Green Analytical Method Development and Validation of Rasagiline Mesylate by NP and RP HPTLC and RP-HPLC Methods

Abstract

Green chemistry principles were applied to develop a single process that achieves both environmental friendliness and robustness. Previous studies have reported various HPLC and HPTLC methods for evaluating Rasagiline Mesylate, but no green approach utilizing concepts like GAC (green analytical chemistry) has been described in the literature. In this study, we established and validated Green-NP-HPTLC and Green-RP-HPTLC techniques for the identification of Rasagiline Mesylate in drug substances and drug products. Rasagiline Mesylate is a monoamine oxidase inhibitor commonly used to treat Parkinson's disease symptoms. The Reverse Phase (RP) and Normal Phase (NP) HPTLC methods employed in this work enable precise measurement and verification of Rasagiline Mesylate in drug formulations. Analytical performance criteria, including linearity, accuracy, precision, specificity, limit of detection, limit of quantification, and robustness, were evaluated according to the ICH Q2R1 recommendations. In the NP-HPTLC method, chromatograms were generated using a precoated plate and a mobile phase consisting of butanol, ethyl acetate, and glacial acetic acid (4.5:4.5:1% v/v) on silica gel. Densitometric absorbance mode at 264 nm was used to determine the concentrations, and Rasagiline Mesylate exhibited an RF value of 0.32. The method showed excellent linearity (r2 = 0.999) within the linear dynamic range, with recovery values ranging from 99.78% to 99.98% and a low relative standard deviation (RSD) of 1.04%. For the RP-HPTLC method, chromatograms were obtained using an isopropanol: water mobile phase (6:4% v/v) on silica gel plates. Densitometric absorbance at 264 nm was employed for measurement. Rasagiline Mesylate exhibited an RF value of 0.69. The method demonstrated excellent linearity (r2 = 0.999) within the linear dynamic range, with recovery values ranging from 99.80% to 99.98% and a low RSD of 1.07%. Additionally, a green RP-HPLC method was developed and validated for the measurement of Rasagiline Mesylate. The separation was performed using a mobile phase of water and isopropanol (60:40% v/v) on a Phenomenex C18 column (250 mm x 4.6 mm, 5 μm). The eluentwas monitored at 264 nm, and the method exhibited excellent linearity (r2 > 0.9999) within the test limits. Reproducibility and intermediate precision were determined to be satisfactory. Comparing the three methods, Green-Reverse Phase-HPTLC demonstrated higher precision, speed, accuracy, and robustness for the identification of Rasagiline Mesylate when compared to Green-Normal Phase-HPTLC and Green-Reverse Phase-HPLC. The Analytical Greenness (AGREE) grades for Green-Normal Phase-HPTLC, Green-Reverse Phase-HPTLC, and Green Reverse Phase-HPLC were found to be 0.92, 0.87, and 0.85, respectively, indicating excellent greenness profiles."