General scheme

1- biuret test
Test Observation Result
biuret test Violet color Solution is Protein
(3ml sample +1.5 ml biuret (albumin, casein, gelatin)
reagent )
Blue color Solution is (Carbohydrate)

Rosy pink Solution is peptone

If biuret test give no violet color follow carb scheme

and if it give violet color follow protein scheme
If biuret test give no violet color follow carbohydrate scheme
Test Observation Result
1-Iodine test Dark blue color The sample is starch (polysaccharide)
(2 ml of sample + 2 drops
of iodine) No blue color The sample is not starch

2-Fehling’s test Red ppt. The sample is reducing sugar (glucose,

(2 ml of sample + 1ml fructose ,lactose)
fehling’s A + 1ml fehling’s No ppt. The sample is sucrose (non-reducing sugar)
B) then heat on water
bath

3-Barfoed’s Red ppt. The sample may be glucose or fructose

To 1 mL of carbohydrate (monosaccharide)
+ 5 mL of Barfoed reagent
No red ppt. The sample is reducing disaccharide (lactose)
heat on water bath

4-Seliwanoff’s test Cherry red color The sample is fructose

(1 ml of sample+ 3_5 ml
seliwanoff’s reagent) Then Negative result (no change The sample is glucose
heat. in color)
Pale red color The sample is sucrose
(Confirmatory for sucrose)
If biuret test give rosy to violet color follow protein sceme :
Test Observation Result
1-Heat coaulation test white Coagulation The sample is albumin
7ml sample then heat
No white coagulation The sample is not albumin (may
be casein or gelatin)
2-xanthoprotei test yellow or orange ppt The sample is albumin or casein
2 mL of protein solution+
1 mL conc.HNO3
(SLOWLY ON THE SIDE WALL) ,heat then
No Yellow color The sample is gelatin
cool then add 5 mL of 40% NaOH
(No color change)
(this test is used to differentiate between
casein and gelatin and confirmatory for (yellow or orange color) The sample is peptone
albumin and peptone)

4-Rosenheim’s Test Violet Ring The sample is Albumin, Casein

3 mL of the protein solution, 2 drops of or Peptone
Rosenheim reagent, mix well then add 3
mL of conc.H2SO4 (run slowly on the wall)
No Violet Ring The sample is Gelatin
(this test is confirmatory for albumin)

Note :
Casein is the only sample that give effervescence by addition of nitric acid.
 Starch
Test Observation Result
1-biuret test Blue color The sample is carbohydrate
(3ml sample +1 ml biuret
reagent )
2-Iodine test Dark blue color The sample is starch (polysaccharide)
(2 ml of sample + 2 drops of
iodine)
 Sucrose
Test Observation Result
1-biuret test Blue color The sample is carbohydrate
(3ml sample +1 ml
biuret reagent )
2-Iodine test No blue color The sample is not starch
(2 ml of sample + 2
drops of iodine)
3-Fehling’s test No ppt. The sample is sucrose (non-reducing sugar)
(2 ml of sample + 1ml
fehling’s A + 1ml
fehling’s B) then heat
on water bath

4-Seliwanoff’s test Pale red color The sample is sucrose

(1 ml of sample+ 3_5 (Confirmatory for sucrose)
ml seliwanoff’s
reagent) Then heat.
 Lactose
Test Observation Result
1-biuret test Blue color The sample is carbohydrate
(3ml sample +1 ml
biuret reagent )
2-Iodine test No blue color The sample is not starch
(2 ml of sample + 2
drops of iodine)
3-Fehling’s test Red ppt. The sample is reducing sugar
(2 ml of sample + 1ml (glucose, fructose ,lactose)
fehling’s A + 1ml
fehling’s B) then heat
on water bath

4-Barfoed’s No ppt The sample is reducing

To 1 mL of disaccharide (lactose)
carbohydrate + 3 mL of
Barfoed reagent heat
on water bath
 Fructose
Test Observation Result
1-biuret test Blue color The sample is carbohydrate
(3ml sample +1 ml
biuret reagent )
2-Iodine test No blue color The sample is not starch
(2 ml of sample + 2
drops of iodine)
3-Fehling’s test Red ppt. The sample is reducing sugar (glucose, fructose
(2 ml of sample + 1ml ,lactose)
fehling’s A + 1ml
fehling’s B) then heat.

