The extraction conditions of the tannin from mature cotton fiber were optimized,and the three factors including the ratio of extracting solution and material,the time and temperature of extraction were respectively carried out by single factor test.The optimum conditions of extraction were obtained as follows:the ratio of extracting solution and material was 30∶1mL/g,the extraction temperature was 85℃ and extraction time was 60min.The contents of tannin in cotton from different grades and producing area were determined under the optimum conditions,and the results showed that it had the large difference in contents of tannin in cotton fiber between different areas and different grades.