Determination of porcine fibrinogen in rat and dog plasma after intraperitoneal injection of a porcine-derived fibrin glue by fluorescein-labeled assay method: Comparison with isotope-labeled assay method.

A sensitive and specific fluorescein isothiocyanate (FITC) label coupled with size-exclusion high-performance liquid chromatography-fluorescence detection (SE-HPLC-FLD) method was developed and validated for the estimation of the pharmacokinetic profiles of porcine fibrinogen after intraperitoneal injection of a porcine-derived fibrin glue (FG) to SD rats and beagle dogs with three single doses. Porcine fibrinogen, the major composition of the FG, was labeled with FITC. The FG containing FITC-labeled porcine fibrinogen was intraperitoneally administered to SD rats at three single dosages (100, 200, 400mg/kg of porcine fibrinogen), and the collected plasma was then detected by SE-HPLC-FLD method. The present technique was compared to the previously introduced isotope-labeled assay method for the pharmacokinetic studies in SD rats. The pharmacokinetic studies in SD rats showed that the correlation coefficient between the FITC-labeled assay and (125)I-labeled assay methods was r(2)=0.989. Thus, this FITC-labeled assay method performed well and demonstrated high concordance with the previous (125)I-labeled assay method, suggesting that FITC-labeled assay could substitute the (125)I-labeled assay as a method of choice for quantification in beagle dogs. Then the plasma levels of porcine fibrinogen in beagle dogs were studied by the FITC-labeled assay method with three single doses (15, 30, 60mg/kg of porcine fibrinogen). The method validation showed that the FITC label coupled with SE-HPLC-FLD method was suitable for the quantification of porcine fibrinogen in plasma samples with satisfactory linear (r(2)>0.999), precision (<12%), accuracy (95.5-104.9%) and recovery (>88%). The results showed linear disposition of porcine fibrinogen at the examined dosage range in SD rats or beagle dogs.