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Updated: May 5, 2026

Qualitative and Quantitative Assays for Detection and Characterization of Protein Antimicrobials
Published on: April 10, 2016
Full green assay of parenteral dosage forms of polymyxins utilizing xanthene dye: application to content uniformity
Mahmoud A Abdelmajed1, Khalid M Badr El-Din2, Tamer Z Attia3
1Department of Pharmaceutical Analytical Chemistry, Faculty of Pharmacy, Deraya University, New Minia, Egypt. Mahmoud.alaa81@yahoo.com.
Abstract:
Due to the lack of other treatment options, a rebirth of polymyxins is urgently required. Colistin (also called polymyxin E) and polymyxin B are the only two examples of this antibiotic class that were effectively employed in such critical situations. In the present work, both of the two studied medications were quantified via a simple, green, and non-extracting spectrophotometric approach based on the formation of ion-pair complexes with Erythrosine B. Without using any organic solvents, the pink color of the created complexes was detected at wavelength = 558 nm. To achieve the highest intensity of absorbance, optimum conditions were established by the screening of many experimental factors such as pH, buffer volume, the volume of Erythrosine B, and the time consumed to undergo the reaction. For Colistin and Polymyxin B respectively, Beer-Lambert's law was observed at the concentration ranges of 1-6, 1-9 µg mL- 1. The technique was approved and validated following ICH recommendations. Lastly, the suggested approach has been successfully implemented to quantify the cited medications colorimetrically, for the first time, in their parenteral dosage forms with excellent recoveries. Also, Content uniformity testing was implemented.
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