Department of Chemistry, Purdue University, 560 Oval Drive, West Lafayette, IN 47907, USA.
Magn Reson Chem. 2009 Dec;47 Suppl 1(Suppl 1):S74-80. doi: 10.1002/mrc.2480.
New methods for obtaining metabolic fingerprints of biological samples with improved resolution and sensitivity are highly sought for early disease detection, studies of human health and pathophysiology, and for better understanding systems biology. Considering the complexity of biological samples, interest in biochemical class selection through the use of chemoselective probes for improved resolution and quantitation is increasing. Considering the role of lipids in the pathogenesis of a number of diseases, in this study fingerprinting of lipid metabolites was achieved by (31)P labeling using the derivatizing agent 2-chloro-4,4,5,5-tetramethyldioxaphospholane. Lipids containing hydroxyl, aldehyde and carboxyl groups were selectively tagged with (31)P and then detected with good resolution using (31)P NMR by exploiting the 100% natural abundance and wide chemical shift range of (31)P. After standardizing the reaction conditions using representative compounds, the derivatization approach was used to profile lipids in human serum. The results show that the (31)P derivatization approach is simple, reproducible and highly quantitative, and has the potential to profile a number of important lipids in complex biological samples.
新方法用于获得具有改进分辨率和灵敏度的生物样品代谢指纹，这对于早期疾病检测、人类健康和病理生理学研究以及更好地理解系统生物学非常重要。考虑到生物样品的复杂性，人们越来越感兴趣通过使用化学选择性探针来改善分辨率和定量来进行生化类别选择。鉴于脂质在许多疾病发病机制中的作用，在这项研究中，通过使用衍生试剂 2-氯-4,4,5,5-四甲基二氧杂环戊磷烷进行（31）P 标记来实现脂质代谢物指纹图谱。含有羟基、醛基和羧基的脂质被选择性地标记上（31）P，然后通过利用（31）P 的 100%天然丰度和宽化学位移范围，使用（31）P NMR 以良好的分辨率进行检测。在用代表性化合物标准化反应条件后，该衍生化方法用于分析人血清中的脂质。结果表明，（31）P 衍生化方法简单、可重现且高度定量，并且有可能对复杂生物样品中的许多重要脂质进行分析。
