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Chlorocruoroporphyrin: a Simple Method of Preparation

Published online by Cambridge University Press:  11 May 2009

Gilbert Y. Kennedy
Affiliation:
Cancer Research Laboratory, University Field Laboratories, Lodge Moor, Sheffield 10

Abstract

Munro Fox (1926, Addendum, p. 219) described a method of preparation of chlorocruoroporphyrin from the blood of the marine polychaete worm Spirographis. This was based upon the very troublesome method of Nencki & Zaleski (1900), which is time-consuming and wasteful. Furthermore, Spirographis is not easily obtainable in this country. Lemberg & Parker (1952) started with protoporphyrin, and by careful oxidation with potassium permanganate in acetone obtained a solution which gaVe mixed crystals of chlorocruoroporphyrin and diformyldeuteroporphyrin esters. Purification was by fractional chromatography. The procedure to be described here is simple and rapid, and could easily be carried out in one day by a biologist requiring a small sample of the porphyrin. The two methods mentioned above would not come into this category.

This work was done at the Plymouth Laboratory, and I am very grateful to Mr T. R. Tozer of that laboratory for so kindly obtaining the animals for me and extracting the blood.

The polychaete worms Myxicola infundibulum, Sabella pavonina and Branchiomma vesiculosum are all readily obtained from the muddy shores of the Salcombe estuary, especially on the Salstone. Blood was taken directly from the vessels of about 100 assorted worms by syringe, and squirted as it was removed from the worms into a mixture of peroxide-free ether (3 parts) and ‘Analar’ glacial acetic acid (1 part). After all the blood had been added, the mixture was well shaken and allowed to stand in the ice-chest for 30 min. The acetic acid was then washed out with distilled water, the first few washings containing a little sodium acetate to avoid washing out of the pigment, and the remaining solution of chlorocruorohaematin dried roughly by filtering through ether-soaked paper into a distilling flask and evaporated to dryness. 100 ml. of a 50% solution of hydrazine hydrate in glacial acetic acid were then added, and the mixture heated on a water-bath at 90° C. for 10 min.

Type
Research Article
Copyright
Copyright © Marine Biological Association of the United Kingdom 1953

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References

REFERENCES

Fox, H. Munro 1926. Chlorocruorin: a pigment allied to haemoglobin. Proc. Roy. Soc., B, Vol. 99, pp. 199220.Google Scholar
Ľemberg, R. & Parker, J. 1952. Porphyrins with formyl groups. II. Preparation of chlorocruoroporphyrin and diformyl deuteroporphyrin. Austral. J. Exp. Biol. Med. Sci., Vol. 30, pp. 163170.CrossRefGoogle Scholar
Nencki, M. & Zaleski, J. 1900. Untersuchungen über den Blutfarbstoff. Z. physiol. Chem., Bd. 30, 384pp.CrossRefGoogle Scholar