An analytical method utilizing electrospray ionization ultra high-performance liquid chromatography tandem mass-spectrometry was developed for the quantification of cyanide metabolite a-ketolgutarate cyanohydrin (a-KgCN) in swine plasma. Swine plasma was spiked with a-KgCN, a-KgCN-d4 (internal standard), acidified with 1% formic acid in acetonitrile and centrifuged to precipitate plasma proteins.  After centrifugation, the supernatant was dried under N2 (g) and reconstituted in 1% formic acid.  Finally, samples were syringe-filtered and separated on a Phenomenex Synergi 2.5� Fusion-RP 100� (50 x 2.00 mm) column with gradient elution.  The method produced a dynamic range of 0.3-50 �M and a detection limit of 0.2 �M for a-KgCN.  Furthermore, the method produced a coefficient of variation of less than 13% with accuracies within 10% of the nominal concentrations for all intra- and inter-assay analyses.  The method was applied to the analysis of swine plasma following cyanide exposure.

Quantification of cyanide metabolite α-ketoglutarate cyanohydrin in plasma by ultra high-performance liquid chromatography tandem mass spectrometry

245th National Meeting (2013)

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