The invention discloses a purification method of beta-glucuronidase. The method comprises the steps of taking adult snails, cutting off shells, taking down soft parts, cutting off body walls in a culture dish, pulling out digestive glands, and putting digestive glands into a culture dish; puncturing the digestive glands by using a needle head, collecting the flowing viscous liquid and the digestive glands respectively, centrifuging under a certain condition to remove precipitates, combining supernate, and filtering by using a filter membrane to obtain a crude extract containing beta-glucuronidase; enabling the crude extract to pass through a DEAE ion exchange column for linear gradient elution, firstly determining the enzyme activity of the obtained eluents, then combining the eluents withrelatively high enzyme activity to obtain a beta-glucuronidase purified product; adding a preservative and an antioxidant; and performing low-temperature preservation. According to the invention, thepurification method is simple and feasible; the active material (snail) is rich in source, low in price, easy to obtain and short in period; and the purified beta-glucuronidase is high in activity, good in stability and high in recovery rate, can completely meet the requirements of an in-vitro diagnostic kit, and has relatively high development and application values.