A stability-illustrating HPLC-DAD method for assessment of two veterinary anti-parasitic drugs: appraisal of the method’s greenness and blueness

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Abstract

This paper represents an effective and reliable high-performance liquid chromatography-diode array detector (HPLC-DAD) method for the regular assay of Clorsulon (CLR) and Moxidectin (MOX) anti-parasitic drugs in injection solution and pure powder without derivatization processes. The mobile phase was composed of acetonitrile: methanol: water: acetic acid (56.0: 36.0: 7.5: 0.5 by volume). Besides, a Supelcosil C18 (4.60 mm ×15.0 cm, 5.0 μm) column was selected for completing the separation and quantitation of the two aforementioned veterinary drugs at a wavelength of 254 nm. The flow rate was set at 2.0 mL min−1 at the isocratic approach. We have conducted the degradation experiments using the HPLC-DAD instrument, adhering to the guidelines of the International Conference of Harmonization (ICH), subjecting CLR and MOX to light, heat, basic, acidic, and oxidative stressful conditions to figure out the ideal storage conditions and the possible medications that can be co-administered with them. CLR and MOX were quantified linearly from 400.0 to 1200.0 and from 40.0 to 120.0 µg mL−1, respectively. The maximum recorded degradation results were in acidic, basic, and oxidative conditions. Therefore, strong basic or acidic medications and oxidants shouldn’t be combined with CLR and MOX in a co-formulated medication. Greenness, carbon footprint, and blueness assessments for the novel method were conducted to verify the sustainability and functionality. The thirteen subdivisions in the GAPI pictogram, which were categorized as either green or yellow, refer to moderate green aspects. The final AGREE score of 0.56 and the majority of its subdivisions, ranging from dark green to yellow, indicated a relatively moderate level of greenness. This was primarily due to the significant acetonitrile content (56%) in the mobile phase. Using the HEXAGON method, the ultimate score is 0 out of 5 since the total calculated carbon footprint is less than 0.10. An eco-friendly method is one with a reduced carbon footprint score. The innovative HPLC method’s functioning and utility are indicated via its overall BAGI score of 80.0. Generally, the outcomes of the AGREE and GAPI pictograms indicate that the HPLC-DAD has a greenness feature, despite its moderate sensitivity.

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Elsayed, M. A., Algethami, F. K., Alotaibi, A. N., AlSalem, H. S., Mohammed Abd Elhalim, L., & Gamal, M. (2025). A stability-illustrating HPLC-DAD method for assessment of two veterinary anti-parasitic drugs: appraisal of the method’s greenness and blueness. BMC Chemistry, 19(1). https://doi.org/10.1186/s13065-025-01401-9

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