Mata, Ritchelle Abigail P.

2009-50507

Unknown Determination: Carbohydrates

Aug. 16, 2012 PH CH 127.1

TEST FOR POLYSACCHARIDES (starch, glycogen, gum arabic) 5 mL test solution Drop iodine solution Color change? Y Polysaccharide present (Intense blue solution) N Polysaccharide absent (Clear, colorless solution)

TEST FOR CARBOHYDRATES (starch, glucose, sucrose and other sugars) Molisch Test 0.5 mL test solution 2 mL water 2 drops Molisch Reagent (-naphthol + H2SO4) Mix well; incline test tube 3 mL conc. H2SO4 (slowly!) *pour on side of test tube Purple layer at junction? Y N Carbohydrate absent

Anthrone Test 1 drop solution 0.5 mL water 1 mL Anthrone Reagent Swirl carefully Boil for water bath (3mins) Cool and observe

Formation of blue-green soln?

Y Carbohydrate present

N
Carbohydrate absent

Carbohydrate present
Seliwanoffs Test (Ketohexoses e.g. Fructose)

5 mL Seliwanoff Reagent
1 mL test solution Boil in water bath (1min.)

Bials Test (Pentoses e.g. Xylose, Arabinose) 3 mL Bials Reagent

Observe color every 1 min for 4mins.

Formation of cherry-red n soln?

2 mL test solution Boil Formation of blue-green soln?

Y Pentose present N Pentose absent

Y
Ketohexose present

N
Ketohexose absent

Benedicts Test Reducing sugars (e.g. Fructose, glucose, xylose, arabinose, maltose) 1 mL Benedicts Reagent 2 gtt test solution Mix well Boil in water bath (3mins) Allow solution to cool down Formation of brick-red precipitate? Y Presence of reducing sugar N Absence of reducing sugar

Tollens Phloroglucinol Test (Galactose, Galacturonic Acid) 1 mL test solution 2 mL Conc. HCl 2-3 gtt Tollens Phloroglucinol Reagent Boiling water bath Formation of red soln? Y Galactose present N Galactose absent 1 mL phenylhydrazine Mix contents Stopper loosely w/ cotton Heat in boiling water bath Remove after 30 mins. Cool in water bath Wash w/ distilled water View under LPO

Phenylhydrazine Test Reducing sugars (e.g. Fructose, glucose, xylose, arabinose, maltose 1 mL test solution

Barfoeds Test Monosaccharides 1 mL Barfoeds Reagent 2 gtt test solution Boil in water bath for few minutes Allow solution to cool down Formation of brick-red precipitate after 2 mins Y Presence of monosaccharide N Absence of monosaccharide
*Disaccharide present if ppt formed after several (15 mins.)

Mucic Acid Test (Galactose) 10 mL test solution in a beaker

5 mL concentrated HNO3
Evaporate to approx. 5 mL on steam bath (UNDER HOOD) Allow solution to cool Appearance of crystal?

Fine, yellow Irregular cluster needles (hay- of fine needles, powde rpuff like

starshaped

rodshaped

Glucosazone Mannosazone Fructosazone

Lactosazone Maltosazone Galactosazone

Y Galactose present

N Galactose absent

Fermentation Test
10 mL 20% Bakers yeast suspension in Eihorn Saccharometer 4 mL test solution
Invert tube several times until well mixed Fill the closed end of the tube (avoid air bubbles) Do not fill the open arm (mouth) to give for water displacement. Stopper loosely with cotton. Observe the volume of the gas evolved at the closed end of the tube.

5 mL gas produced?
add 5 mL of 40% sodium hydroxide
fill open arm w/ water Cover the mouth using thumb and shake *make sure all the gas is in the closed end of the tube before removing thumb. Take note of the volume of the gas in the closed arm.

Test for Ethanol

5 mL of the clear filtrate add Lugols solution *until a yellow color is produced

Test for Carbon Dioxide

2-3 mL of the clear filtrate

2 gtt CaCl2 TS
White precipitate

Foul odor, bright yellow precipitate, hexagonal crystals CaCO3 present Iodoform present

Presence of D- hexose (e.g. glucose, fructose, mannose, maltose, sucrose