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Two different designs including fractional factorial and Box\u2013Behnken designs were implemented for screening and optimization steps, respectively. The optimum chromatographic analysis was achieved using a mixture of isopropanol and 20\u00a0mM potassium dihydrogen phosphate solution (pH 3.0) in the ratio 27:73 as mobile phase. The flow rate was 1.5\u00a0mL\/min and column oven temperature was 40\u00a0\u00b0C. Chromatographic analysis was performed on Eclipse plus C18 (100\u00a0mm\u2009\u00d7\u20094.6\u00a0mm\u2009\u00d7\u20093.5\u00a0\u03bcm) column with DAD detector set at 220\u00a0nm. A linear response was acquired over the range of 2.5\u201360\u00a0\u03bcg\/mL and 1\u201350\u00a0\u03bcg\/mL for benoxinate and fluorescein respectively. Stress degradation studies were executed under acidic, basic, and oxidative stress conditions. The method was implemented for quantitation of cited drugs in ophthalmic solution with mean percent recovery\u2009\u00b1\u2009SD of 99.21\u2009\u00b1\u20090.74 and 99.88\u2009\u00b1\u20090.58 for benoxinate and fluorescein respectively. The proposed method is more rapid and eco-friendly compared to the reported chromatographic methods for determination of cited drugs.<\/jats:p>","DOI":"10.1038\/s41598-023-37548-5","type":"journal-article","created":{"date-parts":[[2023,6,28]],"date-time":"2023-06-28T10:01:59Z","timestamp":1687946519000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Quality by design paradigm for optimization of green stability indicating HPLC method for concomitant determination of fluorescein and benoxinate"],"prefix":"10.1038","volume":"13","author":[{"given":"Amira H.","family":"Kamal","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ahmed A.","family":"Habib","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sherin F.","family":"Hammad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Safa M.","family":"Megahed","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2023,6,28]]},"reference":[{"key":"37548_CR1","doi-asserted-by":"publisher","first-page":"369","DOI":"10.1039\/D0AY02169E","volume":"13","author":"M Gamal","year":"2021","unstructured":"Gamal, M., Naguib, I. 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