Five winter wheat cultivars from the north and south wheat areas were selected as experimental materials. Through the different sowing date test, the amino acid quality of winter wheat grain was evaluated by variance analysis, principal component analysis and cluster analysis. Based on typical correlation and stepwise regression analysis, the correlation between winter wheat amino acid quality and climatic-biological factors was studied, and significant relevant climatic ecological factors were selected to construct winter wheat amino acid content prediction model. The results show that the average content of glutamate (non-essential amino) was the highest in amino acid contents of winter wheat, and the average content of methionine (essential amino acid) was the lowest. The environmental adaptability of essential amino acids and non-essential amino acids was relatively strong, while semi-essential amino acids had a greater potential for quality breeding. The content of amino acid in winter wheat showed the regional distribution characteristics that for northern varieties it was higher than that of southern varieties. The regional differences of threonine, phenylalanine, arginine, aspartic acid, glutamic acid and glycine were significant. The amino acid quality could be explained by three principal components with a cumulative contribution rate of 97.796%. In the comprehensive quality evaluation, the Gucheng Tanmai 98, and Taian Shannong 18 performed better, while the Jingzhou Zhengmai 9023, Suzhou Wanmai 52, Xuzhou Xumai 33 performed poorly. The cluster analysis showed that the cluster arrangement was closely related to the content of amino acid components and geographical distribution of winter wheat. The types I, II, III and IV were the Tanmai 98 in the north wheat area, the Shannong 18 in the northern part of Huanghuai wheat area, the Wanmai 52 in the southern part of Huanghuai, the Xumai 33 in the southern wheat area of Huanghuai and the Zhengmai 9023 in the Jianghuai wheat area, respectively which was consistent with the comprehensive evaluation ranking in the principal component analysis. The correlation between amino acid content of winter wheat and climatic-biological factors was obvious. The essential amino acid content was most correlated with temperature and humidity conditions. Most of the amino acid components could be improved in quality by adjusting the microclimate environment or soil moisture.