This study was aimed to determine metabolites produced by Magnaporthe oryzae and identify metabolic changes during appressorium development. Appressorium development were induced in vitro and subjected to 1H NMR spectroscopy for metabolites production and multivariate data analysis. PCA, PLS-DA and OPLS-DA were used to profile metabolite production throughout appressorium development. There were 43 metabolites identified putatively and PCA showed differences of metabolites production between mycelium and appressorium development. Metabolites that were significantly produced (p < 0.05) during appressorium development including isocitrate, isobutyrate, lysine, glutamate, succinate, tyrosine, choline, glycerol, xylose, mannose, sucrose, tryptophan, butyrate, leucine, isoleucine, valine, ethanol, methylmalonate, threonine, lactate, alanine, arginine, 4-aminobutyrate, homoserine, glucose, mannitol and glucitol. Glycerolipid, carbohydrates and amino acids metabolisms showed to be highly involved during appressoria development. This study revealed metabolites produced by M. oryzae during appressoria development in vitro as first metabolomics data using 1H NMR approach.