Identification and characterization of fluvastatin metabolites in rats by UHPLC/Q‐TOF/MS/MS and in silico toxicological screening of the metabolites

Abstract

The present study reports the in vivo and in vitro identification and characterization of metabolites of fluvastatin, the 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase inhibitor, using liquid chromatography-mass spectrometry (LC-MS). In vitro studies were conducted by incubating the drug with human liver microsomes and rat liver microsomes. In vivo studies were carried out by administration of the drug in the form of suspension to the Sprague-Dawley rats followed by collection of urine, faeces and blood at different time points up to 24 h. Further, samples were prepared by optimized sample preparation method, which includes freeze liquid extraction, protein precipitation and solid phase extraction. The extracted and concentrated samples were analysed using ultrahigh-performance liquid chromatography-quadruple time-of-flight tandem mass spectrometry. A total of 15 metabolites were observed in urine, which includes hydroxyl, sulphated, desisopropyl, dehydrogenated, dehydroxylated and glucuronide metabolites. A few of the metabolites were also present in faeces and plasma samples. In in vitro studies, a few metabolites were observed that were also present in in vivo samples. All the metabolites were characterized using ultrahigh-performance liquid chromatography-quadruple time-of-flight tandem mass spectrometry in combination with accurate mass measurement. Finally, in silico toxicity studies indicated that some of the metabolites show or possess carcinogenicity and skin sensitization. Several metabolites that were identified in rats are proposed to have toxicological significance on the basis of in silico evaluation. However, these metabolites are of no human relevance. Copyright © 2017 John Wiley & Sons, Ltd.

In vivo and in vitro metabolite profiling of nirmatrelvir using LC–Q‐ToF–MS/MS along with the in silico approaches for prediction of metabolites and their toxicity

“…The accurate masses of these two metabolites were m/z 420.2623 and m/z 420.2623 as shown in the MS/MS spectra in Figure 1b,c. The elemental composition of C 21 H 34 N 5 O 4 + indicates that there is a loss of two carbons and three fluorine atoms, suggesting that these metabolites were formed by the hydrolysis of an amide bond followed by hydroxylation (Chavan et al, 2017). The observed metabolic change was justified by the fragmentation pattern, which is as follows (Figure 5): as per the MS/MS spectrum, M1 initially dissociated to two product ions, namely, m/z 402 on the loss of H 2 O and m/z 307 on the loss of C 6 H 11 NO.…”

A novel microextraction technique aided by air agitation using a natural hydrophobic deep eutectic solvent for the extraction of fluvastatin and empagliflozin from plasma samples: application to pharmacokinetic and drug–drug interaction study

“…The combination of uorescence detection with extraction using hydrophobic deep eutectic solvents (DES) for the analysis of studied drugs offers several advantages over traditional techniques such as high-performance liquid chromatography (HPLC), [15][16][17][18][19][20] electrochemical techniques, [21][22][23] and spectroscopic methods. [24][25][26][27][28][29] Firstly, uorescence detection provides high sensitivity and selectivity, allowing for the detection and quantication of drugs at low concentrations.…”

Comprehensive metabolite identification study of arterolane using hydrophilic interaction liquid chromatography with quadrupole‐time‐of‐flight mass spectrometry

“…In vitro studies involving microsomal enzymes and in vivo animal studies are usually applied to understand drug metabolism 3 . In recent times, the site of metabolism has been predicted through in silico studies and probable structures of metabolites are being identified quickly by hyphenated techniques like liquid chromatography/high‐resolution mass spectrometry (LC/HRMS) 4–6 …”

“…3 In recent times, the site of metabolism has been predicted through in silico studies and probable structures of metabolites are being identified quickly by hyphenated techniques like liquid chromatography/high-resolution mass spectrometry (LC/HRMS). [4][5][6] Arterolane (ART) is reported as one of the fast-acting schizonticides at the erythrocytic stages of the malaria parasite which is being approved in India to treat plasmodium-associated malaria.…”