International Journal of Drug Delivery Technology
Volume 16, Issue 4, 2026

Design and Optimization of LC-MS Bioanalytical Method for Efonidipine and Chlorthalidone Using Quality by Design Strategy

Rajasekaran C1, Sudha T2*

1 Research Scholar, Department of Chemistry and Analysis, Vels Institute of Science, Technology and Advance Studies (VISTAS), Pallavaram, Tamilnadu, India. ORCID: https://orcid.org/0000-0002-4593-1541

2* Associate Professor, Department of Pharmaceutical Chemistry and Analysis, School of Pharmaceutical Sciences, Vels Institute of Science, Technology and Advanced Studies (VISTAS), Chennai, Tamilnadu, India (Corresponding Author). ORCID: https://orcid.org/0000-0001-8821-3999

Received Date: 10th October 2025; Revised Date: 25th January 2026; Accepted Date: 5th April 2026
📊 Graphical Abstract — (Illustrative summary of the LC-MS method development and optimization using Quality by Design strategy)

ABSTRACT

Objective

The Quality by Design (QbD) technique was used in this work to design and validate a novel LC-MS method for the simultaneous measurement of Efonidipine Hydrochloride, Ethanolate, and Chlorthalidone in artificially generated human plasma. To guarantee the method achieved the right analytical performance, the parameters of flow rate, heat block temperature, and mobile phase composition in methanol were optimised in an analytical science, systematic approach employing a central composite design. Resolution and retention time were two of the analytical method's most important characteristics.

Methods

The chromatographic separation was achieved using a Shimadzu capcell pak C18 (150×4.6mm, 3µm), and a mobile phase consisting of Phosphate buffer (pH 3.0) (40:35:25% v/v/v): methanol and acetonitrile. The mobile phase was allowed to flow at a rate of 1.0 mL/min. A mass spectrometer coupled with an electrospray ionization (ESI) source operating in the positive ion was used for detection. Data were obtained in the multi-reaction monitoring (MRM) acquisition mode. The best recovery and maximum matrix cleanliness were obtained from the liquid-liquid extraction sample preparation.

Results

The retention time for Efonidipine and Chlorthalidone were 6.38 and 8.60 min respectively. Sample extraction was performed using liquid-liquid extraction (LLE), and this technique produced very pure extracts with good recovery rates. A linear calibration curve was found in the range of 18.75-65.61 µg/mL for Efonidipine and 60-210 µg/mL for Chlorthalidone with a correlation coefficient r² > 0.99. The validation process confirmed the LC-MS analytical method's selectivity, sensitivity, linearity, accuracy, and precision for both Chlorthalidone and Efonidipine. Bench-top stability, freeze-thaw stability, short-term stability, long-term stability, and autosampler stability data showed good recovery and proved the method's stability.

Conclusion

Overall, it was shown that the optimised LC-MS analytical approach was dependable and might be helpful for routine work in pharmacokinetic, bioequivalence, and quality control investigations for medications like Chlorthalidone and Efonidipine.

Keywords: LC-MS, QbD, Efonidipine Hydrochloride Ethanolate, Chlorthalidone, Artificial Human Plasma.

How to cite this article: Rajasekaran C, Sudha T. Design and Optimization of LC-MS Bioanalytical Method for Efonidipine and Chlorthalidone Using Quality by Design Strategy. Int J Drug Deliv Technol. 2026;16(4): 284-294. DOI: 10.25258/ijddt.16.4.30

Source of support: Nil.

Conflict of interest: None