The high performance gel permeation chromatographic (HPGPC) behaviors of four beta-1, 3/1, 6-glucans (GF1, GF2, GF3, GF4) with different protein contents and one alpha-1, 4/1, 6-glucan (P100) from Grifola frondosa were examined with different concentrations of NaCl solution and pH values. The experimental results showed that the relative molecular mass (M(r)) of the beta-glucans sharply dropped as the NaCl concentration was less than 0.025 mol/L and then tended to be stable as the NaCl concentration was raised from 0.1 to 0.2 mol/L, the M(r) increased quickly from pH 3 to 6, and then maintained stable from pH 6 to 9, but slightly increased as the pH value was higher than 9. However, the M(r) of the alpha-1, 4/1, 6-glucan was slightly affected by either different concentrations of NaCl or different pH values. The beta-glucan existed in a super-helix structure in aqueous solution, which could be influenced by the NaCl concentration and pH value. These factors led to the different molecular aggregation states, and the increase or decrease of the M(r) of beta-glucans, displaying a variety of HPGPC behaviors.