Current state of the art of analyte scope in urine metabolome analysis by non-hydrogenative PHIP

Abstract

Non-hydrogenative PHIP (nh-PHIP) is an NMR signal enhancement technique that offers several orders of magnitude gains in detection sensitivity. It is one of the few hyperpolarization methods that have been demonstrated to be applicable to chemical analysis of biological samples and potentially metabolomics. It is, however, a chemoselective method and needs to be tuned to particular analyte and metabolite classes at a time.Herein, we present a systematic study where we apply four nh-PHIP modifications to urine samples from two different species – human and dog. Firstly, this allows to explore the whole analyte class scope and present what information is nh-PHIP capable of providing by varying the composition of the nh-PHIP catalyst system and the sample preparation protocol. Secondly, comparing hyperpolarized spectra from urines from different species demonstrates that this hyperpolarization technique is robust and tolerant of possibly considerable matrix differences: signals of the same metabolites appear at same chemical shifts from urines that differ from one-another much more than is likely in a realistic metabolomics study. Thereby we propose the idea that nh-PHIP is ready for application in metabolomics experiments.

Keywords

• Biofluids
• Urine
• Metabolomics
• NMR
• Parahydrogen
• Hyperpolarization