Academic Journal
Validation and Greenness Evaluation of TLC and HPLC Methods for Analysis of Nitroxynil, Clorsulon, and Benzyl Alcohol in Flulock® Veterinary Injectable Formulation.
| Τίτλος: | Validation and Greenness Evaluation of TLC and HPLC Methods for Analysis of Nitroxynil, Clorsulon, and Benzyl Alcohol in Flulock® Veterinary Injectable Formulation. |
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| Συγγραφείς: | Farid NF; Beni-Suef University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department, Beni Suef, 62514, Egypt., Bahgat ES; Evaluation and Accreditation Department, Egyptian Drug Authority, 51 Ministry of Agriculture Street, El-Batal Ahmed Abd El-Aziz, Al Agouzah, Cairo, 12611, Egypt., Hegazy AM; Beni-Suef University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department, Beni Suef, 62514, Egypt., Abdelfatah RM; Beni-Suef University, Faculty of Pharmacy, Pharmaceutical Analytical Chemistry Department, Beni Suef, 62514, Egypt. |
| Πηγή: | Journal of AOAC International [J AOAC Int] 2026 Sep 01; Vol. 109 (5), pp. 665-672. |
| Τύπος έκδοσης: | Journal Article; Validation Study |
| Γλώσσα: | English |
| Στοιχεία περιοδικού: | Publisher: Oxford University Press Country of Publication: England NLM ID: 9215446 Publication Model: Print Cited Medium: Internet ISSN: 1944-7922 (Electronic) Linking ISSN: 10603271 NLM ISO Abbreviation: J AOAC Int Subsets: MEDLINE |
| Imprint Name(s): | Publication: 2020- : Oxford : Oxford University Press Original Publication: Arlington, VA : AOAC International, c1992- |
| Ιατρικοί όροι (MeSH): | Benzyl Alcohol*/analysis , Veterinary Drugs*/analysis , Dinitrobenzenes*/analysis, Chromatography, Thin Layer/methods ; Chromatography, High Pressure Liquid/methods ; Green Chemistry Technology/methods ; Reproducibility of Results ; Animals ; Limit of Detection ; Injections |
| Περίληψη: | Background: 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. Objective: 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. Methods: TLC separation was performed on silica gel 60 F254 plates using a mobile phase of ethyl acetate-ammonia-methanol (9.5 + 0.5 + 0.05, by volume), with densitometric detection at 254 nm. The HPLC method employed a C8 column with gradient elution using 3 mM tetrabutylammonium hydrogen sulfate (pH 6.3) and a mixture of acetonitrile and methanol, with UV detection at 215 nm. 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. Results: The TLC method showed linearity over concentration ranges of 0.5-3.5 µg/band and 0.2-2 µg/band for NIT and CLO, respectively, with correlation coefficients of 0.9996 and 0.9994. The HPLC method exhibited linearity over ranges of 1-24 µg/mL and 1-21 µg/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. Conclusion: 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. Highlights: Simultaneous determination of NIT, CLO, and benzyl alcohol; validated TLC-densitometric and HPLC methods; successful application to Flulock® veterinary injectable formulation; high accuracy, precision, and selectivity; greenness evaluated using MoGAPI, AGREE, and BAGI tools. (© The Author(s) 2026. Published by Oxford University Press on behalf of AOAC INTERNATIONAL. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.) |
| Substance Nomenclature: | LKG8494WBH (Benzyl Alcohol) 0 (Veterinary Drugs) 0 (Dinitrobenzenes) |
| Entry Date(s): | Date Created: 20260411 Date Completed: 20260908 Latest Revision: 20260909 |
| Update Code: | 20260910 |
| DOI: | 10.1093/jaoacint/qsag028 |
| PMID: | 41965065 |
| Βάση Δεδομένων: | MEDLINE |
| ISSN: | 1944-7922 |
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| DOI: | 10.1093/jaoacint/qsag028 |