High-Resolution Mass Spectrometry in Metabolite Identification

Adhikari

BioMed Research International

et al. 2021

Acacia catechu (L.f.) Willd is a profoundly used traditional medicinal plant in Asia. Previous studies conducted in this plant are more confined to extract level. Even though bioassay-based studies indicated the true therapeutic potential of this plant, compound annotation was not performed extensively. This research is aimed at assessing the bioactivity of different solvent extracts of the plant followed by annotation of its phytoconstituents. Liquid chromatography equipped with high resolution mass spectrometry (LC-HRMS) is deployed for the identification of secondary metabolites in various crude extracts. On activity level, its ethanolic extract showed the highest inhibition towards α-amylase and α-glucosidase with an IC50 of 67.8 ± 1 μg/mL and 10.3 ± 0.1 μg/mL respectively, inspected through the substrate-based method. On the other hand, the plant extract showed an antioxidant activity of 23.76 ± 1.57 μg/mL, measured through radical scavenging activity. Similarly, ethyl acetate and aqueous extracts of A. catechu showed significant inhibition against Staphylococcus aureus with a zone of inhibition (ZoI) of 13 and 14 mm, respectively. With the LC-HRMS-based dereplication strategy, we have identified 28 secondary metabolites belonging to flavonoid and phenolic categories. Identification of these metabolites from A. catechu and its biological implication also support the community-based usage of this plant and its medicinal value.

supporting

confidence: 67%

Section: Introductionmentioning

confidence: 99%

LC-HRMS Profiling and Antidiabetic, Antioxidant, and Antibacterial Activities of Acacia catechu (L.f.) Willd

Adhikari

BioMed Research International

et al. 2021

“…By comparison, with databases or by the application of rulesets, such as isotope pattern analysis or data filtering by metabolism-associated nominal mass shifts [ 16 ], the data from HR-MS spectra can be filtered, such that the chromatographic features can be associated with sum formulae. If sum formula data are put into the context of compound/metabolite classes (e.g., the metabolites of a parent compound or lipids of a certain lipid class), the tentative assignment of a chromatographic peak as a molecule of interest is feasible [ 17 , 18 ].…”

Section: Introductionmentioning

confidence: 99%

Combining HPLC-DAD-QTOF-MS and HPLC-SPE-NMR to Monitor In Vitro Vitetrifolin D Phase I and II Metabolism

et al. 2021

By combining HPLC-DAD-QTOF-MS and HPLC-SPE-NMR, the in vitro metabolism of vitetrifolin D, a pharmacologically active key molecule from Vitex agnus-castus in liver cell fractions, was investigated. Twenty-seven phase I and phase II metabolites were tentatively identified from the culture broth by HPLC-DAD-QTOF-MS. The subsequent HPLC-SPE-NMR analysis allowed for the unequivocal structural characterization of nine phase I metabolites. Since the preparative isolation of the metabolites was avoided, the substance input was much lower than in conventional strategies. The study did prove that the use of hyphenated instrumental analysis methodologies allows for the successful performance of in vitro metabolism studies, even if the availability of substances is very limited.

Drug Testing and Analysis

“…The main objective of this study was to perform detailed metabolite profiling of abemaciclib and to detect and characterize its metabolites generated in in vitro (RLM, HLM, RS9) and in vivo biological matrices (rat plasma, urine, and feces) using high‐resolution mass spectrometry (HRMS) coupled with liquid chromatography (LC). LC‐HRMS is the first preference of researchers for the identification and characterization of metabolites . Additionally, an in silico study was performed to predict the vulnerable metabolic site of abemaciclib.…”

Section: Introductionmentioning

confidence: 99%

“…LC-HRMS is the first preference of researchers for the identification and characterization of metabolites. 10,11 Additionally, an in silico study was performed to predict the vulnerable metabolic site of abemaciclib.…”

mentioning

confidence: 99%

Update on metabolism of abemaciclib: In silico, in vitro, and in vivo metabolite identification and characterization using high resolution mass spectrometry

Thakkar

Kate

2020

Abemaciclib was approved by the US Food and Drug Administration in 2015 as an advanced treatment for metastatic breast cancer. Identification and characterization of limited numbers of abemaciclib metabolites have been reported in the literature. Therefore, the current study focused on the investigation of the in vitro and in vivo metabolic fate of abemaciclib using high resolution mass spectrometry. Initially, a vulnerable site of metabolism was predicted by the Xenosite web predictor tool. Later, in vitro metabolites were identified from pooled rat liver microsomes, rat S9 fractions, and human liver microsomes. Finally, in vivo metabolites have been detected in plasma, urine, and feces matrix of male Sprague–Dawley rats. A total of 12 putative metabolites (11 phase I and 1 phase II) of abemaciclib and their metabolic pathways were proposed by considering accurate mass, mass fragmentation pattern, nitrogen rule, and ring double bonds of the detected metabolites. Abemaciclib was metabolized via hydroxylation, N‐oxidation, N‐dealkylation, oxidative deamination followed by reduction and sulfate conjugation. In the human liver microsomes, maximum numbers of metabolites (11 metabolites) were observed, from which M7, M8, M9, and M11 were human specific.