The current suite of Food and Drug Administration (FDA) approved antidotes (i.e., sodium nitrite, sodium thiosulfate, and hydroxocobalamin) are effective for treating cyanide poisoning, but individually, each antidote has major limitations (e.g., large effective dosage or delayed onset of action). To mitigate these limitations, next-generation cyanide antidotes are being investigated, including 3-mercaptopyruvate (3-MP) and cobinamide (Cbi). Analytical methods capable of detecting these therapeutics individually and simultaneously (for combination therapy) are essential for the development of 3-MP and Cbi as potential cyanide antidotes. Therefore, a liquid chromatography-tandem mass-spectrometry method for the simultaneous analysis of 3-MP and Cbi was developed. Sample preparation of 3-MP consisted of spiking plasma with an internal standard (13C3-3-MP), precipitation of plasma proteins, and derivatizing 3-MP with monobromobimane to produce 3-mercaptopyruvate-bimane. Preparation of Cbi involved denaturing plasma proteins with simultaneous addition of excess cyanide to convert each Cbi species to dicyanocobinamide (Cbi(CN)2). The limits of detection for 3-MP and Cbi were 0.5μM and 0.2μM, respectively. The linear ranges were 2-500μM for 3-MP and 0.5-50μM for Cbi. The accuracy and precision for 3-MP were 100±9% and <8.3% relative standard deviation (RSD), respectively. For Cbi(CN)2, the accuracy was 100±13% and the precision was <9.5% RSD. The method presented here was used to determine 3-MP and Cbi from treated animals and may ultimately facilitate FDA approval of these antidotes for treatment of cyanide poisoning.
|Original language||English (US)|
|Number of pages||8|
|Journal||Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences|
|State||Published - Jan 1 2016|
Bibliographical noteFunding Information:
We gratefully acknowledge the support from the CounterACT Program , National Institutes of Health Office of the Director (NIH OD) , and the National Institute of Allergy and Infectious Diseases , (Grant Number Y1-OD-0690-01/A-120-B.P2010-01 , Y1-OD-1561-01/A120-B.P2011-01 , AOD14020-001-00000/A120-B.P2014-01 , AOD13016-001-00000/A120-B.P2013-01 , AOD12060-001-00000/A120-B.P2012-01 ). We also would like to acknowledge support by U.S. Dept. of Education GAANN award ( P200A100103 ). We thank the National Science Foundation Major Research Instrumentation Program (Grant Number CHE-0922816 ) for funding the AB SCIEX QTRAP 5500 LC/MS/MS ( EPSCoR Grant 0091948 ). Lastly, we would like to thank the South Dakota State University Campus Mass Spectrometry Facility for the use of the LC–MS-MS, obtained with the support from the National Science Foundation / EPSCoR (Grant Number 0091948 ). The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the national Science Foundation, the National Institute of Health, or Department of Defense.
© 2015 Elsevier B.V.
- Cyanide antidote
- Mass spectrometry