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Clinical Chemistry 29: 2061-2067, 1983;
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Clinical Chemistry, Vol 29, 2061-2067, Copyright © 1983 by American Association for Clinical Chemistry

The "HemoQuant" test: a specific and quantitative determination of heme (hemoglobin) in feces and other materials

S Schwartz, J Dahl, M Ellefson and D Ahlquist

We describe a new, specific, quantitative method for fecal blood, based on conversion of nonfluorescing heme to fluorescing porphyrins, that obviates serious deficiencies inherent in currently used tests. A two- reagent system is used to determine the two hemoglobin-related fractions that are found in feces. The hot citric acid extract includes only the variable fraction of porphyrins that have been preformed from heme in the intestinal tract; this often is the major fraction. Total hemoglobin is indirectly determined by reaction with heated oxalic acid:FeSO4 reagent, which converts the remaining heme to porphyrin without loss of the preformed porphyrins. A three-step purification procedure eliminates interfering materials. Analytical recovery of added hemoglobin is linearly related to concentration over a several- thousand-fold range. The assay is equally applicable to whole blood or to sub-microgram amounts of hemoglobin in the 8-mg (wet weight) fecal sample tested. Quality control by liquid chromatographic and fluorometric analysis documents fluorescence specificity of the heme- derived porphyrins.


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Copyright © 1983 by the American Association for Clinical Chemistry.