ICRF-Assisted Box-Behnken Design and Optimization for Rapid UPLC-PDA Determination of Dorzolamide Hydrochloride and Timolol Maleate in an Ophthalmic Preparation.

Authors

Erdal Dinç, Eda Büker

Published in

Journal of AOAC International. Jun 25, 2026. Epub Jun 25, 2026.

Abstract

Simultaneous optimization of chromatographic resolution and analysis time constitutes a significant analytical challenge in multicomponent pharmaceutical analysis, as resolution-driven optimization strategies may improve peak separation without providing explicit control over retention time, often resulting in unnecessarily prolonged runtimes. To address this limitation, an optimization strategy based on a Box-Behnken experimental design was implemented in conjunction with the Improved Chromatographic Response Function (ICRF), which integrates separation quality and analysis time within a single mathematical objective function.
This strategy describes the development of a rapid, chemometrically optimized UPLC-PDA method for the simultaneous determination of dorzolamide hydrochloride (DH) and timolol maleate (TI) in a commercial ophthalmic preparation.
A Box-Behnken experimental design and optimization approach was employed in combination with an Improved Chromatographic Response Function (ICRF), which integrates resolution, peak overlap, peak width, and runtime into a single composite objective function. This strategy enabled short runtime (or short retention time of analytes in a chromatogram) while preserving adequate peak separation.
Under the optimized conditions, complete chromatographic separation was achieved within 3 min using a BEH C18 column and a mobile phase consisting of acetonitrile and 4 × 10-4 M CCl3COOH (60:40, v/v) at a flow rate of 0.32 mL/min with detection at 275 nm. The method demonstrated excellent linearity over the range of 5.0-40.0 µg/mL (r > 0.999), with limits of detection of 0.51 µg/mL for DH and 0.61 µg/mL for TI. Mean recoveries were 99.9% and 99.5% for DH and TI, respectively, with satisfactory precision and robustness.
The proposed ICRF-assisted optimization approach provided high-resolution separation within minimal runtime and was successfully applied to the routine quality-control analysis of a commercial ophthalmic formulation.
The study demonstrates the effectiveness of composite response-based chemometric optimization in enhancing analytical efficiency in pharmaceutical drug analysis.

PMID:
42348236
Bibliographic data and abstract were imported from PubMed on 25 Jun 2026.

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