5-hydroxymethylfurfural (5-HMF) is a kind of new green platform chemical with wide application. Glucose, which is the unit compound of cellulose, is one of the most important starting chemicals from biomass. With its low cost and wide supply, the conversion of glucose to HMF has attracted the interests of researchers. In this work, a systematic study has been conducted to evaluate the catalytic performance in the conversion of glucose to 5-HMF using Zr(H2PO4)2 solid acid catalyst in isobutanol-water (1.6:1/V:V) two-phase system. An optimized 5-hydroxymethylfurfural yield of 42.32% was obtained within 1.5 h at 180 °C over Zr(H2PO4)2 with ratio of 1:1 and the glucose conversion was 94.83%. The catalyst stabilities was studied and results showed that Zr(H2PO4)2 had higher stabilities.