LESA 质谱代谢组学法筛查新生儿代谢缺陷症

Anal Chem. 2012 Nov 20;84(22):10113-20. doi: 10.1021/ac302527m. Epub 2012 Nov 2.

Metabonomics of newborn screening dried blood spot samples: a novel approach in the screening and diagnostics of inborn errors of metabolism.

新生儿干血斑样本的代谢组学研究：
一种新型的先天性代谢缺陷症的筛查及诊断方法

Dénes J, Szabó E, Robinette SL, Szatmári I, Szőnyi L, Kreuder JG, Rauterberg EW, Takáts Z.         
Biomolecular Medicine, Department of Surgery and Cancer, Imperial College London , United Kingdom.

英国伦敦帝国理工学院外科及癌症系生物分子医学部

Abstract 摘要

A novel, single stage high resolution mass spectrometry-based method is presented for the population level screening of inborn errors of metabolism. The approach proposed here extends traditional electrospray tandem mass spectrometry screening by introducing nanospray ionization and high resolution mass spectrometry, allowing the selective detection of more than 400 individual metabolic constituents of blood including acylcarnitines, amino acids, organic acids, fatty acids, carbohydrates, bile acids, and complex lipids. Dried blood spots were extracted using a methanolic solution of isotope labeled internal standards, and filtered extracts were electrosprayed using a fully automated chip-based nanospray ion source in both positive and negative ion mode. Ions were analyzed using an Orbitrap Fourier transformation mass spectrometer at nominal mass resolution of 100 000. Individual metabolic constituents were quantified using standard isotope dilution methods. Concentration threshold (cutoff) level-based analysis allows the identification of newborns with metabolic diseases, similarly to the traditional electrospray tandem mass spectrometry (ESI-MS/MS) method; however, the detection of additional known biomarkers (e.g., organic acids) results in improved sensitivity and selectivity. The broad range of detected analytes allowed the untargeted multivariate statistical analysis of spectra and identification of additional diseased states, therapeutic artifacts, and damaged samples, besides the metabolic disease panel.

本文介绍了一种新的单级高分辨质谱技术用于新生儿先天性代谢缺陷人群的筛查方法。该方法通过引入纳喷离子化及高分辨质谱技术来扩展传统电喷雾串联质谱在先天性代谢缺陷筛查方面的应用，它可以选择性检测超过400个不同的代谢产物，包括酰肉碱，氨基酸，有机酸，脂肪酸，碳水化合物，胆汁酸和复杂的脂类。采用含有同位素标记的内标的甲醇溶液对干血斑进行萃取，萃取液经过滤并通过全自动的芯片纳喷离子源纳喷，进行正、负离子模式分析。本文试验中采用了质量分辨率为十万（100,000）的傅里叶变换轨道阱质谱仪Orbitrap进行分析，并通过同位素稀释质谱法对不同的代谢成分进行定量分析。相似于传统电喷雾串联质谱法，基于浓度阈值（临界）水平的分析可以鉴定新生儿代谢疾病。然而，对于一些已知生物标记物（如有机酸），该方法表现出了更好的灵敏度和选择性。鉴于其对宽范围多组分的检测特征，该方法不仅可以用于代谢疾病筛查，还可以用于非特异性的谱图多元统计分析和对发生额外病变、人为治疗偏差、及受损样本的鉴定。