Gliomas are tumors originating from the glial cells, accounting for ~28% of all CNS tumors. They are characterized by rapid proliferation rate and poor prognosis. Around 80% of the gliomas are malignant with a median survival of about 10-12 months. Based on the immunohistology, microscopic properties and mutational status, gliomas are classified into four grades. The current study was performed to develop MRM assays for validating candidate proteins arising from previous shotgun proteomics experiment and holds potential to distinguish between different grades of gliomas.