Due to its high sensitivity and selectivity, mass spectrometry (MS) is considered today as one of the most versatile and powerful detection methods. This detector is considered as the gold standard for analytical separations dealing with complex mixtures and biological matrices. Since the first report of supercritical fluid chromatography-mass spectrometry (SFC-MS) hyphenation in the 1980s, various ionization sources, mass analyzers, and interfacing technologies have been employed. The goal of the present chapter is to summarize all the possible combinations for successful SFC-MS operation and to highlight the most promising ones. Due to the physical properties and compressibility of SFC mobile phase, moving column effluent to the ionization source is less straightforward than in LC. Therefore specially designed interfaces have to be employed to limit the risk of analyte precipitation caused by the decompression of CO2 and to achieve sufficient ionization yield. As reported several commercial SFC-MS interfaces now allow robust and highly sensitive SFC-MS operation. SFC-MS generally offers better sensitivity, reduced matrix effects, and lower impact of MS operating conditions on sensitivity (enhanced robustness) versus LC-MS. This behavior was explained by the absence of water in the mobile phase and the fact that CO2 can be eliminated easily in the ionization source. In addition, SFC-MS has higher kinetic performance, is greener, more universal, and orthogonal analytical technique in comparison to LC-MS. As reported here, SFC-MS(/MS) was successfully employed in various fields including lipidomics, metabolomics, bioanalysis, food, and environmental analysis.