Quantitative determination of busulfan in human plasma by UPLC

Quanyun Alan Xu, Reza Kazerooni, Jay K. Thapar, Borje D. Andersson, Timothy L. Madden

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

Abstract

A rapid and reliable UPLC method was developed and validated for the determination of busulfan in human plasma. After protein precipitation, derivatization, and liquid-liquid extraction, separation of derivatized busulfan was achieved on an Acquity BEH C18 column using a gradient mobile phase consisting of a trifluoroacetic acid aqueous solution (0.2%, v/v) and acetonitrile. The column temperature was maintained at 50 °C and UV detection was carried out at 254 nm. The complete analytical run time was 1.3 min, 7-fold faster than our previous LC methodology. Quantification was performed using external standardization and calibration curves were linear (r ≥ 0.999) over the dynamic range of 0.05-5.00 μg mL-1. Intra-day and inter-day coefficients of variation were ≤6.9 and 3.9%, respectively, across the range of concentrations. Accuracy of the analytical method expressed as the relative error percentage was better than 5.4%. LOD and LOQ were 0.013 and 0.025 μg mL-1, respectively. Data obtained using the UPLC method was compared to those obtained from our previously used LC method by Deming regression analysis. The UPLC method was accurate, sensitive, and greatly increased sample analysis throughput as compared to our previous LC methodology allowing for a 4-fold increase in the number of patients who could be monitored during transplant therapy.

Original languageEnglish (US)
Pages (from-to)1505-1510
Number of pages6
JournalChromatographia
Volume70
Issue number9-10
DOIs
StatePublished - Nov 2009

Keywords

  • Bone marrow transplantation
  • Busulfan
  • Human plasma
  • Ultra performance liquid chromatography

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry
  • Organic Chemistry

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