Ajara Mahavidyalaya,ajara
Department of Zoology

B.Sc.II (Biochemistry)

Detection of Unknown solution:-

Sr.No
Test
Observation
Inference
1
2 ml O.S. + 1ml NaOH + 2drops of CuS04
Violet colour

Protein Present.

No violet colour
Carbohydrate Present.
2
2 ml O.S. + Iodine solution.
Blue colour

Starch present.

No Blue colour
Saccharides Present.

C.T. For Saccharides :-

Sr.No
Test
Observation
Inference
1
2 ml O.S. + 2 drops of Molash Reagent,mix well.Add this to inclined test tube 2ml conc.H2So4 drop by drop.
Brown ring at the junction of two solutions
Saccharides Present.
2
Tomers Test :-
2 ml CuS04 + 2ml O.S.Mix well+ 2ml NaOH-Blue coloured ppt then boil.
Yellow ppt.
Fructose or lactose present.


3
Fehlings Test:-
1ml fehling solution boil – no change in color + O.S. boil

Breek Red ppt.
Fructose or Lactose Present.
4
Bendedict's Test:-
1 ml. Bendedict's Reagent +0.5ml. O.S. -Boil and allow the tube stand for 10 minutes.
Yellow colour chainges to orange.
Fructose or Lactose Present.

If all the above test are nigative – Then Sucrose Present.

Sr.No
Test
Observation
Inference
5
C.T. For Sucrose:-
2ml. O.S. + 1 ml. Dil.HCL, Boil And Cool. Nautralize With NaOH +2 ml CuS04 Boil.
Yellow or Red ppt.
Sucrose Confermed.
6
C.T. For Fructose and Lactose:-
Seliwanoff Test :-
2 ml O.S.+Seliwanoff reagent Boil for two minutes.
Red Colour
Fructose Confermed.
No Red colour
Lactose confermed.
7
Osazone Test :-
5ml O.S.+3 drops of acetic acid+pinch of Phenyl Hydrozine Hydrochloride + Sodium acetate mix well.Keep in waterbathfor 20 to 30 min,cool and observe under microscope.
Observe Osazone crystals.

Fructose:-Needle Shaped.
Fructose Confermed.
Lactose :-Cotton ball or powder puff shaped.
Lactose confermed.
Sucrose :- No Crystals
Sucrose Confermed.


C.T.For Starch:-
Sr.No.
Test
Observation
Inference
1
3 ml NaOH + O.S. Boil (Jelly is formed) + 1 drop of Iodine solution.
No Blue colour.
Starch Present.
2
Add Conc. HCL to above solution to nutralize NaOH.
Blue colour appears
Starch confermed.
3
1 ml O.S.+2 drops of Conc.HCL boil for 3 minutes and add to it 1 ml 10% NaOH + 2 drops of CuS04 ----Boil
Blue coloured ppt. Turns Yellow to break red.
Starch Present and confermed.

C.T. For Proteins :-

Sr.No.
Test
Observation
Inference
1
Xanthoprotein Test:-



a) 2 ml O.S. + 1ml conc.HNO3 Boil and cool
White ppt. Turns Yellow on heating
Protein Present.
b) Add NaOH in above test tube excess
Yellow colour turns orange
Protein Present.
2
Biuret Test :-
3 ml O.S.+ 3 ml NaOH mix well + 3 drops of CuS04
Violet or Pink Colour
Protein Present.
3
Cystine Test :-(Sulpher reaction test)
3 ml O.S. + 5 drops Lead accetate ppt.+ NaOH drop by drop until ppt. Dissolves,then boil.

Darkning of the solution due to formation of the Lead sulphide.
Protein Present.
4
Ninhydrine test:-
1 ml O.S.+ 10 drops of Ninhydrine solution boil and cool.
Faint blue colour develops
Protein Confermed.
5
Alcohol Test:-
1 ml O.S.+ few drops of absulate alcohole.
White ppt.Forms
Protein present and Confermed.


Result :- Given sample of the solution contains____________.

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