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Qualitative Test for Carbohydrates
Procedure: Procedure : Mix 1 ml of 5 % glucose solution with 1 ml of Nylander¶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.
Glycogen and Starch ± crystal clear but starch has a cloudy precipitate at the bottom . Lactose and Maltose ± light yellow but maltose is darker. the bismuth hydroxide formed is reduced to black metallic bismuth as shown in the following equation: equation : Bi(OH)2NO3 + KOH Bi(OH)3 + KNO3 2Bi(OH)3 (with reducing sugar and heat) 2Bi + 3O + 3H2O a test for glucose in urine using a solution containing bismuth subnitrate which forms a black precipitate in a positive reaction. darker. bottom. Galactose ± yellow orange Glucose ± dark olive green Fructose ± olive green Sucrose ± clear olive green .Nylander¶s Test In the case of alkaline solution bismuth subnitrate. reaction .
Lactose & Maltose ()(-) (L-R) Sucrose & Galactose (+)(-) (L-R) Glycogen & Starch (-)(-) (L-R) Fructose & Glucose (+)(+) (L-R) .
Add 5 drops of HCl to the 7 test tubes. fructose.Barfoed¶s Test Procedure: Procedure : Mix 1 ml of Barfoed¶s reagent with 1 ml of 5 % glucose solution . sucrose. maltose. minutes. starch and glycogen . Heat for 30 seconds and allow to stand for 10 minutes . lactose. Observe minutes. . Do the same with galactose . glycogen. solution. the changes . heat for 5 minutes . changes. galactose.
sugars . the sugars reduce less vigorously. to differentiate the two sugars. The former react. . disaccharides. Barfoed¶s test utilizes this fact for distinguishing monosaccharides from reducing disaccharides. whereas the latter do not. disaccharides . GALACTOSE In acid solution.Barfoed¶s Test Solution of cupric acetate in weak acetic acid ( Barfoed¶s ) is reduced to monosaccharides but not by disaccharides . not . POSITIVE ONLY FOR REDUCING MONOSACCHARIDES WHICH ARE: GLUCOSE. This serves therefore. vigorously. FRUCTOSE.
. but do so at a slower rate . Fructose. disaccharides undergo the same reaction. Reducing minutes.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 . anymore. 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 . Galactose. rate.
solution. Heat to boiling . with all the different types of sugar solution . carbonate .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 % sodium 10% carbonate. . Do the same boiling.
Picric Acid Test / Jaffe Reaction Some organic acids when in alkaline solutions are also reduced.Orange . Thus. 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(NO 2 ) 3 OH(NO2 C 6 H 2 OHNH 2 (NO 2 ) 2 OHNH2 (NO2 Starch and Glucose ± Colorless Glucose± Glucose. Fructose and Galactose ± Red orange Lactose and Sucrose ± Light orange Maltose . reduced . picric acid ( yellow ) is reduced to picramic acid ( mahogany red ) .
bath.Seliwanoff¶s Test Procedure: Procedure : Mix 1 ml of Seliwanoff¶s reagent in each of 6 test tubes. and lactose to the sixth . galactose to the third. Boil in water bath . maltose to the fifth. To the 1 st test tube add 1 ml of 5 % freshly prepared fructose solution . Note carefully the color produced and record the time required for the development of pink color in each of the test tubes. sucrose to the fourth. Glucose to the solution. . tubes . sixth. second. tubes .
fructose.Seliwanoff¶s Test This involves the action of resorcinol and HCl on sugar.29sec.lactose3 : 30sec. developed.maltose . sugar. a red color is developed . This is a specific test for ketose sugar.2 : 43 sec.glucose. In the presence of ketose sugar. dehydration . galactose.43sec.50sec Pink but after sucrose and fructose were very red .2 : 29 sec.maltose. compounds. 55 sec.3 : 50 sec 30 sec.46 sec.glucose . like fructose.fructose sucrose . which forms rapidly hydroxymethyl furfural as a result of dehydration. sucrose.fructose55sec.46sec. as in the case of fructose .lactose galactose . This condenses with resorcinol to form red colored compounds . Positive only for sugars having potentially free ketone group.
solution . Do the same with glycogen .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. . Add 2 drops of a very dilute aqueous solution of iodine in potassium iodide to 1 ml of starch solution. What is the color of the solution? Heat the colored solution to boiling and observe the effect. water. glycogen. temperature . Now what do you observe? Add a few drops of sodium thiosulfate solution to the cooled starch solution and note the result. effect . Allow the solution to cool back down to room temperature. result .
Prolonged heating or grinding causes the disintegration of the granules liberating the amylose which is slightly soluble in water.Iodine Test Starch raw starch is insoluble in cold water. upon heating it cleared and then upon cooling turned purple . amylopectin produces a purple color. forming an opalescent solution. upon heating it was still the same until cooling when dropped with sodium thiosulfate it also cleared. thiosulfate.black again and purpleupon dropping sodium thiosulfate . form which it can be precipitated by ethyl alcohol. and in giving with iodine a red instead of a blue color. W hen heated in water the granules swell forming a viscous hydrophillic colloidal substance. This is responsible for the deep blue color produced with iodine due to the formation of starch iodine. W ith glycogen it did not have a purple purpleblack color. it turned clear again. iodine test: starch plus lugol's iodine solution produced purple purpleblack color. Glycogen It differs from starch in being soluble in water. .
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.. it doesn't let the glucose bond together. Then sodium thiosulfate is an "inhibitor".. That is why it turned clear again . that is why it tested positive for starch. then you heat it up it is not starch anymore. called starch. 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.
seen.Mucic Acid Test Procedure: Procedure : To each test tube filled with 1 pinch of galactose for the first. crystals. glucose for the second. If no crystals appear let stand until the next lab period . then allow to cool at room temperature . Examine crystals under the microscope and draw as seen . and sucrose for the fifth add 1 ml of 5 N HNO3 . Not carefully which tubes shows water insoluble crystals . Confirm the solubility of the crystals by adding 1 ml of water to the tubes where crystals are formed . period. fructose for the fourth. formed. . temperature. HNO 3 Heat the test tubes in boiling water for 1 hour. lactose for the third.
Mucic Acid Test Positive only for galactose due to the formation of an insoluble mucic acid crystals when heated with concentrated nitric acid. Galactose. forms mucic acid. . an isomer of saccharic acid. Mucic is insoluble and forms characteristic sandy crystals which serve to identify galactose . on being oxidized with HNO3. Galactose . galactose.
.White precipitate settled .Water insoluble crystals Galactose ± white precipitate is present and the oil is floating Water soluble crystals Lactose . settled.
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