Application of 31 P NMR spectroscopy and chemical derivatization for metabolite profiling of lipophilic compounds in human serum

摘要

New methods for obtaining metabolic fingerprints of biological samples with improved resolution and sensitivity are highlysought 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 chemoselectiveprobes for improved resolution and quantitation is increasing. Considering the role of lipids in the pathogenesis of a numberof 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 with3113 and then detected with good resolution using 31 P NMR by exploiting the 100% natural abundance and wide chemical shiftrange of 31 P. After standardizing the reaction conditions using representative compounds, the derivatization approach wasused to profile lipids in human serum. The results show that the 31 P derivatization approach is simple, reproducible and highlyquantitative, and has the potential to profile a number of important lipids in complex biological samples.