Development and Validation of a HPLC Method for MS-153 Quantification: Assessment of its Stability in Rat Plasma and Brain Homogenate

Abstract

MS-153 is a novel pyrazoline compound that serves as a potential neuroprotective therapeutic agent during ischemia. Development of a convenient, quick, and robust analytical method to quantify MS-153 in biological samples is necessary to understand it’s in vivo pharmacokinetic/pharmacodynamics (PKPD) profiles. An isocratic reverse-phase HPLC method was developed and validated for quantification of MS-153. Chromatographic separation was achieved with a C18 column. Mobile phase consisting of water/acetonitrile (85/15, v/v) was pumped at a flow rate of 1.0 mL/min. The retention time of MS-153 (λmax=260 nm) was found to be 7.15 minutes. A calibration curve established over a range of 0.78125 ng to 500 ng showed a correlation coefficient of 1.0. The LOD and LOQ were found to be 0.164 and 0.496 ng, respectively. The accuracy, intra-day precision, and inter-day precision were found to be 99.97% to 101.66% (recovery), 0.21% to 0.55% (RSD), and 0.32% to 0.82% (RSD), respectively. MS-153 was analysed from biological samples by adding methanol to remove proteins in the biological matrix prior to HPLC analysis. The extraction efficiency was found to be 100%. The developed method was also used to analyse the stability of MS-153 in diluted blank rat plasma and brain homogenate samples. Results indicated that no significant degradation of MS-153 was observed at 37°C for 6 h.