Yu Zhanru, Huang Honglei, Zhang Hanlin, Kessler Benedikt M
TDI Mass Spectrometry Laboratory, Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7FZ, UK.
Kennedy Institute of Rheumatology, Nuffield Department of Orthopaedics, Rheumatology and Musculoskeletal Sciences, University of Oxford, Oxford OX3 7FY.
Talanta. 2019 Jul 1;199:184-188. doi: 10.1016/j.talanta.2019.02.062. Epub 2019 Feb 18.
Polyamines are a class of poly-cationic aliphatic amines, playing a role in different cellular processes such as maintaining intracellular pH and membrane potential that are relevant for general cellular physiology and ageing. The development of analytical methods for detection and quantitation of this class of compounds has been challenging due to the basic nature of these species. Both liquid chromatography (LC) and gas chromatography (GC) have been applied for separation, mostly coupled to mass spectrometry (MS) for detection. However, current methodologies suffer from lengthy extraction protocols and limitations in separation and detection levels. Here, we present a simplified and optimised method for straightforward extraction of polyamine metabolites including spermine, spermidine, norspermidine, cadaverine and putrescine from cellular and tissue material. We demonstrate that strong acid-based extraction and chemical derivatisation not only improves isolation, but also recovery. Combined with two-dimensional gas chromatography, this method provides clear separation and femtomole sensitivity for the profiling of polyamines.
多胺是一类多阳离子脂肪族胺，在维持细胞内pH值和膜电位等不同细胞过程中发挥作用，这些过程与一般细胞生理学和衰老相关。由于这类化合物的碱性性质，用于检测和定量此类化合物的分析方法的开发一直具有挑战性。液相色谱（LC）和气相色谱（GC）都已用于分离，大多与质谱（MS）联用进行检测。然而，目前的方法存在提取方案冗长以及分离和检测水平有限的问题。在此，我们提出一种简化且优化的方法，用于从细胞和组织材料中直接提取多胺代谢物，包括精胺、亚精胺、 norspermidine、尸胺和腐胺。我们证明基于强酸的提取和化学衍生化不仅能改善分离，还能提高回收率。结合二维气相色谱，该方法为多胺分析提供了清晰的分离和飞摩尔灵敏度。
