RP-HPLC Single Advance Method with Validation Studies for Imipramine Hydrochloride and its Intermediates
Indian Journal of Pharmaceutical Education and Research
Abstract
Background: A quantifiable method has been developed to determine the key starting material, its intermediates and known impurities in the presence of Imipramine Hydrochloride in the final API of a synthetic laboratory sample. This newly developed Reverse-Phase High-Performance Liquid Chromatography (RP-HPLC) method is, facile, specific and reliably practical. Materials and Methods: The separation column employed was Inertsil ODS-3 C18 with a mobile phase comprised of (A) 0.1% OPA (pH adjusted to 3.2) and (B) acetonitrile. Mobile phase A is a 100% buffer solution. Acetonitrile was used as mobile phase “B” and mobile phase B was used as 100% organic Solvent. The analytes were detected at 220 nm using a UV detector where the flow rate of the mobile phase was kept at 1.0 mL min-1 and the gradient program was set as T/% B: 0/30, 5/30, 10/80, 12/80, 15/30, 20/30 with a fixed flow rate of 1.0 mL min-1. Results: According to the regulatory standards advised by the ICH, the performance of this method is best agreed upon by all the important parameters. The approach used in the present work can be used for process development and determining the purity of associated compounds of Imipramine Hydrochloride, key starting material (2,2-dinitro-1,2-diphenylene ethane), intermediate-1 (2-2-diamino-1,2-diphenyl ethane diphosphate) and intermediate-2 (iminodibenzyl) all in a single method. Conclusion: This newly developed and validated method will save time and create ease by preventing the development of different methods for analyzing intermediates and impurity profiling.
Keywords
- Imipramine Hydrochloride
- API
- RP-HPLC
- UV detector
- Chromatogram
- Validation