A GC-TOF-MS study of the stability of serum and urine metabolomes during the UK Biobank sample collection and preparation protocols

Abstract

Background: The stability of mammalian serum and urine in large metabolomic investigations is essential for accurate, valid and reproducible studies. The stability of mammalian serum and urine, either processed immediately by freezing at -80 degrees C or stored at 4 degrees C for 24 h before being frozen, was compared in a pilot metabolomic study of samples from 40 separate healthy volunteers.

Methods: Metabolic profiling with GC-TOF-MS was performed for serum and urine samples collected from 40 volunteers and stored at -80 degrees C or 4 degrees C for 24 h before being frozen at -80 degrees C. Subsequent Wilcoxon rank sum test and Principal Components Analysis (PCA) methods were used to assess whether differences in the metabolomes were detected between samples stored at 4 degrees C for 0 or 24 h.

Results: More than 700 unique metabolite peaks were detected, with over 200 metabolite peaks detected in any one sample. PCA and Wilcoxon rank sum tests of serum and urine data showed as a general observation that the variance associated with the replicate analysis per sample (analytical variance) was of the same magnitude as the variance observed between samples stored at 4 degrees C for 0 or 24 h. From a functional point of view the metabolomic composition of the majority of samples did not change in a statistically significant manner when stored under two different conditions.

Conclusions: Based on this small pilot study, the UK Biobank sampling, transport and fractionation protocols are considered suitable to provide samples, which can produce scientifically robust and valid data in metabolomic studies.