LC determination of the anti-ischemic and anti-hypertensive agent CDRI-93/478 in rat serum

CDRI-93/478 is a potent anti-ischemic and anti-hypertensive agent. This compound is in advanced stage of pre-clinical trials. A high-performance liquid chromatographic (HPLC) method was developed for the analysis of CDRI-93/478 in rat serum, a species used for safety evaluation. The HPLC analysis, applicable to 1 ml volumes of serum, involved double extraction of serum samples with diethyl ether at alkaline pH followed by separation on a spheri-5 cyano column and the use of fluorescence detector at excitation wavelength 250 nm and emission wavelength 372 nm. The method was sensitive with a limit of quantitation of 10 ng ml(-1) in rat serum and the recovery was more than 84%. The linearity was satisfactory as indicated by correlation of >0.99, in addition to the visual examination of the calibration curves. The precision and accuracy were acceptable as indicated by relative standard deviation (R.S.D.) ranging from 1.73 to 9.51%, bias values ranging from -7.31 to 8.68%. Moreover, CDRI-93/478 was stable in rat serum after being subjected to three freeze-thaw cycles. In-process stability evaluation showed the stability of the compound in processed samples lasted up to 168 h. The assay was found to be sensitive, specific, accurate, precise, and reliable for use in pharmacokinetic or toxicokinetic studies.