Development and validation of RP-HPLC method for determination of amoxicillin residues and application to NICOMAC coating machine

Abstract

The Cleaning Validation protocol plays an important role in the field of pharmaceutical industries; its main task is the verification of cleaning procedures to ensure that complete removal of product residues, degradation products, preservatives, excipients, cleaning agents and cross-contamination of the previous active ingredients. A new RP-HPLC method is evaluated for determination of Amoxicillin (AMO) residues in NICOMAC coating machine using Betabasic-C18 (4.6mm x 250mm) 5µm or equivalent, mobile phase of a mixture of 0.05M sodium dihydrogen phosphate: methanol (95:5v/v) adjusted to pH 4.4 with orthophosphoric acid at a flow rate of 1.5mL/min, injection volume 100μL and UV detection at 230nm. The retention time of AMO is 6.292 min and the total run time is 7.0 min. A Linear relationship is obtained in the range 0.03 to 6 ppm with a correlation coefficient of 0.9989, limit of detection 0.05μgmL-1 and limit of quantitation of 0.15μgmL-1. The overall recovery is 100±15%; the relative standard deviation for precision and intraday precision is less than 2.0 %. The validation of the method is performed according to ICH guidelines and USP requirements for new methods, which include accuracy, precision, specificity, LOD, LOQ, robustness, ruggedness, linearity and range.