Due to the similarity in the grain and difference in the market value among many rice varieties, deliberate mislabeling and adulteration has become a serious problem. To check the authenticity, we aimed to discriminate rice varieties based on their volatile organic compounds (VOCs) composition by headspace solid phase microextraction (HS-SPME) coupled with gas chromatography mass spectrometry (GC–MS). The VOC profiles of Wuyoudao 4 from nine sites in Wuchang were compared to 11 rice cultivar from other regions. Multivariate analysis and unsupervised clustering showed an unambiguous distinction between Wuchang rice and non-Wuchang rice. Partial least squares discriminant analysis (PLS-DA) demonstrated a goodness of fit of 0.90 and a goodness of prediction of 0.85. The discriminating ability of volatile compounds is also supported by Random forest analysis. Our data revealed eight biomarkers including 2-acetyl-1-pyrroline (2-AP) that can be used for variation identification. Taken together, the current method can readily distinguish Wuchang rice from other varieties which it holds great potential in checking the authenticity of rice.