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Ganglioside and sulfatide are primary components of acidic glycosphingolipids (AGSLs), which are abundant in the brain tissue. AGSLs are potential tumor markers. In this paper, we use ultra-high performance liquid chromatography–quadrupole time-of-flight mass spectrometry to generate a first-ever comprehensive profile of AGSLs in brain and plasma of C6 glioma rats treated with temozolomide (TMZ). The AGSLs detected consisted mainly of C18-/C20-sphingosine. 12, 20, 19, 14 and 12 species were identified in GM1, GD1a, GD1b, GM3 and GT1b ganglioside groups which were abundant in rat brain, respectively. These five groups accounted for 88–89% of total ganglioside content. However, no AGSLs were detected in rat plasma. Possible biomarkers for abnormal changes in the glioma model and the protective effect of TMZ were mainly found in these ganglioside groups and sulfatide. The main lipid …