Title:QbD Assisted Development and Validation of UV Spectroscopic Method in
Estimation of Silymarin
Volume: 21
Issue: 5
Author(s): Ashwini Kumar Mishra, S.L. Neha, Laxmi Rani, Hitesh Kumar Dewangan and Pravat Kumar Sahoo*
Affiliation:
- Department of Pharmaceutics, Delhi Institute of Pharmaceutical Sciences and Research, Delhi Pharmaceutical Sciences
and Research University, New Delhi, 110017, India
Keywords:
Hepatotoxicity, analytical quality by design (AQbD), silymarin, nano formulation, design of experiment, design expert.
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.
Aims: 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.
