Study on the metabolites of betulinic acid in vivo and in vitro by ultra high performance liquid chromatography with time‐of‐flight mass spectrometry

Abstract

Betulinic acid is a triterpenoid organic acid with remarkable antitumor properties and is naturally present in many fruits, condiments and traditional Chinese medicines. Currently, a strategy was developed for the identification of metabolites following the in vivo and in vitro biotransformation of Betulinic acid with rat intestinal bacteria utilizing ultra high performance liquid chromatography with time‐of‐flight mass spectrometry with polymeric solid‐phase extraction. As a result, 46 metabolites were structurally characterized. The results demonstrated that Betulinic acid is universally metabolized in vivo and in vitro, and Betulinic acid could undergo general metabolic reactions, including oxidation, methylation, desaturation, loss of O and loss of CH2. Additionally, the main metabolic pathways in vivo and in vitro were determined by calculating the relative content of each metabolite. This is the first study of Betulinic acid metabolism in vivo, whose results provide novel and useful data for better understanding of the safety and efficacy of Betulinic acid.

Structural Investigation of Betulinic Acid Plasma Metabolites by Tandem Mass Spectrometry

“…The screening mass spectra of analyzed samples indicated the presence of the precursor ion identified as [M-H] − at m / z 469.37 ( M5 ),which has a Mw 14 Da more than BA, and was assigned according to the mass calculations to the methylated metabolite of BA, which is consistent with previous findings that reported methylated metabolites for echinocystic acid, a triterpene [ 39 ], as well as for BA [ 22 , 23 ]. Although methylation is a minor conjugation reaction for xenobiotic transformation, being more common for endogenous neurotransmitters, it still remains a metabolic pathway for a large number of medicinal drugs and phytocompounds [ 38 ].…”

“…The available data regarding the in vivo metabolism of BA are even more limited, hence only a couple of studies have been published. Recently, Zhang et al [ 22 ] reported on some phase I metabolites identified in the feces of Sprague-Dawley rats after oral administration of BA-carboxyl methyl cellulose solution. Another study conducted by Li et al [ 23 ] reported on other phase I and phase II metabolites of BA mainly identified in the feces of Sprague-Dawley rats after oral administration of BA, spinosin, isovitexin and jujuboside together with Ziziphi Spinosae semen extract.…”

A Systematic Study of the Metabolites of Dietary Acacetin in Vivo and in Vitro Based on UHPLC-Q-TOF-MS/MS Analysis

“…The relative quantitative estimation of metabolites was carried out on the basis of the peak areas of XICs. The area normalization method was utilized to estimate relative quantification of metabolites . Then, the peak area ratio ( W %) of each metabolite in a specific matrix was expressed as follows: W % = the peak area of each metabolite/the peak area of parent drug and all metabolites × 100% in the matrix.…”

“…The area normalization method was utilized to estimate relative quantification of metabolites. 33 Then, the peak area ratio (W%) of each metabolite in a specific matrix was expressed as follows: W% = the peak area of each metabolite/ the peak area of parent drug and all metabolites × 100% in the matrix. The peak areas of acacetin and metabolites that were detected in blood, bile, urine, feces, and microsomes were calculated and are listed in Supplementary Table 1.…”

Metabolites study on 5‐O‐methylvisammioside in vivo and in vitro by ultra high performance liquid chromatography coupled to quadrupole time‐of‐flight mass spectrometry

“…In this study, it was the first time to develop a rapid and reliable method to analyze and identify the metabolites of 5-O-methylvisammioside in vivo and in vitro by combining UHPLC-Q-TOF-MS/MS with multiple data processing technologies. In addition, area normalization method was applied to evaluate the relative contents of metabolites in each biological sample for exploring the major metabolic pathway [17]. As a result, based on accurate masses, chromatographic behaviors and MS/MS fragment ions, 26 metabolites in vivo and seven metabolites in vitro were detected, and the metabolic pathways were summarized and compared.…”