Journal
JOURNAL OF PROTEOME RESEARCH
Volume 15, Issue 9, Pages 3432-3440Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.jproteome.6b00234
Keywords
metabolic profiling; metabonomics; NMR spectroscopy; pipeline development; infant; urine
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Funding
- Imperial-National Institute for Health Research (NIHR) Clinical Phenome Centre
- NIHR Imperial Biomedical Research Centre based at Imperial College Healthcare National Health Service (NHS) Trust
- Imperial College London
- Imperial College Stratified Medicine Graduate Training Program in Systems Medicine and Spectroscopic Profiling (STRATiGRAD)
- UK Medical Research Council (MRC) [MC_PC_12025]
- Bruker Biospin
- Waters Corporation
- Metabometrix
- Imperial College
- MRC [MC_PC_12025] Funding Source: UKRI
- British Heart Foundation [PG/13/49/30307] Funding Source: researchfish
- Medical Research Council [MC_PC_12025] Funding Source: researchfish
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Numerous metabolic profiling pipelines have been developed to characterize the composition of human biofluids and tissues, the vast majority of these being for studies in adults. To accommodate limited sample volume and to take into account the compositional differences between adult and infant biofluids, we developed and optimized sample handling and analytical procedures for studying urine from newborns. A robust pipeline for metabolic profiling using NMR spectroscopy was established, encompassing sample collection, preparation, spectroscopic measurement, and computational analysis. Longitudinal samples were collected from five infants from birth until 14 months of age. Methods of extraction and effects of freezing and sample dilution were assessed, and urinary contaminants from breakdown of polymers in a range of diapers and cotton wool balls were identified and compared, including propylene glycol, acrylic acid, and tert-butanol. Finally, assessment of urinary profiles obtained over the first few weeks of life revealed a dramatic change in composition, with concentrations of phenols, amino acids, and betaine altering systematically over the first few months of life. Therefore, neonatal samples require more stringent standardization of experimental design, sample handling, and analysis compared to that of adult samples to accommodate the variability and limited sample volume.
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