Atrophic vaginitis (AV) is a common and frequently occurring disease, lacking effective curative measures. Exploring the mechanism of vaginal mucosal homeostasis from the perspective of metabolites has great research prospects.We compared the metabolic profiles of vaginal secretions between AV patients and healthy individuals via liquid chromatography-tandem mass spectrometry (LC-MS/MS). We further explored effective and sensitive metabolites and metabolic pathways for senile vaginitis through bioinformatics analysis and experimental verification. Through untargeted metabolomics analysis, we screened 561 differential metabolites in two groups of vaginal secretion samples. These differential metabolites were mainly concentrated in fatty acids/carboxylic acids, glycerophospholipids, organic oxides, steroids, and their derivatives. They were mainly enriched in purine metabolism, diabetic cardiomyopathy generation, and choline metabolism pathways.The receiver operating characteristic analysis showed the metabolites (e.g., guggulsterone, umbelliprenin, and inosinic acid) to have good discrimination ability for the AV group. In addition, we also explored the potential mechanism of action of umbelliprenin at the cellular level.This study is expected to provide a new perspective for understanding the relationship between metabolites and the pathogenesis of AV.Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.