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Current Chromatography

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ISSN (Print): 2213-2406
ISSN (Online): 2213-2414

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Research Article

Development and Validation of a Simple Micromethod for the Determination of Metformin in Plasma by HPLC-UV for Application in Pharmacokinetic Studies

Author(s): Miriam N. Morales-Barragán, María del Carmen Romero-Méndez, Rosa del Carmen Milán-Segovia, Jaime G. Rodríguez-Rivera, María A. Aguilar-Torres and Silvia Romano-Moreno*

Volume 8, Issue 1, 2021

Published on: 11 August, 2021

Article ID: e110821195554 Pages: 8

DOI: 10.2174/2213240608666210811123051

Price: $65

Abstract

Background: Metformin is the first-line drug to enhance glycemic control of type 2 DIABETES Mellitus (DM2) patients. Some reported methods to determine plasma metformin by HPLC-UV are not sensitive enough. Other methods require long extraction processes.

Objective: The objective of this study was to develop and validate a simple and rapid analytical method to determine plasma metformin by HPLC-UV for application in a population pharmacokinetic study.

Methods: Analyte was extracted from plasma by a simple protein precipitation technique using trichloroacetic acid (15%, w/v) as the precipitating agent. Plasma samples were analyzed using a C18 column (3.0 x 150 mm, 3.5 μm) under isocratic elution with 30 mM sodium hexansulfonate (pH 5) and acetonitrile (95: 5, v,v).

Results: The limit of quantification (LOQ) was 0.1 μg mL-1 and the calibration curve was linear up to 4 μg mL-1 with a correlation coefficient >0.99. The mean recovery for metformin using this extraction procedure was 84.4 - 86.6%. The intra- and inter-day coefficients of variation and percent error values of the assayed method were <20% and <15% for LOQ and QCs, respectively. Metformin was stable in plasma samples by subjecting it to three freeze-thaw cycles and storing it up to 60 days at -80°C. This method was applied to determine plasma metformin concentrations in patients with type 2 diabetes mellitus treated with this drug.

Conclusion: The HPLC-UV method developed is selective, accurate and precise for the quantification of metformin in plasma samples, since sample processing is fast and simple, in addition to being applicable in pharmacokinetic studies.

Keywords: Metformin, HPLC, plasma, C18 column, pharmacokinetics, diabetes.

Graphical Abstract

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