Cell extraction method coupled with LC-QTOF MS/MS analysis for predicting neuroprotective compounds from Polygonum tinctorium

The cell extraction method coupled with LC-QTOF-MS/MS is a biological screening technique in which cells are incubated with extracts of natural products, which results in potential bioactive compounds selectively combining with various extracellular and intracellular targets. Although the neuroprotective effects of the plant Polygonum tinctorium are unknown, the ethyl acetate (EtOAc) fraction exhibits significant neuroprotective effects against ʟ-glutamate-induced cytotoxicity in HT22 cells. In this study, we attempted to identify the neuroprotective compounds in the EtOAc fraction of P. tinctorium using the cell extraction method coupled with LC-QTOF MS/MS. Potential neuroprotective components derived from P. tinctorium were combined selectively with HT22 cells, and cell-derived metabolites were identified. A new flavonoid compound, 3,5,3',4'-tetrahydroxy-6,7-methylendioxyflavone-3-O-β-ᴅ-glucopyranoside (1), and 14 known compounds (2-15), with compounds 2, 3, 8, 13, and 15 detected by the cell extraction method, were isolated from the EtOAc fraction of P. tinctorium. Compounds 2, 8, 12, and 14 showed strong neuroprotective effects, with compounds 2 and 14 identified in this plant for the first time in this study. Our results indicate that the cell extraction method coupled with LC-QTOF MS/MS is a useful tool for screening and identifying neuroprotective compounds in natural products.