4-Barfoed’s Red ppt. The sample may be glucose or fructose

To 1 mL of (monosaccharide)
carbohydrate + 3 mL of
Barfoed reagent then
heat .

5-Seliwanoff’s test Cherry red color The sample is fructose

(1 ml of sample+ 3_5
ml seliwanoff’s
reagent) Then heat.
 Glucose
Test Observation Result
1-biuret test Blue color The sample is carbohydrate
(3ml sample +1 ml
biuret reagent )
2-Iodine test No blue color The sample is not starch
(2 ml of sample + 2 drops
of iodine)
3-Fehling’s test Red ppt. The sample is reducing sugar
(2 ml of sample + 1ml (glucose, fructose ,lactose)
fehling’s A + 1ml fehling’s
B) then heat on water
bath.
4-Barfoed’s Red ppt. The sample may be glucose or fructose
To 1 mL of carbohydrate (monosaccharide)
+ 3 mL of Barfoed reagent
heat on water bath.
5-Seliwanoff’s test Negative result The sample is glucose
(1 ml of sample+ 3_5 ml (no change in color)
seliwanoff’s reagent)
Then heat.
 Albumin
Test Observation Result
1-biuret test Violet color Solution is Protein
(3ml sample +1 ml biuret reagent ). (albumin, casein,
gelatin)

2-Heat coaulation test white Coagulation The sample is albumin

7ml sample then heat
3-xanthoprotei test yellow or orange ppt The sample is albumin
( 2 mL of protein solution+
1 mL conc. HNO3
(SLOWLY ON THE SIDE WALL) ,heat A
Yellow color or precipitate is formed ,
cool then add 5 mL of 40% NaOH .
(this test is confirmatory for albumin)

4-Rosenheim’s Test Violet Ring The sample is albumin

3 mL of the protein solution, 2 drops of
Rosenheim reagent, mix well then add 3
mL of conc. H2SO4 (run slowly on the wall)
(this test is confirmatory for albumin)
 Casein
Test Observation Result
1-biuret test Violet color Solution is Protein
(3ml sample +1 ml biuret reagent ) (albumin, casein,
gelatin)

2-Heat coaulation test No white coagulation The sample is not

7ml sample then heat albumin (may be casein
or gelatin)
3-xanthoprotei test yellow or orange ppt The sample is Casein
( 2 mL of protein solution+
1 mL conc. HNO3
(SLOWLY ON THE SIDE WALL) ,heat A Yellow
color or precipitate is formed , cool then add 5
mL of 40% NaOH .

4-Rosenheim’s Test Violet Ring The sample is Casein

3 mL of the protein solution, 2 drops of
Rosenheim reagent, mix well then add 3 mL of
conc. H2SO4 (run slowly on the wall)
(this is confirmatory test(
 Gelatin
Test Observation Result
1-biuret test Violet color Solution is Protein
(3ml sample +1 ml biuret reagent ) (albumin, casein,
gelatin)

2-Heat coaulation test No white coagulation The sample is not

7ml sample then heat albumin (may be casein
or gelatin)
3-xanthoprotei test No Yellow color The sample is Gelatin
( 2 mL of protein solution+ (No color change)
1 mL conc. HNO3
(SLOWLY ON THE SIDE WALL) ,heat A
Yellow color or precipitate is formed ,
cool then add 5 mL of 40% NaOH .

4-Rosenheim’s Test No Violet Ring The sample is Gelatin

3 mL of the protein solution, 2 drops of
Rosenheim reagent, mix well then add 3
mL of conc. H2SO4 (run slowly on the wall)
(this is confirmatory test(