Measurement of low picomolar levels of triamcinolone acetonide in human bronchoalveolar lavage fluid by gas chromatography-electron-capture negative-ion mass spectrometry

Walter C. Hubbard, Mark C. Liu, Carol Bickel, Domenick Argenti, Don Heald, Robert P. Schleimer

Research output: Contribution to journalArticle

Abstract

The intense inherent electron-capture properties of the C21 acetate derivative of triamcinolone acetonide (TAA) under methane chemical ionization mass spectrometric conditions were exploited for the development of a highly sensitive and selective gas chromatography-mass spectrometric (GC-MS) technique for measurement of levels of TAA in human bronchoalveolar lavage (BAL) fluid. After the addition of 3.0 ng of a heptadeuterated analog of TAA and varying concentrations of TAA to 2-ml aliquots of BAL fluid, the deuterium and protium forms of the steroid were extracted with diethyl ether, converted to the C21 acetate derivative, and purified via adsorptive chromatography prior to GC-MS analysis. Standard curves obtained from 2-ml aliquots of BAL fluid were linear over a wide range of concentrations of TAA from 0.0 to 24,600 pg/2-ml aliquots of BAL fluid. Levels as low as 6.0 pg/ml (13.8 pmol L-1) in BAL fluid can be reliably determined in 2-ml aliquots of the biological fluid with <10% error. These findings suggest that the assay method exploiting the intense electron-capture properties of TAA is highly suitable for determination of the deposition pattern and in vivo kinetics of TAA in human airways following inhalation of the steroid.

Original languageEnglish (US)
Pages (from-to)18-25
Number of pages8
JournalAnalytical biochemistry
Volume290
Issue number1
DOIs
StatePublished - Mar 1 2001

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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