Determination of 3-mercaptopyruvate in rabbit plasma by high performance liquid chromatography tandem mass spectrometry.
Accidental or intentional cyanide poisoning is a serious health risk. The current suite of FDA approved antidotes, including hydroxocobalamin, sodium nitrite, and sodium thiosulfate is effective, but each antidote has specific major limitations, such as large effective dosage or delayed onset of action. Therefore, next generation cyanide antidotes are being investigated to mitigate these limitations. One such antidote, 3-mercaptopyruvate (3-MP), detoxifies cyanide by acting as a sulfur donor to convert cyanide into thiocyanate, a relatively nontoxic cyanide metabolite. An analytical method capable of detecting 3-MP in biological fluids is essential for the development of 3-MP as a potential antidote. Therefore, a high performance liquid chromatography tandem mass spectrometry (HPLC-MS-MS) method was established to analyze 3-MP from rabbit plasma. Sample preparation consisted of spiking the plasma with an internal standard ((13)C3-3-MP), precipitation of plasma proteins, and reaction with monobromobimane to inhibit the characteristic dimerization of 3-MP. The method produced a limit of detection of 0.1μM, a linear dynamic range of 0.5-100μM, along with excellent linearity (R(2)≥0.999), accuracy (±9% of the nominal concentration) and precision (<7% relative standard deviation). The optimized HPLC-MS-MS method was capable of detecting 3-MP in rabbits that were administered sulfanegen, a prodrug of 3-MP, following cyanide exposure. Considering the excellent performance of this method, it will be utilized for further investigations of this promising cyanide antidote.
Department of Chemistry and Biochemistry, South Dakota State University, Avera Health and Science Center 131, Box 2202, Brookings, SD, 57007, USA.
Michael W
MW
Stutelberg
Department of Chemistry and Biochemistry, South Dakota State University, Avera Health and Science Center 131, Box 2202, Brookings, SD, 57007, USA.
Chakravarthy V
CV
Vinnakota
Department of Chemistry and Biochemistry, South Dakota State University, Avera Health and Science Center 131, Box 2202, Brookings, SD, 57007, USA.
Brendan L
BL
Mitchell
Center for Drug Design, University of Minnesota, 516 Delaware Street SE, Minneapolis 55455, MN, USA.
Alexandre R
AR
Monteil
Center for Drug Design, University of Minnesota, 516 Delaware Street SE, Minneapolis 55455, MN, USA.
Steven E
SE
Patterson
Department of Chemistry and Biochemistry, South Dakota State University, Avera Health and Science Center 131, Box 2202, Brookings, SD, 57007, USA. Electronic address: brian.logue@sdstate.edu.
Brian A
BA
Logue
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003545:Cysteine / Q000031:analogs & derivatives* / Q000097:blood / Q000737:chemistry; D004355:Drug Stability; D057230:Limit of Detection; D016014:Linear Models; D011817:Rabbits; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods
C008366:3-mercaptopyruvic acid; D003545:Cysteine
3-Mercaptopyruvate; Cyanide antidote; Liquid chromatography-tandem mass spectrometry; Sulfanegen