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Implementation of different separation techniques for resolving ternary mixture of Paracetamol, Pseudoephedrine Hydrochloride and Chlorpheniramine maleate with further quantification

Farid Joliana F., Mostafa Nadia M., Essam Hebatallah M., Fayez Yasmin M.

Acta Chromatographica

2022

Vol. 34, no. 3

351--360

Due to the wide applicability of separation techniques that rely on the property of differential migration in pharmaceutical formulations analysis, different analytical strategies have been proposed to resolve mixtures of multi-components pharmaceuticals. Three separation methods were developed and validated for the simultaneous determination of Paracetamol (PAR), Pseudoephedrine HCl (PSE) and Chlorpheniramine maleate (CHP). The first method is a thin-layer chromatographic (TLC) separation, followed by densitometric measurement. The separation was carried out on aluminium sheet of silica gel 60 F254 using ethanol:chloroform:ammonia (1:7:0.4, by volume) as the mobile phase. Determination of PAR, PSE and CHP was successfully applied over the concentration ranges of 3–25 µg/band, 0.5–10 µg/band and 0.1–6 µg/band, respectively. The second method is HPLC separation that was achieved on C18 column using the mobile phase acetonitrile:phosphate buffer pH 5 (10:90, v/v) at a flow rate 1 mL min⁻¹. PAR, PSE and CHP were determined by HPLC in concentration ranges of 5–400 μg mL⁻¹, 2–40 μg mL⁻¹ and 0.5–16 μg mL⁻¹, respectively. The third method is a capillary electrophoresis (CE) separation. The electrophoretic separation was achieved using 20 mM phosphate buffer (pH 6.5) at 20 kV. The linearity was reached over concentration ranges of 30–250 μg mL⁻¹, 5–50 μg mL⁻¹ and 0.8–20 μg mL⁻¹ for PAR, PSE and CHP, respectively. The developed methods were validated with respect to linearity, precision, accuracy and system suitability. The proposed methods were successfully applied for bulk powder and dosage form analysis with RSD of precision <2%. Moreover, statistical comparison with the official methods confirms the methods' validity.

Optimization and validation of Eco-friendly RP-HPLC and univariate spectrophotometric methods for the simultaneous determination of Fluorometholone and Tetrahydrozoline hydrochloride

Essam Hebatallah M., Saad Martin N., Elzanfaly Eman S., Amer Sawsan M.

Acta Chromatographica

2021

Vol. 33, no. 3

216--227

A sensitive RP-HPLC method is presented for the simultaneous quantification of Fluorometholone (FLM) and Tetrahydrozoline hydrochloride (THZ). The method has the advantages of being rapid, accurate, reproducible, ecologically acceptable and sensitive. The separation utilized C8 Xbridge® column and mobile phase mixture of Acetonitrile/phosphate buffer pH 3 ± 0.1 (70:30, v/v) with UV detection at 230 nm. Stepwise optimization and factors affecting separation are properly discussed. Different factors were optimized such as stationary phase, selection of organic solvent and its content, buffer pH and concentration, flow rate, elution type and detection wavelength. The studied drugs were efficiently separated in 3.4 min with high resolution. Also, two univariate spectrophotometric methods have been optimized for the quantification of the studied drugs. Method 1: dual wavelength for THZ and iso-absorptive point for FLM, Method 2: ratio difference (RD) for THZ and first derivative FLM utilizing methanol as a solvent. These methods are accurate, precise with minimal data manipulation. Greenness of the methods was estimated using eco-scale tool where the presented methods were found to be excellent green with eco-score of 83 for HPLC and 80 for spectrophotometry. The methods are validated in conformance with ICH guidelines, with acceptable accuracy, precision, and selectivity. The suggested methods can be employed for the economic analysis of THZ and FLM in their pure form and binary ophthalmic formulation, that can be employed by quality control laboratories.