Eco-friendly micellar HPLC approach for assessment of two anti-asthmatic breezhalers; Application of central composite design

Abstract

Short-term breathing problems are brought on by asthma, a chronic lung disease. Mometasone furoate (MOF) and Indacaterol (IDT), are co-formulated alone or with Glycopyrronium (GLY) in the form of inhalation powder to control the symptoms of asthma. A novel green hybrid micellar liquid chromatographic method was developed to analyze the three mentioned medications. Contrary to routine analysis, Quality by Design methodology was applied to maximize the resolution with the fewest number of trials. Fractional factorial design was used to study the impact of three significant variables: sodium dodecyl sulphate (SDS) concentration, methanol %, and the mobile phase pH. Separation of the analytes was optimized by central composite design using a mobile phase consists of (191.49 mM) SDS and (10.00 mM) potassium dihydrogen phosphate buffer: methanol (77.50: 22.50 v/v), pH 3.00 on a reversed phase C18 column, within eight minutes. The flow rate was 1.2 mL/min, with 20 µL injection volume, and UV detection at 210 nm. Green Analytical Procedure Index, and Analytical Greenness Calculator were used for the method’s greenness evaluation, and it was deemed to be an effective green methodology. The developed approach was effectively applied for quantitative determination of the three drugs in bulk powder and two pharmaceutical dosage forms. International Conference of Harmonization requirements were adhered to when validating the approach. Under optimized conditions, calibration curves of the studied drugs were linear over the concentration range of (10.00 – 100.00 µg/mL) with high accuracy results and correlation coefficients of 0.9997 for MOF, 0.9996 for IDT and 0.9996 for GLY. Additionally, a precise approach has been demonstrated by the intra- and inter-day coefficients of variation ≤ 2%. The LOD and LOQ were (2.39 μg/mL) and (7.25 μg/mL) for MOF, (2.67 μg/mL) and (8.08 μg/mL) for IDT, and (2.80 μg/mL) and (8.48 μg/mL) for GLY, respectively.