Arabinoxylan (AX) content in barley grains is an important quality determinant when barley is used as raw material of malt or beer production. The cultivar and environmental variations of total arabinoxylan (TAX), water extractable arabinoxylan (WEAX) and endoxylanase activity (EA) were investigated using eight barley cultivars growing at seven locations with diverse environmental conditions. The results showed that both barley cultivar and location significantly affected the TAX, WEAX and EA levels, but the variations of TAX content and EA were mainly attributed to cultivar, while the impact of location on WEAX content was greater than that of cultivar. Correlation analysis indicated that TAX was significantly correlated to WUAX.