Combined 1H-NMR and 1H–13C HSQC-NMR to improve urinary screening in autism spectrum disorders

Autism spectrum disorders (ASD) are neurodevelopmental diseases with complex genetic and environmental etiological factors. Although genetic causes play a significant part in the etiology of ASD, metabolic disturbances may also play a causal role or modulate the clinical features of ASD. The number of ASD studies involving metabolomics is increasing, and sometime with conflicting findings. We assessed the metabolomics profiling of urine samples to determine a comprehensive biochemical signature of ASD. Furthermore, to date no study has combined metabolic profiles obtained from different analytical techniques to distinguish patient with ASD from healthy individuals. We obtained 1H-NMR spectra and 2D 1H–13C HSQC NMR spectra from urine samples of patients with ASD or healthy controls. We analyzed these spectra by multivariate statistical data analysis. The OPLS-DA model obtained from 1H NMR spectra showed a good discrimination between ASD samples and non-ASD samples (R2Y(cum) = 0.70 and Q2 = 0.51). Combining the 1H NMR spectra and the 2D 1H–13C HSQC NMR spectra increased the overall quality and predictive value of the OPLS-DA model (R2Y(cum) = 0.84 and Q2 = 0.71), leading to a better sensitivity and specificity. Urinary excretion of succinate, glutamate and 3-methyl-histidine differed significantly between ASD and non-ASD samples. Urinary screening of children with neurodevelopmental disorders by combining NMR spectroscopies (1D and 2D) in multivariate analysis is a better sensitive and a straightforward method that could help the diagnosis ASD.