The results of the comparative study on microbiological and chemical quantitative determination of kanamycin sulfate in the ophthalmic films with the collagen base are presented. The intraocular films prepared with the use of 1 per cent collagen solution contain dexamethasone and kanamycin. The agar diffusion method with Bacillus pumilus NCTC 8241 as the test microbe and the photocolorimetric method based on estimation of the optical density of the colored compound formed after acid hydrolysis of kanamycin with orcinol and ferric chloride were used for the quantitative determination of kanamycin. The results of the quantitative determinations of kanamycin in the films with the two methods did not differ significantly. However, the error of the microbiological method was +/- 3,75 per cent, whereas that of the chemical method was +/- 1.23 per cent or approximately 3 times lower. The time of the analysis decreased from 24 to 1.5-2 h. Moreover, the chemical method is simple and readily reproducible.