Abstract: In the present study, we established an ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-QTOF-MSE) method to simultaneously quantify 33 components in Ginkgo biloba leaf extracts (GBEs), including 17 flavonol glycosides, five terpene trilactones (TTLs), four polyphenols and seven carboxylic acids. This optimized method was successfully applied to analyze the explicit compositions of GBE samples collected from different places. Furthermore, the data were processed through unsupervised principal component analysis (PCA) and supervised orthogonal partial least squared discrimination analysis (OPLS-DA) to evaluate the quality and compare the differences between the samples according to the contents of the 33 chemical constituents. Bilobalide, protocatechuic acid, shikimic acid, quinic acid, ginkgolide B, ginkgolide J, kaempferol-3-O-rutinoside, isorhamnetin-3-O-rutinoside, quercetin-3-O-ct-L-rhamnopyranocyl-2"-(6'"-p-coumaroyl)-β-D-glucoside and rutin were recognized as characteristic chemical markers that contributed most to control the quality of GBEs. Based on the fact that GBEs should be standardized with the characteristic components as quality control chemical markers, it is most important to maintain the quality of GBEs stable and reliable, and this method also provided a good strategy to further rectify and standardize the GBEs market.