RP-HPLC METHOD DEVELOPMENT AND VALIDATION FOR THE ESTIMATION OF NELARABINE IN BULK AND IN INJECTION

Abstract

An innovative, cost-effective reverse phase high performance liquid chromatography (RPHPLC) approach has been devised to accurately and precisely determine the quantity of Nelaribine in both bulk and sterile dosage forms. The separation process was conducted using an Inertsil C18-ODS 3V column with dimensions of 250×4.6 mm and a particle size of 5 µm. Isocratic mode was employed, with a mobile phase consisting of a mixture of acetic acid pH4.0 buffer and methanol in a ratio of 75:25 (v/v). The mobile phase was pumped into the column at a flow rate of 1.5 mL min-1. The eluent from the column was detected using a UV
detector set at 260 nm. The whole duration of the experiment was 8 minutes, during which the column was kept at the ambient temperature. The observed retention time for Nelaribine was determined to be 3.99 minutes. The linearity of the standard curves was seen within the concentration range of 20-100 µg/ml, exhibiting a high coefficient of determination (R2 = 0.9998). The results of the experiment indicated a percentage recovery ranging from 100.0272% to 101.8%. Additionally, the relative standard deviation (RSD) was determined to be 0.3693%. The measured percentage content of a commercially available Nelaribine formulation was determined to be 100.20 %. The methodology was verified in accordance with the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) recommendations. The suggested RP-HPLC technique has been validated by investigations, which have shown that it has characteristics of simplicity, specificity, rapidity, reliability, and reproducibility. Therefore, the approach
presented in this study may be used for the regular analysis of Nelaribine in both bulk and injectable dose forms, specifically for quality control purposes.