Studies With C^^-Iabeled Ethylene 



759 



Aqueous extract 



Effluent -<- 

 (discard) 



Adsorb on Dowex-2 



(CO3-) 



Elute with O.IAT 

 formic acid + O.OSiV 

 NH4CHO0 



-^ Eluate I 



(chromatograph) 



Elute with 0.15iV 

 NaCl + 0.05N HCl 



> Eluate II 



(chromatograph) 



Fig. 3. Fractional elution scheme of water-soluble cotton metabolites. 



tion from an anion exchange column as shown in Figure 3. The radio- 

 active fractions were neutralized, reduced in volume, and chromato- 

 graphed in two solvent systems. 



Inspection of the histograms presented in Figure 4 shows at least 

 two radioactive fractions were obtained by elution from the anion 

 exchanger. The activity in the area of the second peak is almost 14 

 times that of the first peak. Strong adsorption on the column denoted 

 a fairly acidic compound. 



Chromatograms of the fractions, sprayed with Hanes and Isher- 

 wood's reagent (6), gave a positive test for phosphates and the dupli- 

 cate radioautograms (Figure 5A, Sh 39 and 40) showed that the main 

 radioactive spots corresponded with the molybdenum blue areas, but 

 with two additional spots above them. The main metabolite from the 

 anion exchanger was chromatographed in two solvent systems and 

 the respective radioautograms are shown in Figure 5B, Sh 41 and 42. 

 The same fraction after purification was chromatographed in Ban- 

 durski and Axelrod solvents (3) and sprayed for phosphate esters 

 (Figure 5B, Sh 43 and 44). The presence of phosphate-containing 

 compounds was demonstrated in all cases and they corresponded with 



