The extraction and separation of Alkaloids in seeds of Sophora alopecuroides were studied in the experiment, the quantitative and qualitative analysis in matrine were also detected. Alkaloids were extracted from seeds by immersing in 0.4% hydrochloric acid, 95% alcohol, 1% ammonia-chloroform and anhydrous alcohol for four times in seven days under the room temperature after crushed, accompanying by delipoid, acidification, alkalization, extraction. Single Alkaloids were separated by TLC method, and quantitative and qualitative analysis was done by GC method. The results suggested that extraction yield of four methods above were 3.67%,2.73%,2.47% and 1.60% respectively. The extracted Alkaloids in each method contained four kinds of single Alkaloids at least by TLC. The extraction Alkaloids of all these four methods contained Kushen Alkaloids detected by GC, and the extraction yield were 8.460%、7.553%、5.675%and0.893% respectively. The results showed that extraction yield by 0.4% hydrochloric acid under room temperature was obviously higher than other solvents, which could be the optimal extraction method of Alkaloids in seeds of Sophora alopecuroides.