OPTIMIZATION OF LC-MS/MS METHOD FOR THE SIMULTANEOUS DETERMINATION OF METFORMIN AND ROSIGLITAZONE IN HUMAN PLASMA WITH BOX-BEHNKEN DESIGN

RUBINA KAUSER; SUNIL KUMAR CHAITANYA PADAVALA; VENKATESAN PALANIVEL

doi:10.22159/ijap.2024v16i6.51936

Authors

RUBINA KAUSER Department of Pharmacy, Annamalai University, Annamalai Nagar–608002, Chidambaram, Tamil Nadu, India https://orcid.org/0009-0009-3098-3312
SUNIL KUMAR CHAITANYA PADAVALA Department of Pharmaceutical Analysis and Quality Assurance, St. Pauls College of Pharmacy, Hyderabad–501510, Telangana, India
VENKATESAN PALANIVEL Department of Pharmacy, Annamalai University, Annamalai Nagar–608002, Chidambaram, Tamil Nadu, India https://orcid.org/0000-0002-0601-7861

DOI:

https://doi.org/10.22159/ijap.2024v16i6.51936

Keywords:

Metformin, Rosiglitazone, Box-behnken design, Biological matrices, Optimization, LC-MS/MS

Abstract

Objective: The objective of this study was to develop a robust Liquid Chromatography – Tandem Mass Spectrometry (LC-MS/MS) methodology for the precise quantification of metformin and rosiglitazone in human plasma.

Methods: A Design of Experiments (DOE) framework was utilized, specifically employing a Box-Behnken experimental design, to optimize critical parameters such as Capillary voltage, Cone voltage, Desolvation temperature, and Collision energy. Sample preparation involved protein precipitation using acetonitrile, simplifying the procedure. Chromatography was performed with a mobile phase of 0.1% formic acid and acetonitrile (60:40 V/V) to enhance sensitivity and reproducibility. Quantification was achieved through Multiple Reaction Monitoring (MRM) of the transition’s m/z 130.1 → m/z 60.1 for metformin, m/z 358.2 → m/z 134.9 for rosiglitazone, and m/z 206.3 → m/z 59.9 for phenformin. The methodology was validated according to regulatory guidelines.

Results: The developed methodology demonstrated selectivity, linearity, accuracy, precision, recovery, and stability. The calibration curve showed linearity over the concentration range of 5 ng/ml to 1000 ng/ml for metformin and 1.5 ng/ml to 300 ng/ml for rosiglitazone. Accuracy and precision were within acceptable limits across calibration and quality control standards. Assessments of extraction recovery and matrix effects confirmed the robustness of the extraction procedure, with negligible interference from plasma components. Stability studies indicated that the method maintained acceptable limits for metformin and rosiglitazone concentrations under various storage and handling conditions.

Conclusion: The validated Liquid Chromatography – Tandem Mass Spectrometry (LC-MS/MS) methodology provides a reliable and accurate platform for the quantification of metformin and rosiglitazone in human plasma. This method shows potential applications in pharmacokinetic studies and clinical research, ensuring consistent performance in routine analysis.

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