Determination of the transplacental transfer of paclitaxel and antipyrine by high performance liquid chromatography coupled with photodiode array detector
Ex vivo placental perfusion experiments are important in understanding the quantity and mechanisms of xenobiotic transport to the fetus during pregnancy. Our study demonstrates that paclitaxel and antipyrine concentrations in placental perfusion medium containing physiological concentrations of human serum albumin during pregnancy (30 mg/mL) can be quantified by RP-HPLC and UV detection. A liquid-liquid extraction method was developed for the quantification of paclitaxel and celecoxib (internal standard) from perfusion medium. Antipyrine, which is a necessary marker in placental perfusions for determining the validity of experiments and calculating the clearance index of xenobiotics, was also analyzed by HPLC and UV detection. Antipyrine concentrations were determined by HPLC after precipitating the perfusion medium in acetonitrile and separating the precipitated proteins by centrifugation. Concentrations were fitted to linear regressions with R2 values approaching 1. Lower limits of detection for paclitaxel and antipyrine were 100 ng/mL and 200 ng/mL, respectively. Both methods demonstrated high intra-day and inter-day precision and trueness. Additionally, the use of these methods was demonstrated in a placental perfusion experiment using Taxol® (paclitaxel dissolved in Cremophor-EL). The fetal transfer rate of Taxol was 6.6% after 1 hour.