The invention relates to a large-volume sample injection-variable temperature focusing supercritical fluid chromatography mass spectrometry analysis system. The system comprises a modifier pump, a carbon dioxide pump, a mixer, an automatic sample injector, a six-way valve for switching flow paths, a quantitative loop, a column oven, a trace preheater, a chromatographic column, a supplement pump, a three-way connector, a back pressure regulator, a mass spectrum detector and a connecting pipeline for connecting all the components, wherein the mixer, the trace preheater and the chromatographic column are arranged in the column oven. The invention also relates to an analysis method using the system. The analysis method comprises the following steps of (1) changing temperature; (2) loading a sample by an automatic sample injector; (3) performing large-volume sample injection-column head focusing; and (4) performing chromatographic separation and mass spectrometry. According to the method, a problem of solvent incompatibility in supercritical fluid chromatography is solved, so a problem of chromatographic peak broadening caused by large-volume sample introduction is avoided, the chromatographic peak shape of a compound is improved, and analysis sensitivity is improved.