The invention provides a kit and method for rapidly detecting the endocrine disrupting activity of a chemical substance based on magnetic microspheres. The kit comprises a microsphere-GST/6*HIS-NR LBDprotein polymer solution, an SRC-BAP protein solution, a BSA Solution, a reaction solution, a washing solution, a color developing solution and a coloring stop solution. According to the invention, the activated microsphere-protein polymer solution, the SRC-BAP protein solution, the BSA solution and a to-be-detected substance are mixed and incubation is performed; after cleaning, the color developing solution is added and color development is carried out and then is stopped; and a light absorption value is measured by a common optical microplate reader; the light absorption value is processedby software and EC50 or IC50 is calculated, so that the receptor activity of a chemical substance is quickly determined. With the provided detecting method, the experimental period can be shortened,the purity and activity of the receptor can be enhanced, and the detection sensitivity can be improved, so that an objective of detecting and screening the endocrine disrupting receptor activity in the chemical product sensitively, efficiently, and rapidly is achieved.