Activity 6-B

|

 




ylander¶s Test

` Procedure::
Procedure
` Mix 1 ml of 5 % glucose solution
with
1 ml of ylander¶s reagent
reagent..
` Heat for 5 minutes in a boiling
water
bath and note the result
result..
` Do the same with galactose
galactose,,
fructose, sucrose, lactose,
maltose, starch and glycogen
glycogen..
ylander¶s Test
` a test for glucose in urine using
` In the case of alkaline a solution containing bismuth
solution bismuth subnitrate which forms a black
precipitate in a positive
subnitrate, the bismuth reaction..
reaction
hydroxide formed is ` Lactose and Maltose ± light
reduced to black yellow but maltose is darker
darker..
metallic bismuth as ` Glycogen and Starch ± crystal
shown in the following clear but starch has a cloudy
precipitate at the bottom
bottom..
equation::
equation
` Galactose ± yellow orange
` Bi(OH)2O3 + KOH ` Glucose ± dark olive green
ĺBi(OH)3 + KO3
` Fructose ± olive green
` 2Bi(OH)3 (with ` Sucrose ± clear olive green
reducing sugar and
heat) ĺ 2Bi + 3O +
3H2O
Lactose & Maltose (-
)(-)
(L-R)
Sucrose & Galactose (+)(-)
(L-R)

Glycogen & Starch (-)(-) Fructose & Glucose (+)(+)

(L-R) (L-R)
Barfoed¶s Test
` Procedure::
Procedure
` Mix 1 ml of Barfoed¶s reagent with 1
ml of 5 % glucose solution
solution..
` Heat for 30 seconds and allow to
stand for 10 minutes
minutes..
` Do the same with galactosegalactose,,
fructose, maltose, lactose, sucrose,
starch and glycogen
glycogen..
` Add 5 drops of HCl to the 7 test
tubes, heat for 5 minutes
minutes.. Observe
the changes
changes..
Barfoed¶s Test
` Solution of cupric    


acetate in weak acetic 

 

acid ( Barfoed¶s ) is
reduced to ` In acid solution, the
sugars reduce less
monosaccharides but
vigorously.. Barfoed¶s
vigorously
not by disaccharides
disaccharides.. test utilizes this fact
This serves therefore, for distinguishing
to differentiate the two monosaccharides
sugars.
sugars . from reducing
@
   disaccharides..
disaccharides The

 former react,
 
  
whereas the latter do
not..
not
` Reducing
monosaccharides are
oxidized by the copper
ion in solution to form a
carboxylic acid and a
reddish precipitate of
copper (I) oxide within
three minutes
minutes.. Reducing
disaccharides undergo
the same reaction, but do
so at a slower rate
rate..
` Galactose, Fructose,
Glucose and Sucrose had
red precipitate but after
adding HCl to all of the
samples except the
glucose which wasn¶t
added HCl had faded
back to its skyblue color
without the red
precipitate anymore
anymore..
Glucose
Picric Acid Test
` Procedure::
Procedure
` To 1 ml of 5 % glucose
solution add 1 ml of saturated
picric acid solution and about
5 drops of 10 10%% sodium
carbonate..
carbonate
` Heat to boiling
boiling.. Do the same
with all the different types of
sugar solution
solution..
 Picric Acid Test / Jaffe
Reaction
` Some organic acids when in alkaline solutions are also
reduced.. Thus, picric acid ( yellow ) is reduced to picramic
reduced
acid ( mahogany red ) .
` It is the basis for one of the quantitative blood sugar
methods widely used for a time and still used to some
extent..
extent
` C 6 H 2 OH(O
OH(O22 ) 3 ĺ C 6 H 2 OHH
OHH22 (O
(O22)2
` Starch and Glucose
Glucose±± Colorless
` Glucose, Fructose and Galactose ± Red orange
` Lactose and Sucrose ± Light orange
` Maltose - Orange
Seliwanoff¶s Test
` Procedure::
Procedure
` Mix 1 ml of Seliwanoff¶s reagent in each of
6 test tubes.
tubes .
` To the 1 st test tube add 1 ml of 5 % freshly
prepared fructose solution
solution.. Glucose to the
second, galactose to the third, sucrose to
the fourth, maltose to the fifth, and lactose
to the sixth
sixth..
` Boil in water bath
bath..
` ote carefully the color produced and
record the time required for the
development of pink color in each of the
test tubes.
tubes .
Seliwanoff¶s Test
` This involves the action ` Positive only for sugars
of resorcinol and HCl having potentially free
ketone group, as in the
on sugar
sugar.. In the case of fructose
fructose..
presence of ketose
sugar, like fructose, a ` sucrose - 46
sucrose- 46sec,fructose
sec,fructose--
55sec,glucose
55 sec,glucose-- 2 : 29
29sec,
sec,
red color is developed
developed.. galactose-- 2 : 43
galactose 43sec,lactose
sec,lactose--
This is a specific test 3 : 30sec,maltose
30 sec,maltose-- 3 : 50
50sec
sec
for ketose sugar, which
` Pink but after sucrose and
forms rapidly fructose were very red
hydroxymethyl furfural
as a result of
dehydration..
dehydration This
condenses with
resorcinol to form red
colored compounds
compounds..
Iodine Test
` Procedure::
Procedure

