An Ecologically Sound RP-HPLC Method for Estimating Citicoline using AQbD with Degradation Studies

Abstract

Background: Citicoline is widely used for neurological complications. This research work was carried out using reverse-phase high-performance liquid chromatography (RP-HPLC) employing green analytical chemistry principles. Materials and Methods: The method was completely developed with ethanol:water (20:80 v/v) as the mobile phase, 0.1% orthophosphoric acid was used to adjust the pH to 3.5, and an Inertsil ODS 3 (250 x 4.6 mm, 5 μm) column as the stationary phase, the flow rate was set at 1-mL/min. The column temperature was maintained at 35ºC, and an injection volume of 10 μL was used for detection at a wavelength of 250 nm. Results and Discussion: The peak was eluted with isocratic elution with a retention time of 3.996 minutes. The linear concentration range for this method was 10 to 50 μg/mL, and the correlation coefficient R2 was 0.999. The forced degradation studies have shown that 3.5% w/w degraded in acid, 1.35% w/w in alkali, and 0.2% w/w in oxidation. The products formed during degradation were characterized using gas chromatography-mass spectrometry (GC-MS). Conclusion: The International Council for Harmonisation (ICH) guidelines were followed in the development and validation of the method optimized by the use of central composite design, and the method’s greenness was assessed through the use of AGREE, AES, GAPI, and AMGS, among other green assessment tools.