A higher biopotency of 26053.93 IU/mg was obtained from the urine of pregnant women using different purification methods. The bioassay of hCG activity was tested on rats based on rat seminal vesicles weight gain. Pregnant women urine was extracted and purified with ion exchange chromatography and were purified from 50 IU to 1500 IU with protein content from 243 mg to 8.12 mg. As a consequence of these results, 4.23 mg of protein content was obtained with a biological activity of 3000 IU/mg. To attain increased purity of HCG, the product was subjected and allowed to pass through the affinity column chromatography. This led to the formation of the highly purified HCG with >26000 IU/mg of biological activity.