Comparison of Spectrophotometric Methods for the Determination of Carboxyhemoglobin in Postmortem Blood

Carbon monoxide (CO) poisoning presents an interesting challenge for postmortem toxicology laboratories. The discontinuation of the CO-oximeter manufactured by Instrumentation Laboratories has left many forensic laboratories without this simple but reliable choice for the analysis of CO in blood. A comparable alternative that is quick and simple is analysis using a standard ultraviolet-visible spectrophotometer, which offers sufficient precision and accuracy for the measurement of percentage carboxyhemoglobin (%COHb) in postmortem blood. The hurdle for laboratories then becomes the selection of an appropriate spectrophotometric method since a variety of procedures have been published over the years. Four methods were evaluated based on literature findings and/or the appropriateness for postmortem testing. These methods are based on (i) relating %COHb to the ratio of COHb and Hb absorbance of a two-component system (two wavelengths), (ii) the multicomponent analysis of all hemoglobin species, (iii) the multicomponent analysis of a two-component system, and (iv) derivative spectroscopy. While all four methods performed similarly in terms of typical validation requirements, the ability to more effectively handle decomposed samples and the ease of sample preparation afforded (v) the multicomponent analysis of a two-component system-the most suitable for routine postmortem testing.