Scheme of identification of unknown Carbohydrate

If any Change in color: Red-orange-yellow-

Brown : Reducing
1. Molisch test: Give violet ring with all carbohydrates
Glucose – Fructose – maltose - Lactose
2. Fehling test: + 1st boiling
Follow scheme A
Direct flame
If no change in color: Blue – Faint green:

2mls 2mls Non Reducing

Sugar Fehling solution Sucrose – Starch – Gums - Agar

solution
(1ml A + 1ml B) Follow Scheme B
 SCHEME A

Boiling water bath for 2 min. Red ppt: Monosaccharide:

Glucose – Fructose
3. Barfoed test :
No red ppt: Reducing
2mls Concentrated sugar solution + 2mls disaccharide
Barfoed reagent
Maltose - Lactose

4. Osazone test:
Yellow ppt after 15min on hot
Boiling water bath
Monosaccharide: Glucose – Fructose

Yellow ppt after 45min after cooling

0.1gm sugar + 0.3gm Phenyl hydrazine Reducing disaccharide: Maltose – Lactose

5.HCL
Special tests:
+ 0.5gm sodium acetate, Dissolve in
5- Special tests: Check under microscope:
Least amount of water (yellow solution)
If tufts: it is Lactose
If radiating plates: it is Maltose
A- Rapid furfural test:

Sugar crystals+ 3-4 crystals Resorcin + 2mls 12% HCL direct flame deep red color changes to brown ppt Fructose

B- Lead subacetate test:

1ml Sugar solution (distilled)+ 2mls lead subacetate + 1ml conc. ammonia direct flame Faint pink Glucose
 SCHEME B

Off white crystals + SWEET TASTE White + Tasteless Buff (creamy) creamy + brown particles

Sucrose
Starch Gum Tragacanth or agar Gum acacia
Cobalt nitrate test:

Sugar sol. + 3 dps. cobalt nitrate +

xss sodium hydroxide Iodine test:
Permanent violet color 2mls iodine in porcelain dish and sprinkle powders & recognize change in color

Sucrose

Bluish violet particles greenish blue layer wine red particles no change in color

END: Starch END: G-Tragacanth Agar G-Acacia

Stiff gel test oxydase enzyme test

 Stiff gel test: Boil concentrated solution of agar on direct flame then completely cool under tap water Stiff gel

It is Agar
 Oxidase enzyme test: in a porcelain dish moisten the powder with water + 5 dps H2O2 + 5dps Tr. Benzidin deep blue green particles
formed
It is Gum Acacia