Abstract

The goal of the study was to develop a high throughput quantification method with precision and accuracy in plasma and brain matrices for Rivastigmine (RST). For the method development, a risk assessment was performed to first select the critical variables/factors and hence select the critical method parameters which were introduced in Design of Experiment to obtain the optimized analytical method. Protein precipitation and PLC High Performance Liquid Chromatography (HPLC)/ Ultraviolet (UV) were selected as the techniques for the drug extraction and estimation respectively. Further on validation, the method was selective and showed good linearity with recovery ranging between 96.73%–109.81% in plasma and 93.42%–108.39% in the brain. The method validation showed acceptable precision and accuracy and was found to be stable in both the matrices. The method sensitivity in both matrices was demonstrated by the lowest concentration detection at 75 ng.ml−1. Up until now, no HPLC-UV method is available with protein precipitation drug extraction for bioanalysis of RST in plasma and brain matrices. Using this validated method, nasal pharmacokinetics was carried out in Sprague Dawley rats to confirm the method applicability for RST quantification in biological matrices.

Key words: HPLC, Bioanalytical, Quality-By-Design, Rivastigmine, Pharmacokinetics