Identification of the constituents and metabolites in rats after oral administration of Zi Shen Formula by UPLC‐Q‐TOF/MS combined pattern recognition analysis

Abstract

An ultra-high-performance liquid chromatography mass spectrometry method was established to detect and identify the chemical constituents of Zi Shen Formula (ZSF) and its metabolites in serum, urine and feces, after oral administration to rats. A total of 68 compounds were characterized in ZSF extracts. In vivo, 38 prototype components and 32 metabolites of ZSF were tentatively identified in rat serum, urine and feces. Seven metabolic pathways including demethylation, hydroxylation, oxidation, sulfation, glucuronidation, methylation and de-caffeoyl were proposed to be involved in the generation of these metabolites. It was found that glucuronidation, methylation and demethylation were the major metabolic processes of alkaloids, while demethylation, methylation, sulfation and de-caffeoyl were the major metabolic pathways of phenylethanoid glycosides. The main metabolic pathways of steroidal saponins were oxidation and isotype reactions. These findings are significant for our understanding of the metabolism of ZSF. The proposed metabolic pathways of bioactive components might be crucial for further studies of the mechanisms of action and pharmacokinetic evaluations of ZSF.

Zi Shen Wan Fang Attenuates Neuroinflammation and Cognitive Function Via Remodeling the Gut Microbiota in Diabetes-Induced Cognitive Impairment Mice

“…These results suggest that the reduction of peripheral circulation and neuroinflammation is the mechanism of ZSWF to improve DCI. However, studies showed that the bioavailability of main chemical components of Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex is low (Singh and Chaudhuri, 2018;Baldim et al, 2020), and our previous in vivo chemical analysis of ZSWF also showed that most components in Cistanches Herba were concentrated in feces (Zheng et al, 2018), suggesting that regulation of gastrointestinal function or intestinal flora may play a crucial role in the reversal of DCI inflammation by ZSWF.…”

“…However, this study still demonstrates some limitations. For example, although our previous studies identified the main chemical components of ZSWF extract in vivo and in vitro, including the main archetypes and metabolic components in feces (Zheng et al, 2018), which components are involved in maintaining intestinal microbiota homeostasis and their mechanisms remain unclear. Moreover, since no drug with a definite therapeutic effect for DCI exists in clinical practice at present, this study did not select a suitable positive control.…”

“…The drug materials of Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex were purchased from Hebei anguo Chinese Herbal Medicine Co., Ltd., and Cistanches Herba was purchased from Inner Mongolia Mandera Biological Technology Co., Ltd. These herbs were identified by professor Tianxiang Li (TianJin University of Traditional Chinese Medicine) and were extracted according to our previous method (Zheng et al, 2018). In simple terms, Anemarrhenae Rhizoma and Phellodendri Chinensis Cortex were mixed at the ratio of 1: 1 by weight, then extracted by reflux for 3 times with 80% ethanol of 8 times (2 h each time), and filtrate was collected.…”

“…Next, the volume of liquid required for ingastric administration of mice during the treatment cycle was calculated, and all filtrate was mixed and concentrated to 0.94 g/ml ZSWF crude extract by rotary evaporator, which was separated and stored in a refrigerator at −80 °C for use. Moreover, our previous study systematically investigated the main chemical components of ZSWF extract (Zheng et al, 2018), including in vitro components (e.g., Neomangiferin, Berberine, TimosaponinBⅡ, and Cistansinenside A), absorbed components (e.g., Mangiferin, Timosaponin C, and 3/5-O-Feruloylquinic acid), as well as prototypical (e.g., Phellodenrine, Tetrahydropalmatine, and Oxoberberine) and metabolic components [e.g., 3,4dihydroxybenzenepropionic acid and (20R, 25S)-timosaponin AI] in feces.…”