title: 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.
abstract: 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.
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, No. 325 Guohe Road, Shanghai 200433, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xie
authors.forename: Hongbing
authors.initials: H
authors.lastname: He
authors.forename: Guorong
authors.initials: G
authors.lastname: Fan
authors.forename: Yutian
authors.initials: Y
authors.lastname: Wu
mesh_terms: D000818:Animals; D004285:Dogs; D015718:Fibrin Tissue Adhesive / Q000008:administration & dosage / Q000493:pharmacokinetics; D005340:Fibrinogen / Q000502:physiology*; D007274:Injections, Intraperitoneal; D057230:Limit of Detection; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D013552:Swine; D014014:Tissue Adhesives
chemical_list: D015718:Fibrin Tissue Adhesive; D014014:Tissue Adhesives; D005340:Fibrinogen