Novel Chromatographic Method Development and Validation for Nitroglycerin Determination in Semi-Solid Dosage Forms by RP-HPLC

The method was validated according to ICH Q2(R2) guidelines included the system suitability, specificity, linearity, accuracy, homogeneity, robustness and forced degradation. The report exhibits a robust and stability-indicating RP-HPLC method developed for the quantification of Nitroglycerin in 4 mg/g ointment formulations. Chromatographic separation was performed using an L1 packing C18 column (150 mm × 4.6 mm, 5 µm) with an isocratic mobile phase of water and methanol in a 60:40 (v/v) ratio. The flow rate was maintained at 1.0 mL/min, and detection was carried out using a UV detector set at 220 nm and run time was 15.0 min. It revealed excellent system suitability, specificity without interference from excipients, impurities, degradants, or matrix components and linearity over the range of 40–120 µg/mL. Percentage accuracy (recovery) values obtained for the NTG is from 99.3% to 100.1%. Photodiode array (3D-PDA) detector unveils homogeneity of the NTG peak under stress conditions confirmed uniform distribution across formulation layers. Robustness process confirmed method reliability, method transfer and reduced errors and reliability under varied conditions. Forced degradation study reveal specificity and its ability to distinguish NTG from its degradation products and metabolites, without interference under the various stress conditions such as acid and base hydrolysis, oxidation, thermal, humidity, and photolytic degradation. The assessment of greenness of analytical HPLC was carried out using Analytical Eco-Scale, AGREE, and GAPI. The Eco-Scale score of 87, the AGREE tool yielded a score of 0.63 and the GAPI pictogram revealed a balanced profile. Therefore, such worthwhile quantification study intended for regulatory compliance approach in the pharmaceutical and healthcare avenue.