Determination of metformin in rat plasma by HILIC‐MS/MS combined with Tecan automation and direct injection
- 27 December 2011
- journal article
- Published by Wiley in Biomedical Chromatography
- Vol. 26 (10), 1163-1169
- https://doi.org/10.1002/bmc.2673
Abstract
Metformin is a well‐known oral antihyperglycemic drug used in treatment of type II diabetes. Analysis of metformin in biological fluids is a challenge owing to its high polarity and small molecular size, which lead to poor retention of metformin on reversed‐phase liquid chromatographic columns. A high‐throughput method was developed and validated for the determination of metformin in rat plasma in support of preclinical toxicology studies, using hydrophilic interaction liquid chromatography tandem mass spectrometry (HILIC‐MS/MS) and Tecan automated sample preparation. Extracted samples were directly injected onto the unbounded silica column with an aqueous–organic mobile phase. This HILIC‐MS/MS method was validated for accuracy, precision, sensitivity, stability, matrix effect, recovery and calibration range. Acceptable intra‐run and inter‐run assay precision (coefficient of variation ≤ 3.9%) and accuracy (99.0–101.8%) were achieved over a linear range of 50–50,000 ng/mL. Metformin is stable in rat plasma for at least 6 h at room temperature, 147 days at −70°C and through three freeze (−70°C) and thaw cycles. Metformin is also stable in rat whole blood for at least 2 h at room temperature and in an ice–water bath. The validated method was successfully used in support of several preclinical studies where metformin is dosed together with an investigational drug substance. The ruggedness of the validated method was demonstrated by the incurred sample reproducibility test. Copyright © 2011 John Wiley & Sons, Ltd.Keywords
This publication has 19 references indexed in Scilit:
- LC–MS/MS method using unbonded silica column and aqueous/methanol mobile phase for the simultaneous quantification of a drug candidate and co-administered metformin in rat plasmaJournal of Chromatography B, 2010
- Recent advances in application of hydrophilic interaction chromatography for quantitative bioanalysisJournal of Separation Science, 2010
- LC–MS–MS Development and Validation for Simultaneous Quantitation of Metformin, Glimepiride and Pioglitazone in Human Plasma and Its Application to a Bioequivalence StudyChromatographia, 2009
- Standards of Medical Care in Diabetes—2009Diabetes Care, 2009
- Liquid chromatography tandem mass spectrometry method for simultaneous determination of antidiabetic drugs metformin and glyburide in human plasmaJournal of Pharmaceutical and Biomedical Analysis, 2007
- Development and validation for high selective quantitative determination of metformin in human plasma by cation exchanging with normal-phase LC/MS/MSJournal of Pharmaceutical and Biomedical Analysis, 2005
- Direct injection of solid‐phase extraction eluents onto silica columns for the analysis of polar compounds isoniazid and cetirizine in plasma using hydrophilic interaction chromatography with tandem mass spectrometryRapid Communications in Mass Spectrometry, 2004
- Rapid determination of metformin in human plasma by liquid chromatography−tandem mass spectrometry methodJournal of Chromatography B, 2004
- Metformin: its botanical backgroundPractical Diabetes International, 2004
- Importance of injection solution composition for LC–MS–MS methodsJournal of Pharmaceutical and Biomedical Analysis, 2001