26:0-LPC。
在来自新生儿的假定正常 DBS 标本（N=223）中，C26:0-LPC 为 0.09±0.03 μmol/l 全血，而在过氧化物酶体生物发生障碍（包括 X-ALD）患者（N=28）中，C26:0-LPC 为 1.13±0.67 μmol/l 全血。DBS 的多重反应监测和中性丢失扫描（225.1 Da）分析均用于分析 LPC。
与之前在 DBS 中分析 C26:0-LPC 的报告相比，这里描述的方法更简单、更快，并且对具有 C26:0 酰基链的 LPC 更具结构特异性。
