Chromatography of Tissue Histamine

Abstract

EXTRACTS of tissues exceptionally rich in histamine, such as ox pleura, ox liver capsule, and mast cell tumours from dogs¹, were prepared for ascending chromatographic estimation of histamine by simple extraction or by grinding the tissues with 10 per cent aqueous trichloracetic acid and centrifuging. Using as solvent mixture n-butanol: acetic acid: water (4 : 1 : 5), we observed that after spraying with the Pauly or the paranitraniline diazo-reagent two sharply defined spots appeared on the chromatogram of R _F = 0.11 and 0.65 (Fig. 1,A). Both spots contained material which when eluted with water or 0.01 N hydrochloric acid showed histamine activity on the isolated guinea pig ileum and on the blood pressure of the atropinized cat. Prior removal of the excess trichloracetic acid from the solution by ether extraction merely reduced the intensity of the fast-running spot. When an eluate of this fast-running spot (R _F = 0.65) was re-run in the same solvent as before, the histamine now remained entirely in its lower location (R _F = 0.11).