Use of UHPLC-QTOF-MS/MS with combination of in silico approach for distributions and metabolites profile of flavonoids after oral administration of Niuhuang Shangqing tablets in rats

Niuhuang Shangqing tablet (NHSQT), a well-known traditional Chinese medicine preparation, has been used as an over- the- counter drug for the treatment of headache, dizziness in China. The flavonoids are the main active components in NHSQT, however, there have no reports about their distribution and metabolic fate in vivo after oral administration of NHSQTs so far. An novel UHPLC-QTOF-MS/MS method combined with in silica approach was applied to identify the flavonoids and metabolites profiling in biological samples following oral administration NHSQTs for the first time. As a result, 127 compounds including 34 original compounds of flavonoids and 93 metabolites were identified. There were 20 flavones, 9 flavonols, 4 flavanones and 1 flavan-3, 4-diol found in biological samples. Rutin, wogonoside, apigenin, baicalein, wogonin, oroxylin A, quercetin and acacetin were considered as the potential flavonoids in NHSQT against brain diseases. The docking-based metabolism models were established and applied to propose the sites of hydroxylation of flavonoids, which indicated baicalin was engaged in dihydroxylation at C2', C3', tilianin was engaged in hydroxylation at C3, wogonin and wogonside were engaged in dihydroxylation at C3', C4'. Some novel metabolic pathways were discovered for oroxylin A, acacetin, diosmetin, tilianin. The metabolic spots and pathways of flavonoids vary as much between flavones, flavonols and flavanones. The results presented here would be helpful for the further study of pharmacokinetics and quality control of NHSQT.