The aim is to establish a HPLC method for the determination of related substances in glucosamine hydrochloride capsules. The separation was performed on a C18(250mm*4.6mm, 5μm) column at room temperature The mobile phase was a mixture of sodium 1-heptanesulfonate solution and acetonitrile at a flow rate of 1.0mL·min-1. The detection wavelength was 195nm. The glucosamine hydrochloride capsule was stable with acid destruction, but unstable with alkali, high temperature and strong oxidative destruction. The linear range was 6.32-31.60μg·mL-1. This method was simple, accurate, and rapid with high specificity. It was applicable for the determination of related substances in glucosamine hydrochloride capsules.