A Rapid Spectrophotometric 5me’l’i101> for the Determination of Methemoglobin and Carbonylhemoglobin in Blood

Little use has thus far been made of the near infra-red spectrum for the identification and determination of hemoglobin and its derivatives BJ means of infra-red photography Eggert (1) established the transparency of carbon monoxide-saturated blood and used this property for the qualitative detection of carbon monoxide poisoning.. Eggert’s ‘observations were extended and placed on a more quantitative basis by Mcrkelbach (2). A furt,hcr advance n-as made by Matthes and Gross (3) who developed a method for the determination of the carbon monoxide content of blood i71 I,&O in which the absorption of: red and infra-red radiation by flowing blood was measured in the ear lobe. A band for reduced hemoglobin at 7550 A has been described by Sidwell, Munch, Barron, and Hogness (4) and employed by these Tvorkers for the spectrophotometric determination of the degree of oxygenation of hemoglobin solut,ions. In the near infra-red region of the spectrum, between 7000 and 10,000 A, the hemoglobin derivatives have absorption bands which, because of their lower specific absorption, become apparent only when solutions more concentrated than those generally employed for spectrophotometric work are examined. A detailed description of these bands is contained in a previous publication (5). Despite the lower absorption, the large differences between the spectra of methemoglobin and cyanmethemoglobin and between oxyhemoglobin and carbonylhemoglobin mako this region wll suited for analytical purposes. On the basis of these considerations \ve hart developed a spectrophotometric method for the determination of methemoglobin and carbonylhemoglobin in blood. The method is rapid and accurate and requires only a few manipulat,ions. It has an advantage over existing methods in that it permits t,he determination of the total hemoglobin, carbonylhemoglobin, and methemoglobin content in a single blood sample. Principle of M&od