TABLE VIII 



Pttrification of Auxin n from Urine 



(Adapted from Kogl, Haagen Smit, and Erxlohen, 1933) 



Treatment 



Acidify and extract 

 with ether 



Fractionate with 

 NaHCOs 



Extract with petro- 

 leum ether 



Extract with 

 hgroin 



hot 



Dissolve in aqueous 

 alcohol and extract 

 with benzene 



Extract with water 

 and then with 50 

 per cent methanol 



Evaporate methanol 

 and extract with 

 ether 



Dissolve in aqueous 

 alcohol and frac- 

 tionate by lead salt 

 formation 



Acidify and extract 

 with ether 



Dissolve in aqueous 

 alcohol and add 

 KOH and 

 CaCCHsCOO)., 



Heat with 1.5 per cent 

 HCl in methanol 



Distil in vacuo 



Active Fraction 



Urine Concentrate (from 



150 liters) (contains 

 about 15000 X 10^ AE) 



Recrystallize from 

 alcohol-ligroin or 

 aqueous acetone 



i 



Ether extract 

 Acid fraction 

 Residue 

 Residue 



Benzene layer 



Aqueous and meth- 

 anol extracts 



Ether extract 



Filtrate and (some- 

 times) ppt. from 

 alkaline solution 



i 

 Ether extract 



Filtrate 



Neutral fraction 



i 

 Middle fractions 

 (125°-135°) 



i 

 Crude crystallisate 

 (contains 560- 

 840 X lO" AE) 

 (i.e. yield 3-6 per 

 cent) 



Wt. 



(grams) 



87 



45 



19.7 



0.0 



3.2 



2.25 



1.2 

 0.18 



0.04 



Inactive Fraction 



Residue 



Neutral fraction 

 Petroleum extract 

 Ligroin extract 



Aqueous alcohol 

 layer 



Benzene layer 

 Aqueous layer 



Ppt. from neutral so- 

 lution and (some- 

 times) ppt. from 

 alkaline solution 



Residue 



Ppt. (colored) 



Acid fraction 



First fractions and 

 residue 



Auxin a (m. 196°) Auxin a lactone (m. 173°) 



107 



