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Mass spectrometry is a sophisticated analytical technology capable of precisely measuring the mass-to-charge ratio of ions, which allows researchers to calculate ion mass at the atomic level with exceptional accuracy. The hyphenated technique of chromatography coupled with mass spectrometry has become an indispensable and powerful analytical tool. It is widely applied to identify and quantify target compounds and their metabolites in biological specimens and pharmacokinetic research, as well as to screen and monitor pollutants present in food and environmental samples. This Topic covers a full range of chromatographic techniques including gas chromatography, high-performance liquid chromatography, thin-layer chromatography, and supercritical fluid chromatography, and also includes various combined analytical technologies such as GC-MS, LC-MS/MS, ICP-MS, LC-NMR, and two-dimensional chromatography, aiming to present cutting-edge achievements and promote the further development and application of related analytical technologies.