title: Evaluation of dried blood spot (DBS) technology versus plasma analysis for the determination of MK-1775 by HILIC-MS/MS in support of clinical studies.
abstract: The collection of human blood samples as dried blood spots (DBS) for the pharmacokinetic assessment of investigational drugs in clinical trials offers a number of advantages over conventional plasma sampling, namely, small sample volume, simplified sample handling, and cost-effective shipping and storage. The use of DBS coupled with liquid chromatography-tandem mass spectrometry analysis was evaluated for the quantification of MK-1775, a Wee-1 inhibitor under development as a chemo/radio-sensitizer for the treatment of cancer. The DBS method exhibited an assay performance comparable to that of the existing plasma assay, which is currently used in support of clinical studies. Both assays used the same linear dynamic range of 2-1,000 ng/mL, with a lower limit of quantification of 2 ng/mL. Based on the intra-day assay validation results, the accuracy of the DBS method ranged from 94.0 to 105.0%, with a coefficient of variation of <4.8%. The blood-to-plasma ratio calculated from the DBS data (blood concentrations) and the plasma data (plasma concentrations) was in good agreement with the one obtained from the in vitro assessment using conventional methodology. No significant hematocrit impact on the assay was observed as hematocrit ranged from 16 to 85%. The correlation between the measured MK-1775 concentrations in plasma and that determined in dried blood spots from oncology patients during the ongoing clinical study was discussed.
authors.affiliation: Department of Pharmacokinetics, Pharmacodynamics and Drug Metabolism, Regulated Bioanalysis, Merck Research Laboratories, WP75B-300, West Point, PA 19486, USA. yangxu15@hotmail.com
authors.forename: Yang
authors.initials: Y
authors.lastname: Xu
authors.forename: Wei
authors.initials: W
authors.lastname: Fang
authors.forename: Wei
authors.initials: W
authors.lastname: Zeng
authors.forename: Suzanne
authors.initials: S
authors.lastname: Leijen
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Woolf
mesh_terms: D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D059788:Dried Blood Spot Testing / Q000191:economics / Q000379:methods*; D016903:Drug Monitoring / Q000191:economics / Q000379:methods*; D006400:Hematocrit; D006801:Humans; D057230:Limit of Detection; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D011720:Pyrazoles / Q000097:blood*; D011743:Pyrimidines / Q000097:blood*; D011744:Pyrimidinones; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D011720:Pyrazoles; D011743:Pyrimidines; D011744:Pyrimidinones; D011505:Protein-Tyrosine Kinases; C499349:WEE1 protein, human; C549567:adavosertib