276 



ADVENTURES IN RADIOISOTOPE RESEARCH 



We determined the blood phosphorus of the laying hen denoted as I , 

 the result being seen from Table 3. The blood phosphorus of another 

 lien is discussed on p. 274. 



Table 3. 



P-CONTENT OF HeNS BlOOD 



iu plasma 



mgm "/(, 



mgin % 



ill oorpuscles iu blood 



Phospliatide P 



Inorganic P 



Total acid soluble P 

 Rest (Protein) P . . . 



20.7 



21.3 



16.8 



In the blood of non-laying hens(J> after 24 hours fasting an average 

 phosphatide P content of 16.8 mgm% was found the total plasma P 

 amounting to 13 mgm%, the plasma inorganic P to 4.6 mgm%. 



EXPERIMENTAL METHODS 



The yolk wus dried by adding ice cold acetoii, the dry yolk was carefully 

 pulverised and the powder obtained shaken for 15 min with 150 ce. ether, the 

 last mentioned procedure being repeated four times using fresh ether. The ether 

 was than carefully evaporated, the residue taken up with dry ether, the latter 

 removed by evaporation, this time in a Kjeldahl flask, and the residue ashed. 



The phosphatides of plasma, corpuscles and total blood were extracted b_\- an 

 ether-alcohol mixture after Bloor. The extract was several times carefully evapo- 

 lated to dryness and taken up with ether or petrol ether. The residue of the first 

 extraction was treated with trichloracetic acid (10 cc. of 10% solution for each 

 cc. of blood) and from the filtrate obtained the inorganic P precipitated as ammo- 

 nium magnesium phosphate; the esters present in the filtrate were hydrolised 

 and the phosphate produced by the hydrolysis of the esters precipitated as ammo- 

 nium magnesium phosphate. Though the extraction and the neutrahsation of the 

 acid solution were both carried out at — 9°, some of the inorganic phosphorus 

 present may be due to decomposition of the esters and we therefore gave in the 

 table only the total acid soluble phosphorus present in the corpuscles which 

 includes both the inorganic and the ester phosphorus. 



The liver was minced, dried in vacuo at room temperature, pulverised, 

 dried again in vacuo and extracted with ether-alcohol (1:3), the latter being 

 left to boil for 15 sec. In one case we extracted with ether alone to com- 

 pare the active P content of the ether soluble phosphatides such as lecithin 

 with that of the total phosphatides. The atad soluble P was extracted from 

 the dried liver powder by treatment with cold ( — 10° to — 15°) solution of 

 trichloracetic acid, first with a IO'^/q solution for 10 min and than twice 

 w ith a 5 ^/q solution each for 5 min. The inorganic and organic constituents 

 of the acid soluble phosphorus were separated as stated above. The P con- 

 tent and activity of the residue obtained after extraction of the phosphatides 

 and the acid soluble P was also investigated. 



1 H. M. DvKK and I. H. Hoe, J. Niitrit. 7, (523 (1<)34). 



