{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,9]],"date-time":"2026-09-09T20:14:04Z","timestamp":1788984844281,"version":"build-2803163510"},"reference-count":38,"publisher":"Oxford University Press (OUP)","issue":"5","license":[{"start":{"date-parts":[[2026,4,8]],"date-time":"2026-04-08T00:00:00Z","timestamp":1775606400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Nitroxynil (NIT) and clorsulon (CLO) are commonly co-formulated in veterinary injectable formulations, where benzyl alcohol is used as a preservative, necessitating selective and reliable analytical methods for their simultaneous determination.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>This study aimed to develop and validate sensitive, selective, and eco-friendly TLC and HPLC methods for the simultaneous determination of NIT, CLO, and benzyl alcohol in their pharmaceutical formulation.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>TLC separation was performed on silica gel 60 F254 plates using a mobile phase of ethyl acetate\u2013ammonia\u2013methanol (9.5 + 0.5 + 0.05, by volume), with densitometric detection at 254\u2009nm. The HPLC method employed a C8 column with gradient elution using 3\u2009mM tetrabutylammonium hydrogen sulfate (pH 6.3) and a mixture of acetonitrile and methanol, with UV detection at 215\u2009nm. Method validation was conducted in accordance with International Conference on Harmonization (ICH) guidelines. Greenness assessment was performed using Modified Green Analytical Procedure Index (MoGAPI), Analytical Greenness Metric (AGREE), and Blue Applicability Grade Index (BAGI) tools.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>The TLC method showed linearity over concentration ranges of 0.5\u20133.5\u2009\u00b5g\/band and 0.2\u20132\u2009\u00b5g\/band for NIT and CLO, respectively, with correlation coefficients of 0.9996 and 0.9994. The HPLC method exhibited linearity over ranges of 1\u201324\u2009\u00b5g\/mL and 1\u201321\u2009\u00b5g\/mL for the respective drugs. Both methods demonstrated high precision, accuracy, selectivity, and reproducibility, enabling effective separation of the analytes in the presence of benzyl alcohol. Greenness evaluation confirmed the low environmental impact of the proposed methods.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>The developed TLC and HPLC methods are reliable, accurate, and environmentally sustainable for the simultaneous determination of NIT, CLO, and benzyl alcohol in veterinary injectable formulations.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Highlights<\/jats:title>\n                    <jats:p>Simultaneous determination of NIT, CLO, and benzyl alcohol; validated TLC\u2013densitometric and HPLC methods; successful application to Flulock\u00ae veterinary injectable formulation; high accuracy, precision, and selectivity; greenness evaluated using MoGAPI, AGREE, and BAGI tools.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1093\/jaoacint\/qsag028","type":"journal-article","created":{"date-parts":[[2026,4,7]],"date-time":"2026-04-07T11:26:38Z","timestamp":1775561198000},"page":"665-672","source":"Crossref","is-referenced-by-count":0,"title":["Validation and Greenness Evaluation of TLC and HPLC Methods for Analysis of Nitroxynil, Clorsulon, and Benzyl Alcohol in Flulock\u00ae Veterinary Injectable Formulation"],"prefix":"10.1093","volume":"109","author":[{"given":"Nehal F","family":"Farid","sequence":"first","affiliation":[{"name":"Beni-Suef University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department , Beni Suef, 62514,","place":["Egypt"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-5025-4648","authenticated-orcid":false,"given":"Ebtehal S","family":"Bahgat","sequence":"additional","affiliation":[{"name":"Egyptian Drug Authority Evaluation and Accreditation Department, , 51 Ministry of Agriculture Street, El-Batal Ahmed Abd El-Aziz, Al Agouzah , Cairo, 12611,","place":["Egypt"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Amira M","family":"Hegazy","sequence":"additional","affiliation":[{"name":"Beni-Suef University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department , Beni Suef, 62514,","place":["Egypt"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0480-2118","authenticated-orcid":false,"given":"Rehab M","family":"Abdelfatah","sequence":"additional","affiliation":[{"name":"Beni-Suef University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department , Beni Suef, 62514,","place":["Egypt"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,4,8]]},"reference":[{"key":"2026090915261993900_qsag028-B1","first-page":"93","volume":"24","author":"Seelig","year":"2024","journal-title":"Int. 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