Objective: Ginkgo biloba leaves, as an herbal medicine or dietary supplement, have been widely used worldwide. In this study, an integrated analytical method was established for the comprehensive analysis of flavonoids in G. biloba leaves. Materials and Methods: A practical chromatographic method combining high?performance liquid chromatography fingerprint analysis and quantitation was used to simultaneously determine 11 flavonoids (6 flavonol glycosides and 5 biflavones) in G. biloba leaves from different regions. Results: A total of 11 characteristic peaks were identified accurately, and the similarity of fingerprints ranged from 0.944 to 0.996. Methodology validation revealed appropriate linearity (R2 ≥0.9997), precision, repeatability, stability, and recovery. The total contents of the six flavonol glycosides and five biflavones were within the range of 2.142-8.378 mg/g and 3.759-5.675 mg/g in 19 batches of samples, respectively. Among them, two coumaroyl flavonol glycosides were the predominant components. Conclusions: In this study, a convenient and reliable approach was successfully employed for the comprehensive evaluation of flavonoids in G. biloba leaves, which also provided a reference for its quality standard.