HPLC MS/MS method development for the quantitative determination of verapamil hydrochloride from Caco-2 cell monolayers release_37s4nft2nzeyvni7b2oreu5utu

by Liliya Logoyda, Maksym Herasymiuk, Dariya Popovych, Svitlana Pidruchna, Vitaliy Hlushok, Nazar Herasymiuk, Nadiya Zarivna

Published in Farmatsiia by Pensoft Publishers.

2020   Volume 67, p63-69

Abstract

<jats:bold>Aim.</jats:bold> An understanding of the role that transporters, in particular P-glycoprotein (P-gp), can play in the absorption, distribution, metabolism and excretion (ADME) of candidate drugs, and an assessment of how these processes might impact on toxicity and the potential for drug-drug interactions in the clinic, is required to support drug development and registration. It is therefore necessary to validate preclinical assays for the in vitro evaluation of candidate drugs as substrates or inhibitors of human P-gp. 2. A simple, rapid HPLC MS/MS method was developed for determination of verapamil hydrochloride from confluent Caco-2 monolayers and from aqueous solution. <jats:bold>Materials and methods.</jats:bold> Chromatography was achieved on Discovery C18, 50 × 2.1 mm, 5 μm column. Samples were chromatographed in a gradient mode (eluent A (acetonitrile – water – formic acid, 5 : 95 : 0.1 v/v), eluent B (acetonitrile – formic acid, 100 : 0.1 v/v)). The initial content of the eluent B is 0%, which increases linearly by 1.0 min to 100% and to 1.01 min returns to the initial 0%. The mobile phase was delivered at a flow rate of 0.4 mL/min into the mass spectrometer ESI chamber. The sample volume was 5 μl. <jats:bold>Results.</jats:bold> Under these conditions, verapamil hydrochloride was eluted at 1.08 min. A linear response function was established at 1 – 100 ng/mL. The regression equation for the analysis was Y = 0.0162x + 0.00391 with coefficient of correction (R<jats:sup>2</jats:sup>) = 0.9992. According to the Caco-2 test results, verapamil showed low permeability. It should be noted that the recovery value for verapamil hydrochloride is 102.69%. The within-run coefficients of variation ranged between 0.336% and 0.617% for verapamil. The within-run percentages of nominal concentrations ranged between 98.82% and 100.62% for verapamil. The between-run coefficients of variation ranged between 0.334% and 0.612% for verapamil. The between-run percentages of nominal concentrations ranged between 98.97% and 101.76% for verapamil. The assay values on both the occasions (intra- and inter-day) were found to be within the accepted limits. <jats:bold>Conclusion.</jats:bold> From results of analysis, it can be concluded that developed method is simple and rapid for determination of verapamil hydrochloride from confluent Caco-2 monolayers and from aqueous solution. Acquired results demonstrate that proposed strategy can be effortlessly and advantageously applied for examination of verapamil hydrochloride from Caco-2 cell monolayers.
In application/xml+jats format

Archived Files and Locations

application/pdf  376.4 kB
file_czue6rrnojgxzmwx5jbsp67t4a
pharmacia.pensoft.net (publisher)
web.archive.org (webarchive)
Read Archived PDF
Preserved and Accessible
Type  article-journal
Stage   published
Date   2020-07-31
Container Metadata
Open Access Publication
In DOAJ
In Keepers Registry
ISSN-L:  0428-0296
Work Entity
access all versions, variants, and formats of this works (eg, pre-prints)
Catalog Record
Revision: 9deb2339-85e4-4a17-aa3c-237c84b8c739
API URL: JSON