Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment

Abstract Intestinal coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses for the industry. It compromises the nutrition absorption, leading to weight loss and elevated mortality rates. Furthermore, the stress caused by the infection can compromis...

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Main Authors: Michel Y. Fares, Shimaa Ebrahim Abdel Aziz, Israa Abdelghafar khalil, Asmaa Othman El Demerdash, Neven M. Habib
Format: Article
Language:English
Published: Nature Portfolio 2025-02-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-024-84849-4
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author Michel Y. Fares
Shimaa Ebrahim Abdel Aziz
Israa Abdelghafar khalil
Asmaa Othman El Demerdash
Neven M. Habib
author_facet Michel Y. Fares
Shimaa Ebrahim Abdel Aziz
Israa Abdelghafar khalil
Asmaa Othman El Demerdash
Neven M. Habib
author_sort Michel Y. Fares
collection DOAJ
description Abstract Intestinal coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses for the industry. It compromises the nutrition absorption, leading to weight loss and elevated mortality rates. Furthermore, the stress caused by the infection can compromise the immune system, making poultry more susceptible to secondary infections and reducing overall productivity. As a result, simple analytical techniques are critical for determining anticoccidial drugs. A new, sensitive, and environmentally friendly HPLC method was developed for determining amprolium (AMP), sulfaquinoxaline (SUL), diaveridine (DIV), and vitamin K3 (VIT K3) in their formulations for the first time. Stability tests were performed under diverse stress conditions to verify the safety and efficiency of the formulation throughout its designated shelf time. These investigations ascertain the influence of various environmental conditions on a drug’s chemical stability and physical characteristics. A Supelcosil C18 column was used as the stationary phase, and 0.05 M KH2PO4 and acetonitrile were mixed in a ratio of 80:20 (v/v) as the developing system with a flow rate of 2.0 mL min−1. The proposed drugs were quantified at 260 nm. It was tested and found that the novel analytical method was linear for AMP and SUL between 20.0 and 60.0 µg mL−1, 2.0–6.0 µg mL−1 for VIT K3, and 2.1–6.3 µg mL−1 for DIV. The anticipated method was validated according to ICH guidelines. Advanced evaluation tools, such as GAPI, Red Green Blue (RGB 12, whiteness), Blue Applicability Grade Index (BAGI), the Analytical Eco-Scale, and (AGREE) assessed the sustainability profile of the proposed method, illustrating its enhanced environmental friendliness and sustainability.
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publishDate 2025-02-01
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spelling doaj-art-2e8ac51abcdb4019890e47290ce81fba2025-08-20T04:02:46ZengNature PortfolioScientific Reports2045-23222025-02-0115111610.1038/s41598-024-84849-4Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessmentMichel Y. Fares0Shimaa Ebrahim Abdel Aziz1Israa Abdelghafar khalil2Asmaa Othman El Demerdash3Neven M. Habib4Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Nahda University in Beni-Suef (NUB)Department of Analytical Chemistry, Faculty of Pharmacy (Girls), Al-Azhar UniversityDepartment of Analytical Chemistry, Faculty of Pharmacy (Girls), Al-Azhar UniversityDepartment of Analytical Chemistry, Faculty of Pharmacy (Girls), Al-Azhar UniversityDepartment of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Beni-Suef UniversityAbstract Intestinal coccidiosis is a significant parasitic disease affecting poultry, resulting in substantial economic losses for the industry. It compromises the nutrition absorption, leading to weight loss and elevated mortality rates. Furthermore, the stress caused by the infection can compromise the immune system, making poultry more susceptible to secondary infections and reducing overall productivity. As a result, simple analytical techniques are critical for determining anticoccidial drugs. A new, sensitive, and environmentally friendly HPLC method was developed for determining amprolium (AMP), sulfaquinoxaline (SUL), diaveridine (DIV), and vitamin K3 (VIT K3) in their formulations for the first time. Stability tests were performed under diverse stress conditions to verify the safety and efficiency of the formulation throughout its designated shelf time. These investigations ascertain the influence of various environmental conditions on a drug’s chemical stability and physical characteristics. A Supelcosil C18 column was used as the stationary phase, and 0.05 M KH2PO4 and acetonitrile were mixed in a ratio of 80:20 (v/v) as the developing system with a flow rate of 2.0 mL min−1. The proposed drugs were quantified at 260 nm. It was tested and found that the novel analytical method was linear for AMP and SUL between 20.0 and 60.0 µg mL−1, 2.0–6.0 µg mL−1 for VIT K3, and 2.1–6.3 µg mL−1 for DIV. The anticipated method was validated according to ICH guidelines. Advanced evaluation tools, such as GAPI, Red Green Blue (RGB 12, whiteness), Blue Applicability Grade Index (BAGI), the Analytical Eco-Scale, and (AGREE) assessed the sustainability profile of the proposed method, illustrating its enhanced environmental friendliness and sustainability.https://doi.org/10.1038/s41598-024-84849-4AmproliumSulfaquinoxalineDiaveridineVitamin K3HPLCGreenness assessment
spellingShingle Michel Y. Fares
Shimaa Ebrahim Abdel Aziz
Israa Abdelghafar khalil
Asmaa Othman El Demerdash
Neven M. Habib
Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment
Scientific Reports
Amprolium
Sulfaquinoxaline
Diaveridine
Vitamin K3
HPLC
Greenness assessment
title Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment
title_full Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment
title_fullStr Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment
title_full_unstemmed Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment
title_short Development and validation of a stability-indicating HPLC method for the simultaneous determination of anticoccidial drugs in veterinary formulations: greenness and whiteness assessment
title_sort development and validation of a stability indicating hplc method for the simultaneous determination of anticoccidial drugs in veterinary formulations greenness and whiteness assessment
topic Amprolium
Sulfaquinoxaline
Diaveridine
Vitamin K3
HPLC
Greenness assessment
url https://doi.org/10.1038/s41598-024-84849-4
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