Abstract: Anthocyanin, as one of the major bioactive components in tea, represents the edible value. Due to the enrichment of tea in protein, sugar, tea polyphenols, flavonoids, alkaloids, minerals, vitamins, pigments and other components, it is difficult to determine the anthocyanin active substances in tea matrix by relevant domestic standards. Thus, this paper established a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the detection of six anthocyanin compounds in tea sample under the optimization of pretreatment conditions and instrument conditions. The established method showed a good linear relationship within the concentration range of 25 to 2500 μg/L, with correlation coefficients (R) greater than 0.99, recovery rates ranging from 91.95% to 106.99%, and precision RSD less than 10%, which meet the requirements for analytical performance. The method exhibited advantages such as easy operation, high sensitivity, and high accuracy. These advantages make it suitable for related analytical detections and providing valuable guidance for the establishment of standard methods for anthocyanin content detection in tea.