High-Coverage Metabolome Analysis Reveals Significant Diet Effects of Autoclaved and Irradiated Feed on Mouse Fecal and Urine Metabolomics

Mol Nutr Food Res. 2021 Jun;65(12):e2100110. doi: 10.1002/mnfr.202100110. Epub 2021 May 10.


Scope: Using metabolomics to study the relations of nutrition and health requires stringent control of the experimental conditions used in an animal model. This work investigates the diet effects of autoclaved and irradiated feed on mouse urine and fecal metabolomics.

Methods and results: C57BL/6 mice are fed normal-irradiation sterilized diet (n = 9), autoclave sterilized diet (n = 9), and high-irradiation sterilized diet (n = 9) for 4 weeks. Differential chemical isotope labeling liquid chromatography mass spectrometry is used to quantify the metabolome variations of urine and feces collected at five time points. Significant differences are observed in urine or fecal metabolomes of mice fed autoclaved diet versus mice fed high-irradiation diet or fed normal-irradiation diet, while the differences are small between the mice fed normal-irradiation and high-irradiation diet. Correlation studies of metabolite changes of diet- and aging-related biomarkers indicate a large overlap of significantly affected metabolites by the two factors.

Conclusions: Diet can be a confounding factor that needs to be carefully considered when a metabolomics study is designed and metabolomic results of a mouse model of nutritional or other biological study are interpreted. Using the same sterilized diet for a given metabolomics project is essential to control the diet effect.

Keywords: LC-MS; diet effects; metabolomics; mouse feed; urine and feces.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aging / physiology
  • Animal Feed*
  • Animals
  • Body Weight
  • Chromatography, Liquid
  • Diet
  • Feces / chemistry*
  • Food Irradiation*
  • Mass Spectrometry / methods
  • Metabolome
  • Metabolomics / methods*
  • Mice, Inbred C57BL
  • Urinalysis / methods
  • Urine / chemistry*