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High‐throughput determination of atrasentan in human plasma by high‐performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry
Author(s) -
Wang Perry G.,
Wei Jack,
Chang Min,
ElShourbagy Tawakol
Publication year - 2005
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.493
Subject(s) - chemistry , chromatography , mass spectrometry , detection limit , electrospray ionization , analytical chemistry (journal) , ammonium acetate , calibration curve , high performance liquid chromatography
Atrasentan (A‐147627) is an endothelin antagonist receptor being developed at Abbott Laboratories for the treatment of prostate cancer. A quick and sensitive method for the determination of atrasentan in human plasma has been developed and validated using high‐performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry. A dual‐column, single mass spectrometer system is used to provide a reliable and routine means to increase sample throughput. The analytical method involves liquid–liquid extraction and internal standard (A‐166790). The plasma samples and internal standard are acidified with 0.3 m hydrochloric acid prior to being extracted into 1:1 (v[sol ]v) hexanes–methyl t ‐butyl ether. The organic extract was evaporated to dryness using heated nitrogen stream and reconstituted with mobile phase. Atrasentan and internal standard were separated with no interference in a Zorbax SB‐C 18 analytical column with 2.1 × 50 mm, 5 µm, and a Zorbax C 8 guard column using a mobile phase consisting of 50:50 (v:v) acetonitrile–0.05 m ammonium acetate, pH 4.5, at a flow rate of 0.30 mL[sol ]min to provide 4 min chromatograms. For a 250 µL plasma sample volume, the limit of quantitation was approximately 0.3 ng[sol ]mL. The calibration was linear from 0.30 to 98.0 ng[sol ]mL ( r 2 > 0.995). A significant advantage of the method is the ability to employ parallel HPLC separations with detection by a single MS[sol ]MS system to provide sensitivity and selectivity sufficient to achieve robust analytical results with a lower limit of quantitation of 0.30 ng[sol ]mL and high throughput. Copyright © 2005 John Wiley & Sons, Ltd.