Pharmaceutical Sciences Faculty Publications

Quantitative Determination of Antalarmin, a Novel Corticotropin-Releasing Hormone Receptor-1 Antagonist, in Canine Plasma by HPLC-MS

Document Type

Article

Publication Date

3-1-2005

Journal Title

Journal of Separation Science

ISSN

1615-9306

Volume

28

Issue

4

First Page

332

Last Page

336

DOI

10.1002/jssc.200400048

PubMed ID

15792247

Abstract

A simple and rapid method was developed for the quantitation of antalarmin from plasma using high performance liquid chromatography coupled with electrospray ionization mass spectrometry (ESI/MS). Separation of antalarmin from interfering compounds was achieved using reversed phase chromatography on a C-8 micro-column with an isocratic mobile phase comprised of 80% acetonitrile, 20% water, and 5 mM triethylamine. Detection by ESI/MS was accomplished in positive ion mode using single ion monitoring of the protonated molecular ions of antalarmin and its 13C2-isotopimer. The area ratio of the integrated peaks of interest in the extracted ion chromatogram was used for quantitation. The lower limit of detection was 1 picogram (pg) and the quantitation showed a linear response up to 4 nanograms loaded on column. To achieve acceptable accuracy at or around the limit of quantitation of 20 pg, a 1/x weighting was applied to the calibration data. Accuracy and precision variation for intra and inter-day validation were below the acceptable limit (15%) for pharmacokinetic studies.

Keywords

Chromatography, high pressure liquid, mass spectrometry, molecular structure, pyrimidines, pyrroles, receptors, corticotropin-releasing hormone

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