In vivo metabolite identification of acotiamide in rats using ultra‐performance liquid chromatography–quadrupole/time‐of‐flight mass spectrometry

Abstract

Acotiamide hydrochloride (ACT) is a drug used for the treatment of functional dyspepsia. Understanding which metabolites are likely to be formed in vivo is essential for interpreting pharmacology, pharmacokinetic and toxicology data. The metabolism of ACT has been investigated using a specific and sensitive liquid chromatography positive ion electrospray ionization high-resolution tandem mass spectrometry method. In vivo samples including rat plasma, urine and feces were collected separately after dosing healthy Sprague-Dawley rats at a dose of 20 mg kg ACT at different time points up to 24 h. The metabolites were enriched by optimized sample preparation involving protein precipitation using acetonitrile followed by solid-phase extraction. The mass defect filter technique was used for better detection of both predicted and unexpected drug metabolites with the majority of interference ions removed. The structural elucidation of the metabolites was performed by comparing their [M + H] ions and their product ions with those of the parent drug. As a result, a total of seven hitherto unknown metabolites were characterized from the biosamples. The only phase I metabolite detected was N-despropyl acotiamide, whereas six phase II glucuronide conjugate metabolites were identified.

Development and Validation of Stability Indicating Method Rp-HPLC Method of Acotiamide

“…Stress testing helps in identification of degradation products and degradation behaviour of drug [5,6]. The literature reveals there were few analytical methods available for determination of acotiamide by HPLC [7], LC-MS-MS [8], UPLC-Q-TOF-MS [9,10], stability indicating the method by UPLC-Q-TOF-MS (gradient mode) [11], this reported method based on UPLC method.…”

Stability‐indicating assay method for acotiamide: Separation, identification and characterization of its hydroxylated and hydrolytic degradation products along with a process‐related impurity by ultra‐high‐performance liquid chromatography/electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry

“…As the drug was only approved recently, no reports were found in the literature on SIAM for ACM. Recently, we reported pharmacokinetic studies and characterization of metabolites of ACT in rat biological samples . In the present study, we have developed a SIAM for ACM and identified the degradation products of ACM by using ultra‐high‐performance liquid chromatography/electrospray ionization quadrupole time‐of‐flight tandem mass spectrometry (UHPLC/ESI‐QTOF‐MS/MS).…”

Identification, Separation and Characterization of Potential Degradation Products in Acotiamide Drug Substance

“…The presence of impurities can have a significant impact on the product quality, safety and efficacy, hence the percentage level of impurities need to control in the drug substance as well as a drug product 5,6 . The literature review reveals that there were few analytical methods available for determination of acotiamide by HPLC 7 , LC-MS-MS 8 , UPLC-Q-TOF-MS 9,10 , identification of degradation products of acotiamide by UPLC/ESI-quadrupole TOF-tandem MS 11 .…”