IntroductionGinkgo biloba extract (GBE) is an effective substance from traditional Chinese medicine (TCM) G. biloba for treating ischaemic stroke (IS). However, its active ingredients and mechanism of action remain unclear.ObjectivesThis study aimed to reveal the potential active component group and possible anti‐IS mechanism of GBE.Materials and methodsThe network pharmacology method was used to reveal the possible anti‐IS mechanism of these active ingredients in GBE. An ultra‐high‐performance liquid chromatography triple quadrupole electrospray tandem mass spectrometry (UPLC‐MS/MS) method was established for the simultaneous detection of the active ingredients of GBE.ResultsThe active components of GBE anti‐IS were screened by literature integration. Network pharmacology results showed that the anti‐IS effect of GBE is achieved through key active components such as protocatechuic acid, bilobalide, ginkgolide A, and so on. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis showed that the possible anti‐IS mechanism of GBE is regulating the PI3K‐Akt signalling pathway and other signal pathways closely related to inflammatory response and apoptosis regulation combined with AKT1, MAPK, TNF, ALB, CASP3, and other protein targets. Nineteen main constituents in seven batches of GBE were successfully analysed using the established UPLC‐MS/MS method, and the results showed that the content of protocatechuic acid, gallic acid, ginkgolide A, and so forth was relatively high, which was consistent with network pharmacology results, indicating that these ingredients may be the key active anti‐IS ingredients of GBE.ConclusionThis study revealed the key active components and the anti‐IS mechanism of GBE. It also provided a simple and sensitive method for the quality control of related preparations.