Summary
The possible chemical mechanisms by which neutral solutions of sodium hypochlorite containing a high concentration of sodium chloride abolish the acidophilia of proteins in sections of fixed tissue are reviewed. The most probable one is the chlorination of the protein terminal amino groups, followed by the breakdown of the N-chloramine so formed into α-ketocarboxylic acid, nitrile or aldehyde groups. Hypochlorite solutions certainly do not deaminate tissue sections as was previously thought.
Experimental evidence for the formation of relatively stable N-chloramine groups in situ and their limited conversion to aldehydes is reported. For example, the acidophilia of hypochlorite-treated sections was found to be restored after flooding them with hydriodic acid followed by the extraction of the liberated iodine with an alcohol. The significance of these experimental findings is discussed.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Alfert, M., Geschwind, I. I.: A selective staining method for basic proteins of cell nuclei. Proc. nat. Acad. Sci. (Wash.) 39, 991–999 (1953)
Arx, E. von, Neher, R.: Eine multidimensionale Technik zur chromatographischen Identifizierung von Aminosäuren. J. Chromatog. 12, 329–341 (1963)
Aubel, E., Asselineau, J.: Recherches sur l'emploi de l'hypochlorite de sodium dans le dosage de l'alanine, la valine et leucines. Biochim. biophys. Acta (Amst.) 2, 198–206 (1948)
Baker, R. W. R.: Studies on the reaction between sodium hypochlorite and proteins. 1. Physico-chemical study of the course of the reaction. Biochem. J. 41, 337–342 (1947)
Burstone, M. S.: An evaluation of histochemical methods for protein groups. J. Histochem. Cytochem. 3, 32–49 (1955)
Cahn, J. W., Powell, R. E.: The Raschig synthesis of hydrazine. J. Amer. chem. Soc. 76, 2565–2567 (1954)
Chu, C. H. U., Fogelson, M. H., Swinyard, C. A.: A histochemical test for alpha amino acids. J. Histochem. Cytochem. 1, 391 (1953)
Dakin, H. D.: Oxidation of amino acids to cyanides. Biochem. J. 10, 319–323 (1916)
Dakin, H. D.: On the oxidation of amino acids and of related substances with chloramine-T. Biochem. J. 11, 79–95 (1917)
Dakin, H. D., Cohen, J. B., Daufrensne, M., Kenyon, J.: The antiseptic action of substances of the chloramine group. Proc. roy. Soc. B 89, 232–251 (1916)
Feigl, F., Rosell, R. A.: Nachweis von organischen Oxydantien in der Tüpfelanalyse. Unterscheidung von Chloramin T und Alkalihypochlorit. Analyt. Chem. 159, 335–339 (1957)
Goldschmidt, S., Steigerwald, C.: Über den Abbau von Proteinen durch Hypobromit. Ber. 58, 1346–1353 (1925)
Herken, H., Silbersiepe, H. O.: Zur Spaltung von Peptidbindungen in Proteinen durch Hypochlorit. Arch. exp. Path. 212, 205–213 (1951)
Ingols, R. S., Wyckoff, H. A., Kethley, T. W., Hodgden, H. W., Fincher, E. L., Hildebrand, J. C., Mandel, J. E.: Bacterial studies of chlorine. Ind. Eng. Chem. 45, 996–1000 (1953)
Jaksic, M. M., Nikolic, B. Z., Csonka, I. M., Dordevic, A. B., Karanovic, D. M.: Neutral salts effect on available chlorine conversion to chlorate. Chem. Abs. 71, abstract no. 42746 (Abstract of russian article in Kem. Ind. 17, 707–711) (1969)
Langheld, K.: Über das Verhalten von α-Aminosäuren gegen Natriumhypochlorit. Ber. 42, 2360–2375 (1909)
Pereira, W. E., Hoyano, Y., Summons, R. E., Bacon, V. A., Duffield, A. M.: Chlorination studies II. The reaction of aqueous hypochlorous acid with α-amino acids and dipeptides. Biochim. biophys. Acta (Amst.) 313, 170–180 (1973)
Raschig, F.: Vorlesungversuche aus der Chemie der anorganischen Stickstoffverbindungen. Ber. 40, 4580–4588 (1907)
