Post-Lab Discussion

Activity 1A

Subcellular Components
of the Living Cell
 Activity 1B

Qualitative Tests for the

Chemical Composition of
Organelles
Tests for Carbohydrates
Molisch Test
Benedict's Test
 Molisch Test

Positive Result: PURPLE ring between

w/ Ribose Sol'n acid and sediment
 Benedict's Test

Positive Result: BRICK RED coloration

w/ Ribose Sol'n
Tests for Proteins

Biuret Test
Xanthoproteic Test
 Biuret Test

Positive Result: PURPLE coloration

w/ Ribose Sol'n
 Xanthoproteic Test

Positive Result: INTENSE YELLOW coloration

w/ Ribose Sol'n
Tests for Lipids

Sudan Test
Acrolein Test
 Sudan Test

Positive Result: RED coloration

w/ Ribose Sol'n
 Acrolein Test

Positive Result: BITING ODOR

w/ Ribose Sol'n
Results
Analysis and Conclusion
 Separation

Principle: Differential Centrifugation

Tissue or cells are first disrupted to
release their internal contents (homogenate).
Larger particles sediment faster than smaller ones
and this provides the basis for obtaining
crude organelle fractions by differential centrifugation.
 Molisch Test
Principle: Dehydration of the carbohydrate by
sulfuric acid to produce an aldehyde

Reaction:
R-CHO + (conc.) H2SO4 →
R-CHO (ex. furfural) + 3 H2O
R-CHO + 2 C10H8-OH (α-naphthol) →
colored product
 Benedict's Test

Principle: Reduction of Cu2+ ions by reducing

sugars, which then can be detected by
the formation of a brick red precipitate
of Cu2O (Cuprous oxide)

General Reaction:
R-CHO + 2Cu2+ + 4OH- →
R-COOH + Cu2O + 2H2O
 Biuret Test

Principle: Reduction of Cu2+ which then

complexes with the N atoms on the
peptide bonds, at high pH.
[Amide bond is resposible for the colour.]

Reaction:
copper(II) ions + protein (peptide bonds) →
biuret complex (PURPLE)
 Xanthoproteic Test

Principle: Nitration of benzene ring present in

the structure

Reaction:
G-H + HNO3 →
G-NO2 (YELLOW) + H2O
 Sudan Test

Principle: Ability of fat cells to selectively

absorb pigments in fat-dyes such as
Sudan IV

Reaction: Absorption
 Acrolein Test

Principle: Whenever fat is heated in the

presence of dehydrating agent, fat
molecule sheds glycerol in the form of
the unsaturated aldehyde - acrolein

Reaction:
Glycerol + KHSO4 →
C3H4O (acrolein w/ biting odor)
END ☺