DEVELOPMENT AND VALIDATION OF A METHOD FOR QUANTIFICATION OF SIPONIMOD IN SPIKED RAT PLASMA

The creation of reliable bioanalytical LC-MS (liquid chromatography-mass spectroscopy) methods is crucial for the discovery, development, and eventual marketing approval of new drugs. A brand-new medication called siponimod was developed to treat Multiple Sclerosis (MS). This medication is a sphingosine-1-phosphate (S1P) receptor modulator and is hypothesised to contribute to the reduction of MS-related inflammation of the central nervous system. Since the last several decades, the usage of ant sclerosis agents has grown dramatically, however there is currently no bio analytical approach available for quantifying siponimod in biological matrix investigations employing LC-MS/MS. To create a novel, quick, and accurate LC-MS/MS approach for simultaneously measuring siponimod in rat plasma utilising the liquid-liquid extraction method (LLE) and deuterated siponimod (SId6). Method: Chromatographic separation was performed in isocratic mode using acetonitrile and OPA buffer at a flow rate of 1 ml/min on a reverse phase Phenyl C18 (150 mm x 4.6 mm, 3.5 m) column. Utilising a positive-mode electrospray ion interface and multiple reaction monitoring (MRM) procedures, quantification was accomplished.  Over the concentration range of 5.00–100.00pg/mL, the technique demonstrated good linearity. For siponimod at HQC and LQC, the intra-batch and inter-batch precision (%CV) was 4.3%, and the matrix effect (%CV) was 0.021% and 0.13%, respectively. The method’s ease of use makes it suitable for use in laboratories and offers a useful tool for pharmacokinetic investigations. The specific test has been expertly used in research with rats.