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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 (α-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 sol’n?

Y Carbohydrate present

N
Carbohydrate absent

Carbohydrate present
Seliwanoff’s Test (Ketohexoses e.g. Fructose)

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

Bial’s Test (Pentoses e.g. Xylose, Arabinose) 3 mL Bial’s Reagent

Observe color every 1 min for 4mins.
Formation of cherry-red n sol’n?

2 mL test solution Boil Formation of blue-green sol’n?
Y Pentose present N Pentose absent

Y
Ketohexose present

N
Ketohexose absent

Fructose. glucose. maltose 1 mL test solution Barfoed’s Test Monosaccharides 1 mL Barfoed’s 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. xylose. maltose) 1 mL Benedict’s 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 Tollen’s Phloroglucinol Test (Galactose. 5 mL on steam bath (UNDER HOOD) Allow solution to cool Appearance of crystal? Fine. arabinose. Fructose. glucose. yellow Irregular cluster needles (“hay”.Benedict’s Test Reducing sugars (e.) Mucic Acid Test (Galactose) 10 mL test solution in a beaker 5 mL concentrated HNO3 Evaporate to approx.g.g. Cool in water bath Wash w/ distilled water View under LPO Phenylhydrazine Test Reducing sugars (e. Galacturonic Acid) 1 mL test solution 2 mL Conc.of fine needles. xylose. “powde rpuff” like starshaped rodshaped Glucosazone Mannosazone Fructosazone Lactosazone Maltosazone Galactosazone Y Galactose present N Galactose absent . HCl 2-3 gtt Tollen’s Phloroglucinol Reagent Boiling water bath Formation of red sol’n? Y Galactose present N Galactose absent 1 mL phenylhydrazine Mix contents Stopper loosely w/ cotton Heat in boiling water bath Remove after 30 mins. arabinose.

fructose. Observe the volume of the gas evolved at the closed end of the tube. sucrose .Fermentation Test 10 mL 20% Baker’s 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. mannose. glucose. Stopper loosely with cotton. Test for Ethanol 5 mL of the clear filtrate add Lugol’s 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. maltose.g. hexagonal crystals CaCO3 present Iodoform present Presence of D. Take note of the volume of the gas in the closed arm. 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.hexose (e.