Sensitive and cost-effective LC-MS/MS method for quantitation of CVT-6883 in human urine using sodium dodecylbenzenesulfonate additive to eliminate adsorptive losses

J Chromatogr B Analyt Technol Biomed Life Sci. 2009 Apr 1;877(10):943-7. doi: 10.1016/j.jchromb.2009.02.045. Epub 2009 Feb 24.

Abstract

CVT-6883, a novel selective A(2B) adenosine receptor antagonist currently under clinical development, is highly lipophilic and exhibits high affinity for non-specific binding to container surfaces, resulting in very low recovery in urine assays. Our study showed the use of sodium dodecylbenzenesulfonate (SDBS), a low-cost additive, eliminated non-specific binding problems in the analysis of CVT-6883 in human urine without compromising sensitivity. A new sensitive and selective LC-MS/MS method for quantitation of CVT-6883 in the range of 0.200-80.0ng/mL using SDBS additive was therefore developed and validated for the analysis of human urine samples. The recoveries during sample collection, handling and extraction for the analyte and internal standard (d(5)-CVT-6883) were higher than 87%. CVT-6883 was found stable under the following conditions: in extract - at ambient temperature for 3 days, under refrigeration (5 degrees C) for 6 days; in human urine (containing 4mM SDBS) - after three freeze/thaw cycles, at ambient temperature for 26h, under refrigeration (5 degrees C) for 94h, and in a freezer set to -20 degrees C for at least 2 months. The results demonstrated that the validated method is sufficiently sensitive, specific, and cost-effective for the analysis of CVT-6883 in human urine and will provide a powerful tool to support the clinical programs for CVT-6883.

Publication types

  • Evaluation Study

MeSH terms

  • Adsorption
  • Benzenesulfonates / chemistry*
  • Chromatography, High Pressure Liquid / economics*
  • Chromatography, High Pressure Liquid / methods*
  • Humans
  • Purines / urine*
  • Pyrazoles / urine*
  • Reference Standards
  • Tandem Mass Spectrometry / economics*
  • Tandem Mass Spectrometry / methods*

Substances

  • Benzenesulfonates
  • Purines
  • Pyrazoles
  • dodecylbenzenesulfonic acid
  • 3-ethyl-1-propyl-8-(1-(3-trifluoromethylbenzyl)-1H-pyrazol-4-yl)-3,7-dihydropurine-2,6-dione