Penalization and Color Code-Technical Approaches for Methods' Greenness and Whiteness Appraisal in Veterinary Medication: Assay of Toltrazuril Suspension.

Abstract

Intestinal coccidiosis is a debilitating disease in poultry and livestock, leading to economic impact worldwide. Coccidiosis is prevented and treated in broilers by the inclusion of anticoccidials in feed. Toltrazuril is administered in potable water to treat coccidiosis. Three robust analytical methods for quantitation of toltrazuril in pure and pharmaceutical formulations are developed. Furthermore, ecological metrics; either penalization- or color-code-based techniques are applied for the appraisal of assays. Firstly, Second-Derivative (Δλ; 5 nm) spectrophotometric method; Toltrazuril is measured from peak to peak at 244-260 nm within a linearity range of 5-25 μg/mL. The second one is a high-performance thin-layer chromatography (HPTLC) analysis performed on an aluminum sheet of silica gel using ethyl acetate, methanol, ammonium chloride buffer, and water (8:1:0.5:0.5) (%V/V) as the elution phase. Toltrazuril, at a retardation factor of 0.66 ± 0.01, is linearly determined in the range of 1-9 μg/spot at 243 nm. The third one is Reversed Phase-HPLC-diode array detection, using Agilent column C18 (5 μm, 4.6 x 150 mm) in isocratic elution mode with a mobile phase of acetonitrile and water in a ratio of 80:20 (v/v), respectively, at 1 mL/min flow rate. Toltrazuril elutes at a retention time of 2.58 ± 0.1 min and is linearly determined at 243 nm in the range of 0.25-25 μg/mL. Calculated 2D-values and peak areas are highly correlated to their corresponding drug concentrations at coefficients; r > 0.999. All methods were ICH validated and applied to dosage form with satisfactory % recoveries (97-103%). Statistical comparisons reported one using t-test and F-test disclose insignificant variation. Examining greenness and whiteness norms, proposed methods were evaluated and ranked alongside four different reported methods. The proposed methods are green, accurate, and can be applied in routine quality control for the determination of toltrazuril in pharmaceutical formulations.