` Prepare starch solution by mixing thoroughly 2

grams of starch in 10 ml of water and then
pouring this mixture in 100 ml of boiling water
water..

` Add 2 drops of a very dilute aqueous solution

of iodine in potassium iodide to 1 ml of starch
solution..
solution

` What is the color of the solution? Heat the

colored solution to boiling and observe the
effect..
effect

` Allow the solution to cool back down to room

temperature.. ow what do you observe?
temperature

` Add a few drops of sodium thiosulfate solution

to the cooled starch solution and note the
result.. Do the same with glycogen
result glycogen..
Iodine Test
` Starch

raw starch is insoluble in

cold water. W hen heated in water
the granules swell forming a
viscous hydrophillic colloidal
substance. Prolonged heating or
grinding causes the disintegration
of the granules liberating the iodine test: starch plus lugol's
amylose which is slightly soluble iodine solution produced purple-
purple -
in water. This is responsible for black color, upon heating it
the deep blue color produced with cleared and then upon cooling
iodine due to the formation of
turned purple-
purple - black again and
starch iodine; amylopectin
produces a purple color. upon dropping sodium thiosulfate
thiosulfate,,
it turned clear again. W ith
` Glycogen glycogen it did not have a purple-
purple -
black color, upon heating it was
It differs from starch in
being soluble in water, forming an
still the same until cooling when
opalescent solution, form which it dropped with sodium thiosulfate it
can be precipitated by ethyl also cleared.
alcohol; and in giving with iodine
a red instead of a blue color.
` well the iodine is a STARCH test

so you put it in at first and there is starch right? so the

positive test is blue/black..

then you heat it up it is not starch anymore, it is glucose

starch is a long chain of glucose macromolecules. so
that is why it tested negative (it was clear)

then it cooled so the glucose macromolecules bonded

up together again in a long chain, called starch. that is
why it tested positive for starch..

Then sodium thiosulfate is an "inhibitor", it doesn't let

the glucose bond together. That is why it turned clear
again
Mucic Acid Test
` Procedure::
Procedure
` To each test tube filled with 1 pinch of galactose for the first,
glucose for the second, lactose for the third, fructose for the
fourth, and sucrose for the fifth add 1 ml of 5  HO3
HO 3 .
` Heat the test tubes in boiling water for 1 hour, then allow to
cool at room temperature
temperature..
` If no crystals appear let stand until the next lab period
period..
` Examine crystals under the microscope and draw as seen
seen..
` Confirm the solubility of the crystals by adding 1 ml of water
to the tubes where crystals are formed
formed..
` ot carefully which tubes shows water insoluble crystals
crystals..
Mucic Acid Test
` Positive only for galactose
due to the formation of an
insoluble mucic acid
crystals when heated with
concentrated nitric acid.
` Galactose , on being
Galactose,
oxidized with HO3, forms
mucic acid, an isomer of
saccharic acid. Mucic is
insoluble and forms
characteristic sandy
crystals which serve to
identify galactose
galactose..
` Water insoluble crystals

Galactose ± white precipitate is present and the oil

is floating

` Water soluble crystals

Lactose - White precipitate settled

settled..
ED `