The invention discloses a method for determining the content of calcium oxalate in tobacco. According to the method, an acetic acid solution and a hydrochloric acid solution are used respectively for extracting a reconstituted tobacco sample, then the content of calcium elements in extracting solutions is determined by an ion chromatography, and the content of the calcium oxalate in the tobacco is calculated by using the difference of the content of calcium elements in the extracting solutions. To be specific, the method comprises the steps as follows: (1), grinding the tobacco sample into powder, filtering the sample by a sieve, determining the moisture, and reserving the sample for use; (2), weighing a certain quantity of the powder-like sample obtained in the step (1), using the acetic acid solution for extraction, after constant-temperature vibrating extraction for a certain period, performing centrifugation, taking out a supernatant, and determining the volume to obtain a sample A; (3), weighing another quantity of the powder-like sample obtained in the step (1), using the hydrochloric acid solution for extraction, after constant-temperature vibrating extraction for a certain period, performing centrifugation, taking out a supernatant, and determining the volume to obtain a sample B; and (4), determining the content of calcium element in the sample A and the content of calcium element in the sample B by the ion chromatography and calculating the content of the calcium oxalate by using the difference between the content. The method has the advantages of convenience in operation and high accuracy and stability.