Objective: In this study, a comprehensive and effective quality method for evaluating the efficacy of ShuXueNing injection (SXNI) was developed. Materials and Methods: Quantitative high?performance liquid chromatography fingerprint, the quantitative analysis of multicomponents by a single marker (QAMS) method, hierarchical cluster analysis (HCA) , and orthogonal partial least squares discrimination analysis (OPLS?DA) were used to distinguish 53 batches of SXNI samples from 7 manufacturers. Results: A total of 53 batches of samples were analyzed to establish antithesis fingerprint of SXNI, and 12 peaks of the common model were collected and used for the similarity analysis. Meanwhile, six index flavonoid components were determined by the QAMS method, using rutin as internal reference substance. The accuracy of the QAMS method was confirmed by investigating the relative deviation between the QAMS method and the traditional external standard method. The results demonstrated that there was no significant difference (RE < 1％), suggesting that QAMS was a reliable and convenient method for the content determination of multiple components. The HCA and OPLS?DA methods drew a similar conclusion. The 53 batches of SXNI samples from 7 manufacturers were categorized into five groups, indicating that chemometrics could reveal the quality differences of SXNI between the manufacturers. Conclusions: The method established herein was efficient and successful in assessing the quality of SXNI, and that it may be potentially employed in the quality control of related products composed of Ginkgo biloba extract.