130 



PREPARATION AND CRYSTALLOGRAPHY OF HEMOGLOBINS. 



TABLE 34. Effects of the condition of the blood and temperature upon 

 crystallization according to Uhlik. 



Uhlik also notes that Pregl found that a thrice-crystallized hemoglobin 

 appeared as hexagonal crystals. Crystals of reduced hemoglobin have 

 been prepared and described by a number of investigators, as stated in 

 previous pages. 



The last hemoglobin to be obtained in crystalline form, excluding our 

 own preparations, was prepared by Bardachzi (Zeit. f. physiolog. Chemie, 

 1906, XLIX, 465) from the blood of the sea-tortoise (Thalassochclys corti- 

 cata). The blood was centrifugalized, the corpuscles mixed with water, 

 and then set aside for several hours at 50. The solution was then filtered, 

 one-fifth volume of alcohol added to the filtrate, and the mixture placed in 

 an ice-chest. Crystallization occurred quickly and abundantly in the form 

 of plates. The crystals were soluble with difficulty in cold water. 



For the purpose of analysis the crystals were dissolved in water at 40, 

 and after cooling one-seventh volume of alcohol was added, and crystalliza- 

 tion obtained as before. The crystals were then centrifugalized off and 

 dried in vacuum. The mean values of the elementary analyses were 



C54-77He-99Ni7. 0780-38^60-41 



The absence of phosphorus is striking, since previous observers failed to 

 obtain hemoglobin free from phosphorus from bloods that contain nucleated 

 erythrocytes. The optical investigation by means of the Hiifner spectro- 

 photometer showed decided agreement with the blood-coloring matter of 

 such other animals as have been closely investigated up to this time. The 

 average quotient was e' : e = 1.561, while Hiifner found the quotient to be 

 1.578. The calculation of the extinction coefficients and quotients of hemo- 

 globin and methemoglobin agreed, he states, with those of other oxyhemo- 

 globins and methemoglobins, so (hat the coloring matter of the blood of the 

 tortoise, Bardachzi holds, is identical with that of mammals. 



Abderhalden and Medigreceanu (Zeit. f. physiol. Chemie, 1909, LXIX, 

 165) report their preparation of crystals of goose hemoglobin free from 

 phosphorus. 



