Category: Formulation and Quality
Purpose: A degradant impurity - 1,1-Dimethyl-3-Hydroxy-Pyrrolidinium Bromide is lack of chromophore in its structure, hence it does not shows any absorbance in UV-Visible range. The purpose of this work is to develop and validate a simple, accurate, selective and sensitive method for estimation of 1,1-Dimethyl-3-Hydroxy-Pyrrolidinium Bromide impurity in Glycopyrrolate Bromide by Liquid Chromatography–Tandem Mass Spectroscopy.
Methods: Impurity profiling is now receiving critical attention from regulatory authorities. For trace level quantification of potential degradation impurities, conventional analytical techniques like high-performance liquid chromatography (HPLC) and gas chromatography (GC) are inadequate; consequently, there is a great need to apply hyphenated analytical techniques to develop sensitive analytical methods for the analysis of pharmaceuticals.
A selective and sensitive liquid chromatography-tandem mass spectrometry (LC-MS) method was developed for the estimation of 1,1-Dimethyl-3-Hydroxy-Pyrrolidinium Bromide impurity in Glycopyrrolate Bromide. The LC-MS analysis was done on XBridge HILIC (100 mm × 4.6 mm, 5 μm) analytical column, and the mobile phase used was 10mM Ammonium Formate with 0.2% Formic acid as mobile phase-A and acetonitrile as mobile phase-B with a gradient programme of 5.0 minutes. The flow rate used was 1.2 mL/min. Triple quadrupole mass detector coupled to positive electrospray ionization operated in multiple reaction monitoring (MRM) mode was used for the quantification at m/z 116.10 ± 0.5. The method was validated as per International Conference on Harmonization (ICH) guidelines for specificity, precision, accuracy, linearity, range and limit of detection & limit of quantitation.
Results: The developed method was validated and validation studies revealed the method is specific, precise, accurate, linear, and robust. The impurity peak was eluted at about 3.2 minutes. Calibration plot was linear over the concentration range of 0.050 µg/mL to 2.000 µg/mL. RSD for system suitability was found 1.3 %. Limit of detection and limit of quantification was found 0.017 µg/mL and 0.051 µg/mL respectively. The high recovery and low relative standard deviation confirm the suitability of the method. The developed method was applied to determine the 1,1-Dimethyl-3-Hydroxy-Pyrrolidinium Bromide impurity in Glycopyrrolate Bromide API and formulations.
Conclusion: The developed method was successfully applied for the quantitative determination of 1,1-Dimethyl-3-Hydroxy-Pyrrolidinium Bromide impurity in Glycopyrrolate Bromide API and formulations in quality control laboratory and could be used for routine analysis.