Screening of Intestinal Bacterial Metabolites of Platycodin D Using Ultra-Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry

Abstract

Platycodin D (PD), a bioactive triterpenoid saponin isolated from Platycodi Radix (PR), possesses a vast range of biological activities. Although the pharmacological activities and pharmacokinetics of PD have been well demonstrated, information regarding the intestinal metabolisms of PD is very limited. In this study, human and rat fecal microflora were prepared and anaerobically incubated with PD at 37[Formula: see text]C for 48[Formula: see text]h, respectively. A highly sensitive and specific ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was developed for the analysis of PD and related metabolites in the reaction samples. A Liquid-liquid extraction method was used for sample pretreatment and the chromatographic separation was performed on a 1.7 [Formula: see text]m particle size Syncronis C[Formula: see text] column using gradient elution system. Finally, a total of seven metabolites were detected and tentatively identified, such as the demethylation metabolite (M1), deoxidation metabolites (M3, M7) and hydrolysis at the C-28 oligosaccharide metabolites (M5, M6), which were first discovered in this experiment. The results indicate that hydrolysis, demethylation, dehydroxylation, and acetylation were the major metabolic pathways of PDin vitro. Additionally, four bacterial strains from human feces including Enterococcus sp.41, Bacillus sp.46, Escherichia sp.49 A and Escherichia sp.64 were detected and further identified with 16S rRNA gene sequencing due to their relatively strong metabolic capacity toward PD. The present study provides important information about the metabolism of PD, which will help elucidate the impact of intestinal bacteria on this active component.

Fecal metabolomic dataset of American ginseng-treated DSS mice: Correlation between ginseng enteric inflammation inhibition and its biological signatures

“…Stool samples were extracted using water and methanol with homogenization [12] , [14] . Before analysis, a two-step derivation process was conducted using methoxyamine and BSTFA containing 1% TMCS.…”

Rapid Screening of Chemical Constituents in Rhizoma Anemarrhenae by UPLC-Q-TOF/MS Combined with Data Postprocessing Techniques

“…At present, the analytical methods of TCM ingredients mainly include thin layer chromatography (TLC), gas chromatography (GC), high-performance liquid chromatography (HPLC), and ultraperformance liquid chromatography/time-of-flight mass spectrometry (UPLC-Q-TOF/MS), among others. As a comprehensive analysis technique, UPLC-Q-TOF/MS has been increasingly applied for the qualitative and quantitative analysis of components in TCMs and compound preparations [6–11]. However, this method displays several disadvantages, including complicated and time-consuming data processing.…”

Metabolite identification and pharmacokinetic study of platycodi radix (Jiegeng) in vivo

“…The metabolites of platycosides in vivo all contained glucoses at C-3, but in vitro the glucoses could be easily hydrolyzed by enzyme that produced by intestinal bacteria; the secondary platycosides without saccharide chain at C-28 were the main absorbed compounds in vivo, but the saccharide chain at C-28 cloud not be enzymatic hydrolyzed in vitro and the main metabolites of platycosides in vitro were the compounds with the acetylated saccharide at C-28. 17,24,35 So the hydrolysis of saccharide chain at C-28 by gastric acid in the stomach was considered to be the main reason of the degradations of primary platycosides and the differences between the metabolites in vitro and in vivo. By comparing the relative contents obtained from peak areas in EIC, and considering the absorption and degradation of saponins, the two secondary saponins with a glucosyl linked at C-3, including GP and GPA, were used as the representative compounds in pharmacokinetic analysis.…”