This paper describes the first metabolomic analysis using proton high resolution nuclear magnetic resonance (1H HR NMR) spectroscopy in three patients with epileptic encephalopathy due to the recurrent c.320A>G variant in ALG13. The metabolome characteristics of these three female patients are presented, giving assumptions for further studies that could verify the role of three metabolites as biomarkers suitable for clinical or treatment assessment. This evidence concerns the gene ALG13 and Epileptic encephalopathy.