Quantitative analysis of the immunosuppressant CP-690,550 in whole blood by column-switching high-performance liquid chromatography and mass spectrometry detection

Ricardo Paniagua, Andrew Campbell, Paul S. Changelian, Bruce A. Reitz, Chandra Prakash, Dominic C. Borie

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

A fast and accurate method to quantify the new immunosuppressive JAK3 inhibitor CP-690,550 in whole blood using a dual-pump liquid chromatography-liquid chromatography-mass spectrometry (LC/LC-MS) system was developed and validated in nonhuman primate blood. Before injection, blood samples were prepared by precipitation with a reagent that included methanol and acetonitrile (30:70, vol/vol) along with the internal standard (CP-istd). Column-switching LC/LC-MS analysis used online extraction followed by separation on a C8 analytic column and MS detection of the [M + H]+ CP-690,550 (m/z = 313.1) and CP internal standard (m/z = 288.1). Linearity was always better than r2 = 0.99 (n = 7) for CP-690,550 (range 2.5-750 ng/mL), with a lower limit of quantification (LLOQ) of 2.5 ng/mL. The intrarun accuracy and precision ranged from 103.0% to 105.4% and 2.7% to 4.3%, respectively (n = 5), and the interday precision ranged from 8.7% to 11.1%, and the interday accuracy ranged from 98.1% to 103.8% of nominal values (n = 14). The injection repeatability for the method was 1.3% (n = 7). Except for the LLOQ, the intraday accuracy and precision in human blood were also within 15% (n = 5). The combination of simple sample preparation and short analytic run time of this sensitive procedure makes it effective for monitoring the concentration of CP-690,550 in whole blood in organ-transplant recipients.

Original languageEnglish (US)
Pages (from-to)608-616
Number of pages9
JournalTherapeutic Drug Monitoring
Volume27
Issue number5
DOIs
StatePublished - Oct 1 2005

Keywords

  • Immunosuppression
  • JAK/STAT
  • Liquid chromatography
  • Mass spectrometry
  • Transplantation

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)

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