Assessment of 1H NMR-based metabolomics analysis for normalization of urinary metals against creatinine

Background: Proton nuclear magnetic resonance (H-1 NMR, or NMR) spectroscopy and inductively coupled plasma-mass spectrometry (ICP-MS) are commonly used for metabolomics and metal analysis in urine samples. However, creatinine quantification by NMR for the purpose of normalization of urinary metals has not been validated. We assessed the validity of using NMR analysis for creatinine quantification in human urine samples in order to allow normalization of urinary metal concentrations. Methods: NMR and ICP-MS techniques were used to measure metabolite and metal concentrations in urine samples from 10 healthy subjects. For metabolite analysis, two magnetic field strengths (600 and 700 MHz) were utilized. In addition, creatinine concentrations were determined by using the Jaffe method. Results: Creatinine levels were strongly correlated (R-2 = 0.99) between NMR and Jaffe methods. The NMR spectra were deconvoluted with a target database containing 151 metabolites that are present in urine. A total of 50 metabolites showed good correlation (R-2 = 0.7-1.0) at 600 and 700 MHz. Metal concentrations determined after NMR-measured creatinine normalization were comparable to previous reports. Conclusions: NMR analysis provided robust urinary creatinine quantification, and was sufficient for normalization of urinary metal concentrations. We found that NMR-measured creatinine-normalized urinary metal concentrations in our control subjects were similar to general population levels in Canada and the United Kingdom. (C) 2016 Elsevier B.V. All rights reserved.