Library-based UHPLC-Q-exactive-orbitrap-MS putative identification of isomeric and non-isomeric bioactives from Zibushengfa Tablet and pharmacopeia quality-marker chemistry

Abstract Zibushengfa Tablet is a widely consumed traditional Chinese medicine (TCM) prescription. However, its main bioactives remain unclear and its pharmacopeia quality-marker (Q-marker) is poorly characterized nowadays. The present study analyzed its lyophilized aqueous powder using ultra-high-performance liquid chromatography-quadrupole-orbitrap mass spectrometry. Through comparison with authentic standards in the library, 46 bioactives were putatively identified, including quinic acid, D-gluconic acid, gallic acid, protocatechuic acid, 5-caffeoylquinic acid, L-tryptophan, salidroside, (+)-catechin hydrate, caffeic acid, cryptochlorogenic acid, mangiferin, ferulic acid, myricetin-3-O-galactoside, 2,3,5,4’-tetrahydroxystilbene-2-O-glucoside, forsythoside B, myricitrin, rutin, hyperoside, acteoside, 3,4-dicaffeoylquinic acid, myricetin, senkyunolide A, isochlorogenic acid C, quercitrin, (+)-epipinoresinol, quercetin, biochanin A, calycosin, luteolin, wedelolactone, calycosin-7-O-glucoside, chrysoeriol, trans-cinnamic acid, formononetin, chrysin, 3,3’,4’,5,6,7,8-heptamethoxyflavone, Z-ligustilide, amentoflavone, tangeretin, hinokiflavone, emodin, physcion, linoleic acid, stearic acid, palmitic acid, and palmitic acid ethyl ester. Of these, four pairs of isomers were successfully differentiated using the newly established library-filtering strategy. On this basis, senkyunolide A, amentoflavone, quercetin, quercitrin, and wedelolactone are preliminarily suggested as additional quality-markers for the consideration by Pharmacopeia Committee, to prevent the possible adulterate. Graphical Abstract