Robson, H. L.: Chloramines and chloroamines. In Kirk-Othmer's Encyclopedia of Chemical Technology. 2nd ed., vol. 4, p. 908–928. New York: Interscience 1964
Ruddell, C. L.: The demonstration of anions generated by the action of sodium hypochlorite (NaOCl) on tissue sections, including observations on the unmasking of carbonyl groups. Histochemie 19, 319–339 (1969)
Rydon, H. N., Smith, P. W. G.: A new method for the detection of peptides and similar compounds on paper chromatograms. Nature (Lond.) 169, 922–923 (1952)
Smith, L.: Die Bildungsweise der Chlorhydrine. I. Mitteilung: Glyzerinchlorhydrine. Z. physikal. Chem. 92, 717–740 (1918)
Stoward, P. J.: Histochemical studies of the formazan reaction, II. The conversation of periodate-reactive mucosubstances into diphenyl- and phenyl-4′-diazo-3,3′-dimethoxybiphenyl formazans and related derivatives. J. roy. micr. Soc. 87, 407–435 (1967 a)
Stoward, P. J.: Studies in fluorescence histochemistry, III. The demonstration with salicylhydrazide of the aldehydes present in periodate-oxidized mucosubstances. J. roy. micr. Soc. 87, 247–257 (1967 b)
Stoward, P. J.: The histochemical properties of some periodate-reactive mucosubstances of the pregnant Syrian hamster before and after methylation with methanolic thionyl chloride. J. roy. micro Soc. 87, 77–103 (1967 c)
Stoward, P. J.: The histochemical reactions of methanolic thoinyl chloride with the anionic groups and basic protein component of sulphated mucosubstances. J. roy. micro. Soc. 88, 119–131 (1968)
Stoward, P. J.: The mechanism of the apparent histochemical deamination of tissue proteins by sodium hypochlorite. Proc. roy. micro. Soc. 6, 16 (1970)
Stoward, P. J., Plant, M.: The histochemical deamination of proteins with sodium hypochlorite. Proc. roy. micr. Soc. 3, 21–22 (1968)
Stoward, P. J., Burns, J., Plant, M.: A tissue adhesive for paraffin sections intended for deamination with sodium hypochlorite. Histochemie 14, 212–214 (1968)
Tomasi, J. A. de: Improving the technic of the Feulgen stain. Stain Technol. 11, 137–144 (1936)
Urk, H. W. van: Het aantoonen van chlooramine en de Onderscheiding van hypochlorieten. Chem. Weekblad 26, 9–10 (1928)
Wieland, T., Vogelbach, C., Bielig, H. J.: Das Verhalten der Aminosäuren gegenüber Natriumchlorit und ihre quantitative Desaminierung mit Hypochlorit. Ann. 561, 116–123 (1949)
Wigglesworth, V. B.: Structural lipids in the insect cuticle and the function of oenocytes. Tissue Cell 2, 155–179 (1970)
Wigglesworth, V. B.: Bound lipid in the tissue of mammal and insect: a new histochemical method. J. Cell Sci. 8, 709–725 (1971)
Whistler, R. L., Schweiger, R.: Preparation of D-arabinose from D-glucose with hypochlorite. J. Amer. chem. Soc. 81, 5190–5192 (1959)
Whistler, R. L., Yagi, K.: Further application of the hypochlorite method of chain shortening in the carbohydrate series. J. Org. Chem. 26, 1050–1052 (1961)
Wright, N. C.: The action of hypochlorites on amino-acids and proteins. The effect of acidity and alkalinity. Biochem. J. 30, 1661–1667 (1936)
Author information
Authors and Affiliations
Additional information
Dedicated to Professor Dr. W. Graumann on the occasion of his 60th birthday and in honour of his contributions to histochemistry over many years both as a scientist and as an editor.
Rights and permissions
About this article
Cite this article
Stoward, P.J. A histochemical study of the apparent deamination of proteins by sodium hypochlorite. Histochemistry 45, 213–226 (1975). https://doi.org/10.1007/BF00507696
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00507696