NPC publishes UPLC-MS methods for profiling of urine for large-scale metabolic profiling

The NPC has published a paper detailing its analytical and preparative procedures for metabolic phenotyping of human urine by UPLC-MS.

Following the 2014 publication of standardised methods for NMR analysis of biofluids on a large- scale, the Mass Spectrometry team have also now developed the first suite of UPLC-MS assays suitable for large-scale metabolic profiling and this publication marks the significant step of making this widely available for the metabolic profiling community. To better understand the molecular mechanisms underpinning physiological variation in human populations, metabolic phenotyping approaches are increasingly being applied to studies involving hundreds and thousands of biofluid samples. Hyphenated ultra-performance liquid chromatography- mass spectrometry (UPLC-MS) has become a fundamental tool for this purpose. However, the seemingly inevitable need to analyse large studies in multiple analytical batches for UPLC-MS

analysis poses a challenge to data quality which has been recognized in the field. The National Phenome Centre has developed a fit-for-purpose UPLC-MS platform, method set, and sample analysis workflow, capable of sustained analysis on an industrial scale and allowing batch-free operation for large studies. Using complementary reversed-phase chromatography (RPC) and hydrophilic interaction liquid chromatography (HILIC) together with high resolution orthogonal acceleration time-of-flight mass spectrometry (oaTOF-MS), exceptional measurement precision is exemplified with independent epidemiological sample sets of approximately 650 and 1000 participant samples. The results in this paper demonstrate that the platform and methodology here at the National Phenome Centre is fit-for-use in large scale metabolic phenotyping studies, challenging the assertion that such screening is inherently limited by batch effects.

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