Acids from whole lipopolysaccharide and free lipid A of the closely related bacteria Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus were determined by gas chromatography and gas chromatography-mass spectrometry. In whole lipopolysaccharide, 3-hydroxymyristic acid was most abundant, followed by myristic and 3-deoxy-D-manno-2-octulosonic acid. In the lipid A moiety, myristic acid dominated, followed by 3-hydroxymyristic acid. The acid composition of whole lipopolysaccharide and free lipid A from A. actinomycetemcomitans and H. aphrophilus was not so specific as to allow taxonomic differentiation between these bacteria. If fatty acids of lipopolysaccharide are essential for expression of endotoxicity, the present results suggested no marked difference in the endotoxic activities of A. actinomycetemcomitans and H. aphrophilus.