Subject: to Dr. Peter E. Hughes, Ph. D. Biochemistry

Hi
I sent before that when I prepared plant extracts in 3% sulphosalicylic acid  
I saw the red color but what happend when I add a mixture of acid-ninhydrin 
and  acetic acid to 90 c, for 1h  and extracted the reaction mixture with 
toluene not all the color present in the toluene phase, 
I received an answer from Dr. Peter E. Hughes, Ph. D. Biochemistry that the 
color maybe the anthocyanin
Also I expect that the color maybe anthocyanin , so I prepared plant extracts 
in 3% sulphosalicylic acid (using fresh weight) and measured the anthocyanin 
using spectrophoto meter at O.D 535 and 650 , and I found that there is a 
significant differences between well water and drought treatment, so I tried 
to test the anthocyanin followed lange et al method 1971 (plant physiol,47:649-
655) using propanol-HCl-H2O with plant dry weight (because I had not fresh 
weight) and I found there was a different between the two treatments but the 
different was not significant maybe because there were  differences in the 
value of replications within the same treatment. So my questions are
1-is the heat (because I used it when I dried my samples) effect on the 
anthocyanin
2- when I extract proline form plant tissue , the ammonia produced in this way 
can react with the hydrindantin and another molecule of ninhydrin to yield a 
purple product. So why Dr. Peter E. Hughes think that the pink I noted in the 
toluene phase is due to the partial solubility of aqueous salicylate in 
toluene .
4- Do you think that anthocyanin prevent proline-ninhydrin product

Re: to Dr. Peter E. Hughes, Ph. D. Biochemistry