title: Application of automated serial blood sampling and dried blood spot technique with liquid chromatography-tandem mass spectrometry for pharmacokinetic studies in mice.
abstract: The goal of this work was to obtain full pharmacokinetic profiles from individual mice with the use of an automated blood sampling system and dried blood spot (DBS) technique. AMG 517, a potent and selective vanilloid receptor (VR1) antagonist, was dosed to mice (n=3) intravenously and blood samples were collected using the automated blood sampling system with the "no blood waste" method. The collected blood samples were a mixture of 25 μL blood and 50 μL of heparinized saline solution. Two 15 μL aliquots were manually spotted onto a DBS card and dried at room temperature for at least 2h before being stored in zip bags with desiccant. The remaining samples (45 μL) were stored at -70°C until analysis. Both the DBS and the whole blood samples (diluted with saline (1:2, v/v)) were extracted and analyzed by liquid chromatography-tandem mass spectrometry. The overall extraction recovery of the analyte from the dried blood spots was determined to be about 90%. The pharmacokinetic parameters calculated using the whole blood or the DBS concentration data were comparable, and were obtained from only 3 mice, whereas conventional sampling and analysis would have required up to 27 mice to achieve the same result. The analyte was shown to be stable in the diluted whole blood (blood:saline 1:2) at room temperature for at least 4h and in the DBS for at least 34 days when stored at room temperature. These results indicated that the automated blood sampling system and DBS collection are promising techniques to obtain full pharmacokinetic profiles from individual mice and reduce the use of animals.
authors.affiliation: Department of Pharmacokinetics and Drug Metabolism, Amgen, Thousand Oaks, CA 91320-1799, United States. pwong@amgen.com
authors.forename: Philip
authors.initials: P
authors.lastname: Wong
authors.forename: Roger
authors.initials: R
authors.lastname: Pham
authors.forename: Carl
authors.initials: C
authors.lastname: Whitely
authors.forename: Marcus
authors.initials: M
authors.lastname: Soto
authors.forename: Kevin
authors.initials: K
authors.lastname: Salyers
authors.forename: Christopher
authors.initials: C
authors.lastname: James
authors.forename: Bernd A
authors.initials: BA
authors.lastname: Bruenner
mesh_terms: D000818:Animals; D052160:Benzothiazoles / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D001800:Blood Specimen Collection / Q000379:methods*; D002138:Calibration; D002853:Chromatography, Liquid / Q000379:methods*; D019411:Clinical Laboratory Techniques / Q000379:methods; D059788:Dried Blood Spot Testing / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods; D008297:Male; D051379:Mice; D011743:Pyrimidines / Q000008:administration & dosage / Q000097:blood* / Q000493:pharmacokinetics; D012015:Reference Standards; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D052160:Benzothiazoles; C523409:N-(4-(6-(4-trifluoromethylphenyl)pyrimidin-4-yloxy)benzothiazol-2-yl)acetamide; D011743:Pyrimidines