Stability Indicating Assay Method Development and Validation by RP-UPLC with PDA detector for Simultaneous Estimation of Glycopyrrolate and Neostigmine in Pharmaceutical dosage form

Abstract

The presented research aimed to create a simple, precise, and accurate isocratic reversed-phase stability indicating Ultra Performance Liquid Chromatography (UPLC) method for simultaneous quantification of Glycopyrrolate and Neostigmine in bulk and injectable dosage form. On a Symmetry-C18(150mm X 4.6mm, 3.5m) column, isocratic separation was accomplished. The mobile phase is composed of acetonitrile and 1% orthophosphoric acid buffer (70:30 v/v) flowing at a rate of 1ml/min, with detection at 255nm utilising a photo-diode array detector. To apply stress conditions, the drug was treated to acid degradation, alkali degradation, peroxide degradation, photolysis, and heat. Specificity, precision, linearity, accuracy robustness, and solution stability were all validated. Glycopyrrolate and Neostigmine showed linearity in the range of 1-15mg mL-1 and 5-75mg mL-1, respectively. Glycopyrrolate accuracy was from 98.9 to 100.2 percent, whereas Neostigmine accuracy ranged from 98.4 to 100.2 percent. The detection of Glycopyrrolate and Neostigmine test does not interfere with degradation products produced as a result of stress studies, hence this method is regarded as stability-indicating.