The premise and key of ensuring the safety and effectiveness of traditional Chinese medicine (TCM) is to construct appropriate quality evaluation system of TCM. This study aimed to establish a pre-column derivatization HPLC method for achieving the quality control of by reacting synthesized 4-hydrazino-1,8-naphthalimide (HAN) with diverse monosaccharides from the hydrolytic product of polysaccharides (PSPs), followed by HPLC separation. The HAN was synthesized based on a CuI-catalyzed cross-coupling reaction in water, and then employed as a novel chemosensor that reacts with reducing sugars. Good separation was achieved at a detection wavelength of 448 nm using an ZORBAX SB-C8 column under a gradient elution at a flow rate of 0.5 ml/min within 12 min. The monosaccharide compositions of PSP mainly include two hexoses [glucose (Glc), galactose (Gal)] and two hexuronic acids [glucuronic acid (GlcA) and galacturonic acid (GalA)], and the molar ratio of Glc, Gal, GlcA and GalA is 16.67:52.94:10.58:19.81. The verified HPLC method, possessing excellent precision and good accuracy, successfully achieved rapid qualitative and quantitative determination for PSP Additionally, the HAN displayed fluorescence enhancement through "push-pull" mode, and fluorescence decreased through "pull-pull" mode after binding to monosaccharides, which is a potential for fluorescence determination of different monosaccharides.