m

    



 

a
a 
a


 


à


` Test for the presence of guanidine
à


` Test for the presence of guanidine

V

à


` eagents: ` @echanism:
` Ʉ-naphthol
` NaOH
` *NaOCl
+ NaOH, pH

Ô


 

à



 
 



|
  |


 
@ 

  
` Àor compounds containing a phenolic hydroxy group
` Amino acid: Tyrosine

` Compound must be
validated as protein/
amino acid to confirm
presence of tyrosine
@ 

  
` @illonǯs eagent: @ercuric ion in acid
` @echanism:
` Tyrosine + @illonǯs eagent = complex
@ 

  
` Complex treated with Nitrous Acid (NaNO2) yields a
pink-red solution
ÔV  V
ÔV  V
` uoiling concentrated nitric acid reacts with tyr, trp
and phe to yield yellow products.
` This reaction involves the nitration of benzene nucleus
in alkaline medium. As a result, amino acids that
contain aromatic nucleus undergo this reaction.
` Aromatic AAs form yellow nitro derivative on heating
with concentrated nitric acid, the salts of this
derivative are orange.
ÔV  V
V V àV
 V 
ÔV  V
` £sing 65% nitric acid the aromatic rings of amino
acids like tyrosine and tryptophan are nitrated. The
nitro derivate show an intensely yellow color. uecause
nearly all proteins contain aromatics it is taken as a
protein-test either.

` The yellow stains on the skin caused by nitric acid are

the result of the xanthoproteic reaction. The epidermis
cells of the skin contain aromatic proteins.
 
V
  V
` Triketohydrindene hydrate, commonly
known as ninhydrin reacts with amino
acids to form a purple colored imino
derivative. This derivative forms a
useful test for amino acids, most of
which are colorless.
` Ninhydrin is a powerful oxidizing
agent which reacts with all amino acids
between pH 4-8 to produce a purple-
colored compund
  V
` V color reaction given by amino acids
and peptides on heating with the
chemical ninhydrin
` The amino acids proline and
hydroxyroline also reacts but produces
a yellow color.
` Ninhydrin (triketohydrindene
hydrate) is an oxidating agent which
leads to the oxidative deamination of
alpha-amino groups. It is very
important for the detection and the
quantitative analysis of amino acids.
  V
` Ninhydrin also reacts with primary
amines however the formation of
carbon dioxide is quite diagnostic for
amino acids.
` Vlpha amino acids yield a purple
substance that absorbs maximally at
570 nm. Imino acids (proline) yield a
yellow product (absorption maximum
440 nm).
  V
Ʉ-amino acid + 2ninhydrin ‰
CO2 + aldehyde + final complex(purple) +
3H2O
Ninhydrin, which is originally yellow,
reacts with amino acid and turns deep
purple color that is detected in this
method.
  

` resence of peptide bonds is detected by performing a
chemical test named biuret test.

` The uiuret eagent is made of sodium hydroxide and

copper sulfate. The blue reagent turns violet in the
presence of proteins, and changes to pink when
combined with short-chain polypeptides.
` Cupric ion in an alkaline medium forms a violet
coloured complex with peptide bond nitrogens of
peptides and proteins.

` The reaction is so named biuret(NH2CONHCONH2)

formed by condensation of two molecules of urea,
when heated at 180C, also answers this test. The
minimum requirement for a positive test is the
presence of 2 peptide bonds in the molecule.
`dt yielded a positive result for
saliva thus, saliva contains
proteins.
`dt yielded a negative result for
glycine.
à 

     

` The bromine water test is an example of an addition
reaction.(A reaction in which a small molecule adds on
across a double bond).

` The decoloration of a solution of bromine in water is

an analytical test for the presence of alkenes:

CH2=CH2 + ur2 ü urCH2-CH2ur

`Alkenes are able to undergo
addition reactions because
they contain a double bond.
`They decolourise because
they are unsaturated and
have a carbon=carbon double
bond
  
` Àormation of pinkish
layer is the positive
result.

` Tryptophan is positive
under the bromine water
test
 Pauly Reaction
  

 |   

‰à    
 

 
 |
 


|  
 

 |  
 

àà 
|! || 



  
|
 |



àamples

à  
 


  
|
 

"

 
|
 

# |  |
 
à 
|

"


 Îead Acetate Reaction
Î | 

 | 
 


à ||
  
  | 
| 

| ! 

|   ! 

  |  
 ||
 
àà| 

  

  |



àamples

à  
" |
 
 |
 
 

  

 |
  | 

# |  |
 
References
$ $$
$$V

V
$  % |
&
'()('|
 $ | !$ *
 |"Î (Î 
| 
( 

(  ( 

$
$ $$
$$
Ô
Ô 
   % |
&
'()('|
$ 
$%  (|  |  | 

   (| 
(| 
$
$ $$
$$

$  % |
&
'()('|
$! | $  ()$