High-throughput quantification of a novel thiazolidinedione MCC-555 in rat plasma by ultra-fast liquid chromatography and its application in pharmacokinetic studies.

MCC-555 is a novel thiazolidinedione which reduces plasma glucose concentrations in Type 2 diabetes mellitus models due to enhancement of insulin sensitivity. A highly sensitive and selective quantitative method to accurately determine MCC-555 in rat plasma is crucial to the success of pharmacokinetic studies of MCC-555. To this purpose we have developed and validated a high-throughput method in a 96-well plate format using ultra-fast liquid chromatography (Shimadzu Prominence UFLC system) for the determination of MCC-555 in rat plasma. MCC-555 along with the internal standard resveratrol was extracted from 50 microl of rat plasma by liquid-liquid extraction using ethyl acetate. Baseline separation of MCC-555 and resveratrol was achieved using UFLC technology on a C18 stationary-phase column with 2.2 microm particle size. The influences of flow rate, column temperature and mobile phase pH on chromatographic performance were investigated. Comparing to the conventional HPLC method, UFLC showed many advantages including reduced run time, less solvent consumption and increased sensitivity. The UFLC method was sensitive with a lower limit of quantification of 0.002 microg/ml, with good linearity (r>0.999) over the linear range of 0.002-2.0 microg/ml. The intra- and inter-run precision was less than 8.6% and accuracy ranged from -6.4 to 8.2% for quality control samples. The extraction recovery from plasma was no less than 80%. The validation and sample analysis results show that the method is precise, accurate and well suited to support pharmacokinetic studies in rats involving three dose administrations.