Abstract: The G-quadruplex is an atypical secondary structure of nucleic acid formed by guanine-rich sequences, which exists in a variety of biological systems and plays an important role in regulating gene expression. So it has become a potential drug therapy target. Mass spectrometry technology is widely used in the study of G-quadruplexes because of its high sensitivity, high accuracy and low sample consumption. It has become a powerful tool for studying the structures of Gquadruplexes and the binding of G-quadruplexes with small molecule ligands. Mass spectrometry detection of G-quadruplex formation, binding affinity and stoichiometry, stabilization and collision dissociation behavior of G-quadruplex DNA, progress of MS research on RNA G-quadruplex, and other mass spectrometric techniques for conformational analysis of G-quadruplex would be reviewed in this study.