The invention provides a high-performance liquid chromatography (HPLC) method for simultaneously determining 16 ginseng saponin monomers, such as Rg1, Re, Rf, Rb1, Rg2, Rc, Rb2, Rb3, F1, Rd, F2, Rg3, protopanaxatriol, Compound K, Rh2 and protopanaxadiol. The determination range comprises medicinal plants such as ginseng and American ginseng containing more than one saponin monomer, and preparations thereof. Compared with the prior art, the HPLC method has the advantages that the variety of the ginseng saponin monomers simultaneously determined by the method can be up to 16, and the efficiency is improved. Only are acetonitrile and water used as mobile phases, so that the problems that phosphate is large in loss on a chromatographic column, the system is troublesome to clean and the like are solved. The built HPLC method is wide in linearity range, accurate in method, good in reproducibility, fast and convenient, stable and reliable. A theoretical basis is provided for quality control of the medicinal plants, such as ginseng and American ginseng, and the preparations thereof.