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Journal of Pharmacology And Experimental Therapeutics, Vol. 112, Issue 1, 116-126, 1954
Copyright © 1954 by American Society for Pharmacology and Experimental Therapeutics

THE CHEMICAL MEASUREMENT OF HISTAMINE IN BLOOD PLASMA AND CELLS

Oliver H. Lowry 1, Helen Tredway Graham 1, Frances B. Harris 1, Martha K. Priebat 1, Ansel R. Marks 1, and Robert U. Bregman 1

1 Department of Pharmacology, Washington University School of Medicine, St. Louis, Missouri

1. A microchemical method permitting rapid and accurate determination of histamine in amounts as small as 0.01 microgm. and concentrations as low as 4 x 10-10 is described. The method is based on spectrophotometric observation of the optical density at lgr360 mµ of the dinitrofluorobenzene derivative of histamine. Sensitivity is secured through concentration of weak histamine solutions as much as 400 fold, in part by means of adsorption on Decalso columns, and in part by the extraction of the yellow derivative into methyl-n-hexyl ketone and thence into strong hydrochloric acid. Simultaneously, interfering substances are largely eliminated.

2. With this method the average amount of histamine found in human plasma is 4.3 microgm. per liter, of which only 60 per cent is destroyed by histaminase. Therefore the true plasma histamine is less than 3 microgm. per liter.

3. Histamine is 2,000 times as concentrated in the buffy coat (8,000 microgm. per liter) as in plasma. Most of this chromogenic material may be destroyed by histaminase. The buffy coat contains over 90 per cent of the whole blood histamine.

4. Erythrocytes contain relatively large amounts of interfering substances, probably including spermidine and spermine, which are not attacked by hisaminase. Therefore, the proposed method is not suitable for analysis of red cells, which in any event seem to contain quantitatively insignificant amounts of histamine.

Submitted on May 24, 1954




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Copyright © 1954 by the American Society for Pharmacology and Experimental Therapeutics.