ICRF-assisted Box–Behnken design and optimization for rapid UPLC–PDA determination of dorzolamide hydrochloride and timolol maleate in an ophthalmic preparation
Abstract
Background: 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.
Objective: This strategy describes the development of a rapid, chemometrically optimized Ultra Performance Liquid Chromatography coupled with a Photodiode Array Detector (UPLC–PDA) method for the simultaneous determination of dorzolamide hydrochloride (DH) and timolol maleate (TI) in a commercial ophthalmic preparation.
Methods: A Box–Behnken experimental design and optimization approach was employed in combination with an 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.
Results: Under the optimized conditions, complete chromatographic separation was achieved within 3 min using a BEH C18 column and a mobile phase consisting of acetonitrile–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 LODs 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.
Conclusion: The proposed ICRF-assisted optimization approach provided high-resolution separation within minimal runtime and was successfully applied to the routine QC analysis of a commercial ophthalmic formulation.
Highlights: The study demonstrates the effectiveness of composite response-based chemometric optimization in enhancing analytical efficiency in pharmaceutical drug analysis.
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BibTeX
@article{dinc2026icrfassisted,
title = {{ICRF-assisted Box–Behnken design and optimization for rapid UPLC–PDA determination of dorzolamide hydrochloride and timolol maleate in an ophthalmic preparation}},
author = {Erdal Dinç and Eda Büker},
journal = {Journal of AOAC International},
year = {2026},
month = jun,
doi = {10.1093/jaoacint/qsag060},
url = {https://doi.org/10.1093/jaoacint/qsag060},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - ICRF-assisted Box–Behnken design and optimization for rapid UPLC–PDA determination of dorzolamide hydrochloride and timolol maleate in an ophthalmic preparation
AU - Erdal Dinç
AU - Eda Büker
PY - 2026
DA - 2026/06/25
JO - Journal of AOAC International
DO - 10.1093/jaoacint/qsag060
UR - https://doi.org/10.1093/jaoacint/qsag060
AB - Background: 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. Objective: This strategy describes the development of a rapid, chemometrically optimized Ultra Performance Liquid Chromatography coupled with a Photodiode Array Detector (UPLC–PDA) method for the simultaneous determination of dorzolamide hydrochloride (DH) and timolol maleate (TI) in a commercial ophthalmic preparation. Methods: A Box–Behnken experimental design and optimization approach was employed in combination with an 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. Results: Under the optimized conditions, complete chromatographic separation was achieved within 3 min using a BEH C18 column and a mobile phase consisting of acetonitrile–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 LODs 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. Conclusion: The proposed ICRF-assisted optimization approach provided high-resolution separation within minimal runtime and was successfully applied to the routine QC analysis of a commercial ophthalmic formulation. Highlights: The study demonstrates the effectiveness of composite response-based chemometric optimization in enhancing analytical efficiency in pharmaceutical drug analysis.
SN - 1060-3271
SN - 1944-7922
ER -