QBD-Based Development of Experimental Design with Response Surface-Based Optimization of Siponimod by Chromatographic and Spectroscopic Method

Background

A recent technique for characterizing Siponimod, a multiple sclerosis medication, was developed using Ultraviolet spectroscopic analysis, high-performance liquid chromatography, high-performance Thin-Layer Chromatography and spectrofluorimetric analysis.

Purpose

To optimize the amount of Siponimod in Pharmaceutical formulation by parallel methods assisted with response surface design and to validate the method as per ICH guidelines.

Materials and Methods

The UV absorbance and the fluorescence intensity were separately measured using the Cary 5000 double-beam spectrophotometer and the Cary Eclipse Spectrofluorometer. The Agilent C18 column was used for chromatographic separation and the mobile phase comprised of phosphate buffer and Acetonitrile at different concentrations. Using Digital Expert Stat-Ease -360 software, the response surface approach was used to optimize the methods’ variables using the Box-Behnken design. HPTLC determination was carried out using a Camag Linomet densitometric scanner.

Results

Wavelength maximums of Siponimod detected at 258 nm, the medication shows a chromatographic peak that is distinct and easy to interpret. There was a fluorescence excitation at 486 nm. The recovery percentage varied from 92% to 100% w/w with a relative standard deviation of less than 2. By ICH regulations, the procedures were confirmed and a transitional deterioration pattern was perceived while taking into account the medications’ stability in a forced stress environment.

Conclusion

It was discovered that the procedure was reliable and suitable for regular screening of Siponimod in bulk dose form as well as pharmaceutical formulation.