 Isolation of organelles in their pure ANALYSIS:

form and analysis of

1. BIURET TEST
macromolecular/biomolecular
- Test for a dipeptide (protein)
components of the organelles.
- Produce pink color from dipeptide to
polypeptide.
PRINCIPLE INVOLVED: Differential
- CuSO4 – Oxidizing Agent ; NaOH – rxn
Centrifugation
medium

STEPS:

1. Extraction
- Process of isolating the organelles.
- Use Saturated Sucrose Solution
(C12H22O11)
- Under mild condition of pH and 2. MOLISCH TEST
temperature under harsh conditions. - General test for carbohydrates.
(T=4°C and <) - conc. H2SO4 – dehydrates a sugar
yielding a furfural.
2. Homogenization – Disrupting the cells - ɑ naphtol in alcohol reacts with
under a mild condition using an furfural to complete a violet/purple
osterizer. colored complex.
3. Centrifugation – Subfractionalization of
the centrifugate.

CENTRIFUGATE/RESIDUE:
I – Nuclear Fraction (Nucleic Acids)
II – Mitochondrial Fraction (Carbohydrates)
III – Microsomal Fraction (Lipids)
3. SUDAN IV
- Classified as oxidizing dye, carcinogen.
- Hydrophobic dye that dissolves on
hydrophobic solvents, lipids.
* Non-specific test because all
hydrophobic will dissolve in Sudan IV
 4. DISCHE REACTION 5. ORCINOL/BIAL TEST
- For deoxypentose - For pentose
- Diphenylamine (DPA) – Putent - conc. HCl – Dehydrates the pentose to
carcinogen. furfural
- Orcinol Rgt. – Gives color reaction with
furfural.

Minced Chicken Liver + 20mL NSS (Normal Saline Solution) [Put to 5 vacuotainers]
1 Residue (Labeled #1) ;
Filtrate 2
Residue (Labeled #2) ;
Filtrate
3
Residue (Labeled #3)
Filtrate (Discarded)

#1: Centrifugated at speed 2 for 10 minutes.

#2: Centrifugated at speed 3 for 15 minutes.
#3. Centrifugated at speed 4 for 20 minutes.

TESTS:

MOLISCH TEST ORCINOL TEST SUDAN IV DPA BIURET

PRECIPITATE (5 Drops) (2 Drops; Water (w/ 2 Drops (w/ 1 Drop conc. ( 2 Drops CuSO4
bath for 5 mins. CHCl3/CH2Cl2) H2SO4 ; Water bath and NaOH)
for 5 mins)
I +++ + + +++ +++
2 ++ ++ ++ ++ +
3 + +++ +++ + ++