title: Contribution towards a Metabolite Profile of the Detoxification of Benzoic Acid through Glycine Conjugation: An Intervention Study.
abstract: Benzoic acid is widely used as a preservative in food products and is detoxified in humans through glycine conjugation. Different viewpoints prevail on the physiological significance of the glycine conjugation reaction and concerns have been raised on potential public health consequences following uncontrolled benzoic acid ingestion. We performed a metabolomics study which used commercial benzoic acid containing flavored water as vehicle for designed interventions, and report here on the controlled consumption of the benzoic acid by 21 cases across 6 time points for a total of 126 time points. Metabolomics data from urinary samples analyzed by nuclear magnetic resonance spectroscopy were generated in a time-dependent cross-over study. We used ANOVA-simultaneous component analysis (ASCA), repeated measures analysis of variance (RM-ANOVA) and unfolded principal component analysis (unfolded PCA) to supplement conventional statistical methods to uncover fully the metabolic perturbations due to the xenobiotic intervention, encapsulated in the metabolomics tensor (three-dimensional matrices having cases, spectral areas and time as axes). Identification of the biologically important metabolites by the novel combination of statistical methods proved the power of this approach for metabolomics studies having complex data structures in general. The study disclosed a high degree of inter-individual variation in detoxification of the xenobiotic and revealed metabolic information, indicating that detoxification of benzoic acid through glycine conjugation to hippuric acid does not indicate glycine depletion, but is supplemented by ample glycine regeneration. The observations lend support to the view of maintenance of glycine homeostasis during detoxification. The study indicates also that time-dependent metabolomics investigations, using designed interventions, provide a way of interpreting the variation induced by the different factors of a designed experiment-an approach with potential to advance significantly our understanding of normal and pathophysiological perturbations of endogenous or exogenous origin.
authors.affiliation: Centre for Human Metabolomics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.
authors.forename: Cindy
authors.initials: C
authors.lastname: Irwin
authors.affiliation: Centre for Human Metabolomics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.|Department of Statistics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.
authors.forename: Mari
authors.initials: M
authors.lastname: van Reenen
authors.affiliation: Centre for Human Metabolomics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.
authors.forename: Shayne
authors.initials: S
authors.lastname: Mason
authors.affiliation: Centre for Human Metabolomics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.
authors.forename: Lodewyk J
authors.initials: LJ
authors.lastname: Mienie
authors.affiliation: Department of Statistics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.|Biosystems Data Analysis, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands.
authors.forename: Johan A
authors.initials: JA
authors.lastname: Westerhuis
authors.affiliation: Centre for Human Metabolomics, Faculty of Natural Sciences, North-West University (Potchefstroom Campus), Potchefstroom, South Africa.
authors.forename: Carolus J
authors.initials: CJ
authors.lastname: Reinecke
mesh_terms: D000328:Adult; D019817:Benzoic Acid / Q000378:metabolism*; D016000:Cluster Analysis; D005998:Glycine / Q000378:metabolism*; D064368:Healthy Volunteers; D006801:Humans; D008658:Inactivation, Metabolic; D008297:Male; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D055432:Metabolomics / Q000379:methods; D066244:Proton Magnetic Resonance Spectroscopy; D013997:Time Factors; D055815:Young Adult
chemical_list: D019817:Benzoic Acid; D005998:Glycine