Name: Ahmad Abdulkadir Arabi

American University of Nigeria

DEPARTMENT OF NATURAL AND ENVIRONMENTAL SCIENCE

INTRODUCTION TO BIOLOGY

BIO120 LAB 7

SEPARATING ORGANIC
COMPOUNDS
Abstract:
This experiment is primarily concerned with paper chromatography.
Among these three items are column chromatography, paper
chromatography, and gel electrophoresis, all of which are used to
separate organic molecules from mixtures.
Introduction:
Separation procedures are required for the separation of various types of
materials. The emphasis in this lab is on paper chromatography and gel
electrophoresis and how they function. In this lab session, paper
chromatography is used to separate the various chemical compounds in a
sample and the pace at which they disperse. The stationary phase (the
paper fibers) and the mobile phase are the two primary phases in paper
chromatography (this is the organic solvent that moves along the paper).
It is a simple approach for separating various chemical compounds. Gel
electrophoresis is a technique for separating molecules based on their
charge, shape, and size (morgan and carter). When this happens, an
electric charge is delivered through the gel, causing it to migrate from
one end to the other. The resulting Rf value (retardation factor)
characterizes a known molecule in a known solvent under known
conditions, and is calculated as follows:
Distance moved by sample
Rf = Distance ¿
origin ¿ solvent front ¿

Materials:
o Chromatography paper
o Mortar and Pestle
o Gloves
o Pencil
 o Leaves
o Carrots
o Sand
o Knife
o Chromatography solvent
o Jar
o Acetone
o Pipette
o Test tube
Methods:
Procedure 7.1: The submitted samples (lemongrass and carrot) were
mashed with sand. The colors were then extracted using acetone. A tiny
beaker was filled with the extracted liquid. Then, from either side of the
chromatography paper, a 1cm line was drawn. The extract was then
placed in a pipette and gently extracted along the line on the paper. A
solution of 30ml was put to a beaker. After that, the paper was folded
and inserted with the line facing the liquid. The paper was eventually
withdrawn. Observations were made and then documented.
Results:
The line on the paper began to detach and migrate up the paper away
from the liquid. The deeper green line at the bottom of the page faded
and lightened. It changed from a dark green to a light green color. Below
is the graphic description of the experiment.
 CHROMATOGRAPHY DATA FOR DETERMINING AMINO ACID UNKNOWNS

Amino Acid or Distance to Solvent Distance Traveled by Rf

Sample Number Front Sample
1 12 cm 6.5 cm 0.54 cm
2 12 cm 7 cm 0.58 cm
3 12 cm 8.7 cm 0.72 cm
4 12 cm 9 cm 0.75 cm
5 12 cm 11.6 cm 0.97 cm

Conclusion:
The chromatogram demonstrated how each structure of molecules move
s at a different rate and appears at a different position on the finished chr
omatogram. In conclusion, paper chromatography is an effective method 
for separating organic compounds in a mixture that cannot be separated 
using other techniques.
Experiment 2;
Materials
o Pipette
o Electrophoresis chamber
o Agarose
o 4 dyes (methylene blue, xylene cyanol, bromine and orange G)
Procedure 7.3: This experiment dealt with separating organic
compounds using process of Gel electrophoresis, it started off by the gel
been created. The mold has two open ends, therefore must be taped
tightly and repetitively. After pouring the agarose liquid into the mold, it
was mandatory for a ‘comb’ being placed in the mold to create the wells
as the liquid solidifies. After 20 minutes, it solidified, the comb and tap
were removed and the gel was placed in the chamber. The buffer
solution is used to deliver the current to the agarose gel. the buffer
solution is poured to the brim so it covers the gel. Addition of one of
each sample of the dyes to separate wells using a pipette. The chamber
was covered and made sure the negative side of the circuit is on the
same side as the wells. After two hours of sitting in the electricity, one
of the samples (dye) was visible and started moving centimeters away
from the starting well and so the others started moving too.
Results:
Below is a picture of before and after Gel electrophoresis

Fig 1: Fig 2:
In Fig 1, the samples were in a gel bed before underdoing
electrophoresis. And in Fig 2, one of the samples started moving while
in electricity.
Conclusion:
In conclusion, gel electrophoresis is a procedure that, while potentially 
dangerous, is an excellent instrument for determining the pace at which 
molecules travel based on their electrical charge and total size.
References:
Morgan, Judith Giles and M. Eloise Brown Carter. Investigating Biology: Laboratory Manual 8th Edition.
Harlow UK: Pearson, 2015.