Simultaneous quantification of CTN986 and its deglycosylation products in rat serum using liquid chromatography/tandem mass spectrometry.
A quantitative method for the simultaneous determination of CTN986, a flavonol triglycoside, and its two deglycosylation products rutin and hirsutin in rat serum was developed and validated for the investigation of the pharmacokinetics of CTN986. Analytes were isolated from the serum samples (200 microL) prior to analysis by liquid chromatography/tandem mass spectrometry (LC/MS/MS) using C(18) solid-phase extraction, and were separated on a Zorbax C(8) reversed-phase column with an isocratic mobile phase consisting of methanol/isopropanol/water/formic acid (20:10:70:0.1, v/v/v/v). The protonated analytes generated in the positive ion mode were monitored through multiple reaction monitoring in an eletrospray ionization source. Calibration was performed by internal standardization with CTN987, a flavonoid structurally similar to CTN986, and regression curves were constructed ranging from 2 to 1000 ng/mL in 200 microL serum samples. The intra- and inter-day precision values were below 11% and accuracy was between -2.37 and 1.4% for all quality control samples. This quantitation method was successfully applied to pharmacokinetic studies of CTN986 in rats following oral and intravenous administration. Rutin and hirsutin were not detected in rat serum.