Detection and quantitation of fatty acid acyl conjugates of triamcinolone acetonide via gas chromatography-electron-capture negative-ion mass spectrometry.
The inherent electron-capture properties of triamcinolone acetonide (TAA) fatty acid conjugates were exploited for development of a GC-MS technique for quantitation of C21 long-chain fatty esters of TAA synthesized in BEAS-2B cells, an immortalized airway epithelium cell line. TAA esters extracted from BEAS-2B cells were purified and detected via selected ion monitoring of the molecular anions generated from the TAA esters under electron-capture negative-ion mass spectrometric conditions. Standard curves were linear over a range of 0.0 to >4.5 ng/mg protein with r(2) values = 1. Levels of TAA conjugates extracted from BEAS-2B treated with 10(-5)M TAA for 24h ranged from 0.024 to 0.301 ng/mg protein. Further evidence for confirmation of the identity of TAA fatty esters formed in BEAS-2B cells was obtained via selected reaction monitoring. The transition monitored was formation of the carboxy anion generated from each of the respective molecular anions of the TAA esters during collision-induced decomposition. These findings indicate that the GC-MS analysis is suitable for studies of the kinetics of the TAA fatty acid conjugates formation in vitro and may be directly applicable to determination of the kinetics of TAA fatty acid conjugation in vivo.
Division of Allergy and Clinical Immunology, The Johns Hopkins University School of Medicine, Baltimore, MD 21224, USA. whubbard@jhmi.edu
Walter C
WC
Hubbard
Andrew E
AE
Blum
Carol A
CA
Bickel
Nicola M
NM
Heller
Robert P
RP
Schleimer
D000890:Anti-Infective Agents / Q000032:analysis* / Q000378:metabolism; D001980:Bronchi / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D004583:Electrons; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods* / Q000592:standards; D006801:Humans; D012680:Sensitivity and Specificity; D014221:Triamcinolone / Q000032:analysis* / Q000378:metabolism
D000890:Anti-Infective Agents; D005227:Fatty Acids; D014221:Triamcinolone