title: Development of a fast and simple liquid chromatography-tandem mass spectrometry method for the quantitation of argatroban in patient plasma samples.
abstract: An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the direct measurement of argatroban in human plasma was developed and compared with the activity-based Hemoclot Thrombin Inhibitors assay. UPLC-MS/MS was performed using diclofenac as an internal standard. In summary, argatroban and diclofenac were extracted from 100 μL of plasma using a methanol precipitation protocol, and chromatographic separation was performed on an ACQUITY TQD mass spectrometer using a UPLC C18 BEH 1.7 μm column with a water and methanol gradient containing 0.1% formic acid. The detection and quantitation were performed using positive ion electrospray ionization and multiple reaction monitoring (MRM) mode. The UPLC-MS/MS method was linear over the concentration range of 0.003-3.0 μg/mL, with a lower limit of quantitation for argatroban of 0.003 μg/mL. The intra- and inter-assay imprecision was less than 12% at the plasma argatroban concentrations tested. Good correlation was demonstrated between the UPLC-MS/MS method and the indirect activity-based assay for determination of argatroban. However, increased plasma fibrinogen levels caused underestimation of argatroban levels using the indirect activity-based assay, whereas the UPLC-MS/MS method was unaffected. UPLC-MS/MS provides a relatively simple, sensitive, and rapid means of argatroban monitoring. It has successfully been applied to assess plasma argatroban concentrations in hospitalized patients and may provide a more accurate determination of argatroban concentrations than an activity-based assay in certain clinical conditions.
authors.affiliation: Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA 30308, USA.
authors.forename: Jeanne M
authors.initials: JM
authors.lastname: Rhea
authors.forename: Marion L
authors.initials: ML
authors.lastname: Snyder
authors.forename: Anne M
authors.initials: AM
authors.lastname: Winkler
authors.forename: Charbel
authors.initials: C
authors.lastname: Abou-Diwan
authors.forename: Corinne R
authors.initials: CR
authors.lastname: Fantz
authors.forename: James C
authors.initials: JC
authors.lastname: Ritchie
authors.forename: Fania
authors.initials: F
authors.lastname: Szlam
authors.forename: Kenichi A
authors.initials: KA
authors.lastname: Tanaka
authors.forename: Ross J
authors.initials: RJ
authors.lastname: Molinaro
mesh_terms: D001120:Arginine / Q000031:analogs & derivatives; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016903:Drug Monitoring / Q000379:methods*; D006801:Humans; D016014:Linear Models; D010875:Pipecolic Acids / Q000097:blood*; D015203:Reproducibility of Results; D013449:Sulfonamides; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D010875:Pipecolic Acids; D013449:Sulfonamides; D001120:Arginine; C031942:argatroban