Imatinib assay by HPLC with photodiode-array UV detection in plasma from patients with chronic myeloid leukemia: Comparison with LC-MS/MS

Clin Chim Acta. 2010 Feb;411(3-4):140-6. doi: 10.1016/j.cca.2009.10.007. Epub 2009 Oct 22.

Abstract

Background: Imatinib, a competitive inhibitor of BCR-ABL tyrosine kinase, is now the first-line treatment for chronic myelogenous leukemia (CML). Therapeutic drug monitoring targeting trough plasma levels of about 1000ng/mL may help to optimize the therapeutic effect.

Methods: We developed a high-performance liquid chromatography (HPLC) method with UV/Diode Array Detection (DAD) for trough imatinib concentration determination in human plasma. Imatinib trough levels were measured in plasma from 65 CML patients using our method and LC-MS/MS as the reference method. Results with these two methods were compared using Deming regression, chi-square test, and sign test.

Results: The calibration curve was prepared in blank human plasma. HPLC-UV/DAD calibration curves were linear from 80 to 4000ng/mL, and the limit of quantification was set at 80ng/mL. The between-day variation was 6.1% with greater than 96% recovery after direct plasma deproteinization and greater than 98% recovery from the column. No significant differences in imatinib plasma levels were found between HPLC-UV/DAD and LC-MS/MS.

Conclusions: This HPLC-UV/DAD method was sufficiently specific and sensitive for imatinib TDM, with no evidence of interference. Our rapid inexpensive HPLC-UV/DAD method that requires only widely available equipment performs well for plasma imatinib assays.

Publication types

  • Comparative Study

MeSH terms

  • Benzamides
  • Blood Chemical Analysis / economics
  • Blood Chemical Analysis / instrumentation
  • Blood Chemical Analysis / methods*
  • Blood Proteins / isolation & purification
  • Chromatography, High Pressure Liquid
  • Electric Conductivity
  • Female
  • Humans
  • Imatinib Mesylate
  • Laboratories, Hospital
  • Leukemia, Myelogenous, Chronic, BCR-ABL Positive / blood*
  • Limit of Detection
  • Linear Models
  • Male
  • Middle Aged
  • Piperazines / blood*
  • Piperazines / isolation & purification
  • Pyrimidines / blood*
  • Pyrimidines / isolation & purification
  • Reproducibility of Results
  • Tandem Mass Spectrometry
  • Temperature
  • Time Factors
  • Ultraviolet Rays*

Substances

  • Benzamides
  • Blood Proteins
  • Piperazines
  • Pyrimidines
  • Imatinib Mesylate