This paper reviews the up-to-date research about the applications of graphene and its related materials in the field of mass spectrometry (MS). Due to its large surface area, delocalized pi-electrons, thermal conductivity, stability and rich interaction chemistry, graphene has been widely used in MS-based analytical chemistry. Graphene-based materials were applied as very effective matrixes or surfaces for many kinds of organic molecules in laser desorption/ionization (LDI) MS analysis. Many advantages of this novel matrix have been proved, which included: low interference ions from matrix itself, good reproducibility, high salt tolerance and so on. The unique properties of graphene also make it a superior sorbent used in solid-phase extraction (SPE). Further development of online SPE methods based on graphene coupling directly with LDI-MS, GC-MS and LC-MS greatly simplifies the MS-based analytical procedure for complex samples and makes the corresponding high-throughput and automatic analysis performable. Their applications as a platform in proteolysis for the rapid identification of proteins have been also developed. In addition, graphene was found to be a unique precursor for the generation of large-sized carbon cluster anions in the gas phase. Finally, the possible challenges and future perspectives in their applications in MS are discussed too.