OBJECTIVE Galangin,apotent scavenger of free radicals,is used as herbal medicine for various ailments for centuries in Asia.With complex pathophysiology,ischemic stroke is one of the most frequent causes of death and disability worldwide.We have reported that galangin provides a direct protection against ischemic injury as a potential neuroprotective agent and has potential therapeutic effects on the changes of serum amino acids for ischemic stroke;however,its mechanism on changes of amino acids in the ischemic brain tissue has not yet been clarified.METHODS In this paper,we explored the amino acid biomarkers of brain tissue in the acute phase of cerebral ischemia and the effect of galangin on those potential biomarkers with a rapid,sensitive and accurate methodology of simultaneous quantification of 12 AAs in rat brain tissue by the RRLC/QQQ.RESULTS we identified that glutamic acid,alanine and aspartic acid all showed significant change in galangin-treated groups compared to vehicle-treated rats and four pathway-related enzymes were identified by multiplex interactions with the three amino acids.With metabolite-protein network analysis and molecule docking,six of 28 proteins were identified and may become the potential biomarkers of galangin for acute ischemic stroke.CONCLUSION All data in our study provide thought for exploring the mechanism of disease,discovering new targets for drug candidates and elucidating the related regulatory signal network. OBJECTIVE Galangin, apotent scavenger of free radicals, is used as herbal medicine for various ailments for centuries in Asia. Wl complex pathophysiology, ischemic stroke is one of the most frequent causes of death and disability worldwide. We have reported that galangin provides a direct protection against ischemic injury as a potential neuroprotective agent and has potential therapeutic effects on the changes of serum amino acids for ischemic stroke; however, its mechanism on changes of amino acids in the ischemic brain tissue has not been been clarified. METHHODS In this paper, we explored the amino acid biomarkers of brain tissue in the acute phase of cerebral ischemia and the effect of galangin on those potential biomarkers with a rapid, sensitive and accurate methodology of simultaneous quantification of 12 AAs in rat brain tissue by the RRLC / QQQ.RESULTS we identified that glutamic acid, alanine and aspartic acid all showed significant change in galangin-treated groups compared to vehicle-t reated rats and four pathway-related enzymes were identified by multiplex interactions with the three amino acids. Wheat metabolite-protein network analysis and molecule docking, six of 28 proteins were identified and may become the potential biomarkers of galangin for acute ischemic stroke. CONCLUSION All data in our study provide thought for exploring the mechanism of disease, discovering new targets for drug candidates and elucidating the related regulatory signal network.