QbD Assisted Development and Validation of UV Spectroscopic Method in Estimation of Silymarin

Abstract

Background: Silymarin is a flavonoid utilised in liver dysfunction for years; new studies have emphasised its potential utility as a therapy for nanoparticulate targeting in many other disorders. In order to establish the product quality of such compounds, researchers have been trying to develop a robust method of analysis, but the methods developed till now are too expensive and time-consuming. Here we proposed quality by design-assisted development and validation of the UV spectroscopic method using Design-Expert® software in the estimation of Silymarin for fabrication of nanoparticulate formulations, which is simple, accurate, cost-effective and non-tedious. Aim: The proposed method is a simple, new, robust, accurate, and precise UV visible spectroscopic technique for estimating silymarin produced in nano-formulations utilising an analytical quality by design (AQbD) approach. Methods: A UV spectrophotometric technique was established, as maximum absorption (287.7) was measured using a Shimadzu UV-1800 double-beam UV visible spectrophotometer. The characterization of silymarin was done by melting point, DSC and FTIR techniques. The two critical method variables chosen were scanning speed and sample interval to be analysed by the design of experiment methodology utilizing the central composite design principle, which shows robustness and optimized technique involved in this work. Results: The spectroscopy technique was developed and validated as per International Conference of Harmonization recommendations. The Beer's-Lambert rule was followed in a series of 2-12 μg/ml dilution increments, with a correlation value of R2 = 0.999. The method's linearity was shown to be excellent across the concentration range. The percent recovery of the current method approach was determined to be within the confidential limitations, i.e., less than 2% expressed as % RSD, and the methodology was proved to be precise at inter and intraday variations (% RSD). The LOD and LOQ were found to be 0.264 μg/ml and 0.801 μg/ml which were also determined correctly. During specificity testing, no interfering peaks were found. Conclusion: This UV approach has been used successfully to determine the quantity of silymarin present in the nanoparticulate formulation, which can be used for testing its other pharmaceutical dosage forms.