Sp 2) sr. ALEXIUS COL & Serene rere errr] Nam Score: Course, Year & Section:_Date performed: _ tea ee F CoO BAG Activity 9: Test for Carbohydrates Objectives tthe end of the experiment, student should be able to: : ‘* Idontify diferent the test for carbohydrates ‘Differentiate the diferent classes of carbohydrates Procedure ‘A. lodine Test for Polysaccharides 1. Perform this procedure using starch, glucose, sucrose, galactose, fructose and lactose . Run a blank test using distilled water. 2. Place 3ml of starch solution in each of two test tubes. To one, add several drops of 40% NaOH, ‘Add a drop of very dilute iodine solution to each test tube, Note the color. Heat the tube in which the color developed. Coo! and note any change. Compare withthe blank tube. ‘3. Perform the iodine test on starch powder. Note the result. Compare the result with that ofthe starch solution. 4. Using other polysaccharides, perform the test by adding oné drop of iodine solution to SmL of 1% solution of carbohydrate. Observe and record your results. B. Moliseh Test 1. To three separate test tubes, place 0.SmL of 1.0% solutions of glucose, sucrose, and starch respectively 2. Add 2mL water and 2 drops of Molsh Reagent. ‘3. Mix each well. Inctine each tube and slowly add 3mL. concentrated sulfuric acid on the side of the. tube to form a layer below the sugar solution. 4. A purple ring at the junction is indicative of a carbohydrate, 4 ©. Osazone! Phenylhydrazine Test 1. Inseven separate test tubes, add 1 ml of 1.0% solutions of glucose, fructose, galactose, lactose, and sucrose and label propery 2. Ad tmL of phenythydrazine reagent. Mix the contents well and stopper with cotton loosely. 3. Putall the tubes at the same time in a large beaker of boiling water. Note the time of immersion ‘and the time of formation of each osazone, 4. Shake the tube occasionally to avoid supersaturation. 5. Remove the tubes after 30 minutes and cool. Note the sugars that form osazones only after cooling. 6. Examine the crystals under low power microscope. The crystals may be washed several times With distilled water before examination. Better crystals are obtained when allowed to cool in a water bath, 7. Draw the crystals you obtained, D. Benedict's Test 1. Prepare 0.5% and 2.0% glucose, fructose, sucrose and starch Biochemistry Manual & ges® en pppep a 1 2 is ‘To ImL reagent, add exactly 2 drops ofthe sugar solution. Mix well ‘Bot vigorously fortwo minutes (oF place in boing water bath for 3 minutes). ‘Alow the folution to cool spontaneously. Do not hasten cooling by immorsion in cold water. Barfood's Test 7 1. Use 1.0% solution ofthe following sugars for this test: glucose, sucrose, and fructose. To ‘mL of the reagent, add two drops of the sugar solution, 2. Heat to boiling for 30 seconds. I no precipitate form, allow the tubes to stand for about 15 ‘minutes and examine 3. _To.compare the reactions of monosaccharides and disaccharides, plece the tubes at the ‘Same time in boiling water bath. Note the time when signs of reduction appear in each tube, 4. Continue boiling for 15 minutes and remove the tubes fram the bath, Note the amounts of Brecipitate in each tube after standing for 18 minutes. 8. Record all observations, Soliwanott's Test 1 Use 0.1% and 1.0% solutions of glucose, fructose and sucrose for this 2. ToSmL of the reagent, place tml of the sugar solution. 3. Place them in a boling water bath for exactly 1 minute. Note the results. 4. Continue heating and observe the color change at one minute intervals for 4 minutes. Run a blank test, Bial's Test Use 1.0% solutions of glucose, fructose and galactose for thi tet ‘To Smt of Bias reagent, add 2mL of the sugar solution. . Heat until i just begins to bol. Abluegreen color forming at once or on cooling indicates a pentose. Kelonexoses and methyl pentoses produce orange or olive-green colored solutions. Tollen’s Pholoroglucinol Test Use 1.0% solutions of glucose, fructose and galactose for this test To 1 ml of each of the sugar solution, add 1 mL of concentrated hydrochleic acid and 2-9 drops of Tollen’s Phloroglucinol reagent. Heat itin a boiling water bath. A positive testis indicated by a red color. Mucie Acid Test i Use 1.0% solutions of galactose, glucose and lactose for this test lace 10mL of sugar solution in a small beaker. ‘Ade SmL. concentrated nitric acid and evaporate to approxiriately SmL on he steam bath. Do not ‘evaporate to dryness. ‘Allow the solution to cool, or preferably to stand in your locker until the next laboratory period Note and draw the appearance of any erystal formed under the microscope. Classification of Carbohydrates dently five samples of food found in your home. . List down the carbohydrate present in the ingredients. Classify on what class of carbohydrate does it belong and identify its use. Biochemistry Manual‘Sample Carbohydrate] Classification of Use Present Carbohydrate ‘ Yaka Glucose Monosaccharides | Used to weat hyperkalemia cern ‘Soe Roiownavans TW wast wpeovoet eetay ome tor mone rie |Lactooe, Disoxchiowige, rgoront: savece, Of CnetQy {6° oot ona vo omMONY UiLd in Cod ErEKtui ond prema una Oy eee mat enor Drocaaige Peang: bur Wim ei poe Monaceunariee emote oe Gh ‘sie pe oe, soenonat sg ft On i a e Morescectod eee [Moin come, oF Grek Kor COU ney Mororoccvo.ise eercaiea tose noe nde Nissin OOKe Bertone Roeder [Monasocncnige Twig Ceca PM, Powder Pvocenarige, [Dory Produc rote Papoctoide Yeas Maditin- B. Test for Carbohydrates 1. Write down the significance or purpose for each given test. . 2. Identify the principles involved for each test. 3. Indicate the positive result for each test. Test ‘Significance/Purpose Principle Involved Positive Result Biochemistry ManualALEXIUS CO! i Tciewayneem Todine Test The War GAL Io dittereaote Jouveee Weicot ON. won HRNEOL Pyocmoades (Me OD Used Ho queamiot te premece ot sratcy im vivogieat motels Fodine forms 0 cotgante comer Hoeiweee tee Waco coited yaay rod WKN HE ue doe to jadiorsion THe esr otcsned derceds Juror "HAE OF Te Unioranced or Wiar onaigauatone fot MALE CoMmonoe Armiore -aep bt Fermqpecsin Hecenote cain ord tone canony parle. cdot Gnen- sudden jbrown Wolisch Test A Cremicot tts whic is vied 10 JK For tne presence OF cotwmndory iw 0 aiven Grane Ssimwic ada vederge deaysioier tO Ge neon Gesvotivr- thy compands Jritare with tpha nophiter 10 Sem caarta edicts: rioig Gad Seat wine Woes tia §> [mae mera rer: Crvindory shew Koka si orugtond © 84 VEE Bovoton 6 pedeetd lav the sonction of 0 tomes ‘Osazone Test Reaxng S04, Oion OF ceawitg sey mata ok oR rng adionine route [$c Caghawn. 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