Summary: To explore the effects of wet milling method on highland barley flour and highland barley bread, we investigated the characteristics of highland barley flours and the quality of highland barley breads made from whole grain highland barley flour (WGBF), dry‐milled highland barley flour (DBF) and wet‐milled highland barley flour (WBF). Compared to other highland barley flours, WBF had lower damaged starch content (1.37 g/100 g), higher pasting temperature and higher gel strength. In addition, WBF with large particles exhibited low hydration performance. These properties induced a large specific volume (2.62 mL g−1) and good crumb structure of bread with WBF. However, due to the negative effects of damaged starch and insoluble dietary fibre in WGBF and DBF, the specific volumes of bread with WGBF and DBF (1.30 and 1.70 mL g−1) were lower than the bread with WBF. The results showed that WBF with high starch granules integrity could produce preferable highland barley bread. [ABSTRACT FROM AUTHOR]