Development of a High-Sensitivity Quantitation Method for Arginine Vasopressin by High-Performance Liquid Chromatography Tandem Mass Spectrometry, and Comparison with Quantitative Values by Radioimmunoassay.

Abstract

Human plasma arginine vasopressin (AVP) levels serve as a clinically relevant marker of diabetes and related syndromes. We developed a highly sensitive method for measuring human plasma AVP using high-performance liquid chromatography tandem mass spectrometry. AVP was extracted from human plasma using a weak-cation solid-phase extraction plate, and separated on a wide-bore octadecyl reverse-phase column. AVP was quantified in ion-transition experiments utilizing a product ion (m/z 328.3) derived from its parent ion (m/z 542.8). The sensitivity was enhanced using 0.02% dichloromethane as a mobile-phase additive. The lower limit of quantitation was 0.200 pmol/L. The extraction recovery ranged from 70.2 ± 7.2 to 73.3 ± 6.2% (mean ± SD), and the matrix effect ranged from 1.1 - 1.9%. Quality-testing samples revealed interday/intraday accuracy and precision ranging over 0.9 - 3% and -0.3 - 2%, respectively, which included the endogenous baseline. Our results correlated well with radioimmunoassay results using 22 human volunteer plasma samples.