Abstract: Eurotium cristatum had important application value in the development of hypoglycemic and lipid-lowering functional foods and natural medicines. The analysis of the overall metabolite information and the hypoglycemic and lipid-lowering active ingredients produced by Eurotium cristatum could provide theoretical basis and data support for the development and application of the bacterial strain. By taking the strain Ys-Ec-01 of Eurotium cristatum as the experimental material, the non-targeted metabolomics based on the ultra performance liquid chromatography-quadrupole electrostatic field orbitrap mass spectrometry (UHPLC-QE-MS) technology was used to analyze the fermentation metabolic profile Ys-Ec-01 by combining with the principal component analysis (PCA) and the orthogonal partial least squares-discriminant analysis (OPLS-DA), in order to find the hypoglycemic and lipid-lowering active products. The results showed that the metabolite profile of Ys-Ec-01 changed significantly after the fermentation. From the metabolite profile, a total of 365 fermentation products were identified, and then 205 differential fermentation products and 151 main fermentation products were screened, thus to identify 13 hypoglycemic and lipid-lowering active ingredients, which were chrysoeriol, kaempferol, isorhamnetin, epicatechin, capsaicin, nicotinic acid, trigonelline, 1-deoxynojirimycin, allantoin, protocatechuic acid, linoleic acid, choline and L-carnitine. These active ingredients provided more targets and basic information for the research, development and application of hypoglycemic and lipid-lowering function of Ys-Ec-01.