Objective To determine the content of polysaccharides and polypeptides in the 35% ethanol-water elution part of the effective part of ginkgo kernels, and to provide scientific basis for the comprehensive development and quality control of ginkgo kernels. Methods The standard curve was established by the phenol-sulfuric acid method and biuret method, and the precision, stability, repeatability, and recovery rate were investigated. Results The polysaccharides and polypeptides in the 35% ethanol-water elution part of ginkgo kernel had a good linear relationship in the concentration range. The RSD values of methodological verification were less than 2%. The recovery rates were 97.96 % and 97.94 %. The polysaccharides content was 13.99%, and the polypeptides content was 31.41%. Conclusion The established method for the determination of polypeptides and polysaccharides in the 35% ethanol-water elution part of ginkgo kernels is simple and feasible, with good stability and reproducibility.