CARBOHYDRATES

A. GENERAL TEST FOR CARBOHYDRATES

- Liver
- 10 ml water
- 1 ml of 0.1% acetic acid
- Heat 30 minutes (Precipitate the Protiens)
- Filter (Filter paper)
- Collect the filtrate used in Molisch and Iodine Test
1. Molisch Test
- 3 Tubes (glucose, carrageenan, and chicken liver)
- 1st tube
o 2 ml of 10% glucose solution
- 2nd tube
o Pinch of gelatin
o 2ml of hot distilled water
- 3 tube
rd

o 2ml of chicken liver extract

- Add 2 drops of Molisch reagent (Each tube and mix thoroughly)
- 1 ml (15 drops) of Concentrated Sulfuric acid (alongside=create an interface
then record)
2. Iodine Test
- 3 Tubes (glucose, carrageenan, and chicken liver)
- 1st tube
o 1 ml cooked potato starch
- 2nd tube
o Pinch of gelatin
o 2ml of hot distilled water
- 3 tube
rd

- 2ml of chicken liver extract

- Add Drop of iodine solution (Each tube and mix thoroughly)
- Note color
- Warm (60oC)
B. COMPARITIVE TEST REACTIONS OF CARBOHYDRATES
1. Fehling’s Test
- 10 ml vial
- 2 ml Fehling’s A solution
- 2 ml Fehling’s B solution (Note and Record)
- Divide the Fehling’s reagent
- Tubes (glucose, sucrose, fructose, lactose)
- 1 ml of the 10% sugar solution (note any changes)
- Warm (60oC)
- Remove all the tubes when reaction is identified then Record
2. Benedict’s Test
- 7 tubes (glucose, sucrose, fructose, lactose, fasting urine, random urine, and
diabetic urine)
 - Each tube place 15 drops of Benedicts reagent
- 4 tubes with sugar labels 1ml (15 drops) glucose, sucrose, fructose, lactose
- Changes (no change place in warm water bath (60oC))
- Cool at room temp (note changes)
- 3 tube with corresponding urine (fasting urine, random urine, and diabetic
urine) 10 drops of each urine

Report Color of solution Glucose Concentration

(-) Blue < 0.1% or <100 mg/dL
(+) Green 0.5% or 150 mg/dL
(++) Yellow 1.0% or 1,000 mg/dL
(+++) Orange 2.0% or 2,000 mg/dL

C. SPECIFIC TEST FOR CARBOHYDRATES

1. Barfoed’s Test
- 4 tube labeled glucose, sucrose, fructose, lactose
- 1 ml (15 drops) barfoed’s reagent
- 1 ml (15 drops) of test sugar
2. Seliwanoff’s Test
- 3 tube labeled glucose, sucrose, fructose
- 1 ml (15 drops) of Seliwanoff’s reagent
- 1 ml (15 drops) of test sugars
3. Bial’s Orcinol Test
- 3 tube labeled glucose, fructose, xylose
- 1 ml (15 drops) bial’s Orcinol reagent
- 1 ml (15 drops) of test sugar
4. Mucic Acid Test
- 3 tube labeled glucose, lactose, galactose
- 1 ml (15 drops) of test sugar
- 1 ml (15 drops) concentrated nitric acid5
5. Osazone Formation
- 50 ml Vial
o 3 ml of sodium acetate solution
o 2 grams of phenylhydrazine hydrochloride crystals
o Dilute 10 ml distilled water
- 2ml prepared reagent (labelled glucose, fructose, sucrose, galactose)
- 1 ml (15 drops) test sugar