CARBOHYDRATES

1. Molischs Test/A-napthol reaction

Purpose: Detects the presence of Carbohydrates
Reagent: Alpha-napthol and Conc. sulfuric acid
Result: Purple color at the junction.

2. Iodine Reaction
Principle: Iodine forms colored complexes with starch
Results: Red color > Glycogen; Purple color > Starch; Amylose (Dark blue);
Amylopectin (Purple)

3. Benedicts Test
Purpose: Detection of Reducing Sugar (All monosaccharides, lactose, maltose,
cellobiose, and gentiobiose)
Uses: Sodium citrate as sequestering agent, Sodium carbonate, and Copper (II)
Sulfate pentahydrate
Note: Alkali condition converts ketoses to aldoses
Results: Negative is solution remains blue or clear; Yellow, Green, and Red ppt
depending on reducing sugar concentration

4. Barfoeds Test
Purpose: Differentiates monosaccharides from polysaccharides; Ketoses due to
acidity will not isomerise to form aldoses.
Reagent: Curpric ions in acidic medium (Benedicts and Fehlings = basic
solution)
Positive result: Monosaccharides produces a ppt much faster, some
dissacharides will produce ppt but the time difference is significant.

5. Seliwanoffs Test/Resorcinol Test
Purpose: Detects ketones
Reagent: Resorcinol in 6M HCl
Positve result: Rose color (Fructose, honey, hydrolysed inulin)

6. Bials-Orcinol Test
Reagent: Orcinol (5-methylresorcinol) in concentrated HCl with a small amount of
Feric chloride.
Purpose: Test for Pentoses
Results: Pentoses undergo dehydration into fufural that condenses with orcinol to
create a blue-green solution






7. Mucic Acid Test
Reagents: concentrate HNO3 (oxidizing agent)
Purpose: Detection of galactose
Note: Lactose will also hydrolyse to form galactose to yield insoluble dicarboxylic
acid (music acid)
Results: Formation of crystals with sugars or solutions containing galactose

8. Phenyl Hydrazone Test/Osazone Test
Reagents: Phenylhydrazine, Na acetate in acetic acid.
Results: Formation of Osazone crystals (yellow colour) through microscopic
examination.
Glucose and Fructose has same crystals in microscope, but differentiated by
melting point. Sucrose does not form an oxazone.

9. Moores Test
Results: Detection of Monosaccharides
When a solution of reducing sugar is heated with an alkali (NaOH), it turns yellow
to orange and finally dark brown, liberating the odor of caramel. This is due to the
liberation of aldehyde which subsequent polymerizes to form a resinous
substance, caramel. Glucose, fructose or mannose when allowed to stand in the
presence of weak alkali (BaOH) is converted into a mixture containing all the
three sugars and whichever one of them is used the same proportion of the three
sugars is always reached at equilibrium.

10. Fehlings Reagent
Purpose: Detection of reducing sugars (like Benedicts Test and Barfoeds Test)
Reagents:
Fehlings A: Copper (II) Sulfate
Fehlings B: Rochelle salt + NaOH
Use equal volume in testing
Positive Result: Brick Red ppt of Copper (I) oxide

11. Tollens Test
Purpose: Detection of reducing substances (monosaccharides, citrate, and
tartrate)
Reagents: Tollens Reagent (contains ammoniacal silver nitrate)
Result: Silver mirror due to reduction of silver ions into silver metal
Note: This test can differentiate citrate from tartrate because citrate required
boiling to oxidise.








PROTEINS

1. Biuret Test
Use: Detects the presence of peptide bonds/Positive for Histidine
Reagents: 2.5 M NaOH, 0.1 CuSO4
Positive Result: Violet solution

2. Ninhydrin Tests
Use: Detects the presence of amino acids
Reagents: 0.1% Ninhydrin Solution
Positive Result: Polar AA - Blue; Non-Polar AA - Blue VIolet; Proline (imino-acid)
- Yellow

3. Xanthoproteic Tests
Use: Detects the presence of aromatic AA (Tyrosine, Phenyalanine, Tryptophan)
Reagents: conc. HNO3
Positive Result: Yellow ppt, which also turns orange on addition with alkali

4. Millons Test
Use: Specific for TYROSINE
Reagents: 10% Mercury (I) Iodine, NaNO2
Positive Results: Old rose colour (pink-red)

5. Sakaguchi Test
Use: Specific for Arginine
Reagents: A-napthol, 1 gtt of NaBrO
Positive Result: Red colour due to Guanidine group

6. Hopkins Cole Reaction (Glyoxylic acid reaction)
Use: Specific for Tryptophan
Reagents: Hopkins-Cole Reagent, Sulfuric Acid
Positive Result: Violet color (Collagen and gelatin is negative because it does not
contain Tryptophan)

7. Nitroprusside Reaction
Use: Detection of cysteine
Reagents: Sodium nitroprusside in ammoniacal solution.
Positive result: Reddish color (This test is not positive for the sulfur of
methionine)







8. Fohls Test (Lead acetate test)
Use: Specific for sulfur containing amino acids
Reagents: Goulards powder. 30% NaOH
Positive Results: Black ppt due to Lead (II) Sulfate

9. Pauly Diazo Test Test
Use: Detect Histidine, Tyrosine and Tryptophan
Reagents: 1% Sulfanilic acid with NaNO2
Positive Results: Red


Some Functions of Amino Acids

1. Converted to carbohydrates (glucogenic amino acids)
2. Tyrosine forms hormones such as T3, T4, catecholamines, and melanin.
3. Tryptophan is made into Niacin (Vitamin B3) and indole acetic acid (a plant
hormone)
4. Creatine - made of Glycine, Arginine, Methionine
5. Glycine and cysteine - bile salts
6. Glutamate, cysteine and glycine - GSH
7. Histidine > Histamine
8. Glycine is also used for HEME
9. Pyrimidines (Aspartate and Glutamine), Purines (Glycine, Aspartate,
Glutamine, and Serine)
10. Cysteine and Glycine > Detoxicants
11. Methionine > involves in transmethylation reaction
12. Cystine and Methionine > sources of Sulfur
13. Tranexamic acid is a lysine analog
14. Captopril is based on L-Lysine