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As part of this study, sustainability assessment tools were used to assess the environmental impact and practicality of an innovative, straightforward RP\u2010UPLC method to analyze cyclopentolate (CLO) and its organic impurities simultaneously in pure and ophthalmic solutions at the same time. An optimization strategy based on Box\u2013Behnken design was employed to minimize experimental runs while optimizing chromatographic conditions. Using this design, four critical variables were evaluated comprehensively\u2014ethanol percentage in the mobile phase, pH, column temperature, and flow rate\u2014on chromatographic responses such as retention time, resolution between CLO and impurity, and theoretical plate count. As a result of desirable and overlay plots, an optimal condition was selected: 65:25, v\/v, ethanol and buffer, pH 4.25, 0.3\u2009mL\/min flow rate, and 4\u00b0C and 25\u00b0C sample and column oven temperatures, respectively, and the main peak retained for a little more than 3\u2009min. The calibration curves for CLO and impurities at concentrations from 5 to 50\u2009\u03bcg\/mL and 1 to 20\u2009\u03bcg\/mL showed a correlation value of 0.9998. Recoveries are \u00b115% of the actual amounts, which is acceptable. RP\u2010UPLC has been extensively designed for the coincidental estimation of anticholinergic drugs and their impurities. A combination of white and green tools was used to assess the method\u2019s environmental impact. ICH guidelines have been followed to validate the suggested strategy. This approach offers a reliable, fast, and eco\u2010friendly solution for routine pharmaceutical quality control of anticholinergic agents.<\/jats:p>","DOI":"10.1155\/ianc\/8827373","type":"journal-article","created":{"date-parts":[[2025,9,16]],"date-time":"2025-09-16T16:20:47Z","timestamp":1758039647000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Green RP\u2010UPLC Method for Simultaneous Determination of Cyclopentolate and Organic Impurities Using DoE and Sustainability Metrics"],"prefix":"10.1155","volume":"2025","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8648-6926","authenticated-orcid":false,"given":"Bandar R.","family":"Alsehli","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-2591-0946","authenticated-orcid":false,"given":"Abdullah 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