A direct determination of thymidine triphosphate concentrations without dephosphorylation in peripheral blood mononuclear cells by LC/MS/MS.
A rapid, sensitive and specific analytical method with minimal sample preparation for the measurement of thymidine triphosphate (TTP) in peripheral blood mononuclear cells (PBMC) by LC/MS/MS has been developed. PBMC were separated from whole blood or buffy coat. The analyte and internal standard were extracted from PBMC with 70% methanol (pH 7.2). These extracts after centrifugation were directly injected onto LC/MS/MS without need for any further sample preparation. The calibration curve was linear over the range 0.8-800 ng/ml. Mean inter- and intra-assay coefficients of variation (CVs) over the range of the standard curve were less than 10%. The overall recovery of TTP was 103.5%.
Chi
J
J
Department of Clinical Pharmacy, Drug Research Unit, School of Pharmacy, University of California, San Francisco, CA 94143-0622, USA.
Jayewardene
A
A
Stone
J
J
Gambertoglio
J G
JG
Aweeka
F T
FT
D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D005615:Freezing; D006801:Humans; D066298:In Vitro Techniques; D009000:Monocytes / Q000737:chemistry*; D010766:Phosphorylation; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D013942:Thymine Nucleotides / Q000032:analysis*
D013942:Thymine Nucleotides; C024157:thymidine 5'-triphosphate