This research was carried out to determine the effect of different hectoliter weights on some chemical properties of barley under laboratory conditions in 4 replications. In this study, barley having 4 different hectoliters weight (55, 60, 65 and 70 kg hl-1) was used and crude protein, crude fiber, crude oil, crude ash and starch content were examined. The datas obtained were subjected to variance analysis according to the randomized plot design. According to the analysis of variance, the hectoliter weight had a statistically significant effect on all characters at the level of 1%. According to the results, crude protein ranged 9.83-12.18%, crude fiber 4.14-6.26%, crude oil 1.43-1.51%, crude ash 2.40-2.98% and starch 47.63-53.17% and as the hectoliter weight increased, the crude protein and crude fiber and crude ash content decreased, while the crude oil and starch content increased.