Sensitive and cost-effective LC-MS/MS method for quantitation of CVT-6883 in human urine using sodium dodecylbenzenesulfonate additive to eliminate adsorptive losses.
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.
Chen
Chungwen
C
Department of Pre-clinical Development, CV Therapeutics, Inc., 3172 Porter Drive, Palo Alto, CA 94304, USA. william.chen@cvt.com
Bajpai
Lakshmikant
L
Mollova
Nevena
N
Leung
Kwan
K
D000327:Adsorption; D001557:Benzenesulfonates / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000191:economics* / Q000379:methods*; D006801:Humans; D011687:Purines / Q000652:urine*; D011720:Pyrazoles / Q000652:urine*; D012015:Reference Standards; D053719:Tandem Mass Spectrometry / Q000191:economics* / Q000379:methods*
D001557:Benzenesulfonates; D011687:Purines; D011720:Pyrazoles; C001114:dodecylbenzenesulfonic acid; C518643:3-ethyl-1-propyl-8-(1-(3-trifluoromethylbenzyl)-1H-pyrazol-4-yl)-3,7-dihydropurine-2,6-dione