A simple, rapid and sensitive semimicro method for the measurement of cyanide in blood

Conventional methods do not meet the clinical need for rapid cyanide measurements. We report a procedure which can provide a result in 10 min. It should be of particular interest to laboratories serving cardiac or renal units which use the hypotensive agent sodium nitroprusside, and burns units to which fire victims may be admitted suffering the effects of HCN from inhaled smoke. A sample of blood (100 microL) is mixed with H3PO4, containing a surfactant, and the HCN is trapped in an alkaline mixture of 1,2-dinitrobenzene and 4-nitrobenzaldehyde in 2-methoxyethanol. The catalytic action of cyanide, which produces purple 2-nitrophenylhydroxylamine, is stopped with acetone after 6 min. The absorbance measured at 560 nm shows a linear relationship with the cyanide concentration, but the slope varies with the ambient temperature. Since KCN added to both 50 mmol/L NaOH and blood gives similar assay results, any inaccuracy arising from changes in temperature can be avoided by running standards at the same time as the blood sample.