BIOCHEMISTRY LAB REPORT 6 2023| PORTAGE
LEARNING|WITH PROCEDURES, EXPLANATION AND FINAL
RESULTS.HIGHLY VERIFIRD
Column Chromatography and TLC of Spinach
Group Experiment 3 – Individual Lab Report
(Save as pdf and submit by NOON, 1 week after your experiment)
Comments for Grading TA:
(Please indicate if you performed the lab on a day other than your regularly scheduled day
and/or with a TA other than your regular TA).
Page Limit: report must not exceed FIVE pages (including this page)LIMIT DOES NOT INCLUDE ANY
GRAPHS, SPECTRA, OR REFERENCES
(Please see General Lab Report Guidelines for detailed descriptions of all other requirements)
DEDUCTIONS FOR HANDWRITTEN REPORT/STRUCTURES/GRAPHS
AND EXCEEDING PAGE LIMIT
LATE PENALTY IS 2 MARKS PER DAY LATE (NOT ACCEPTED AFTER 7 DAYS)
, Chemistry 212 FALL 2022
Group Experiment 3: Column Chromatography – Lab Report Guidelines
Include the following in your report (in the following order, use provided table):
Objective: (3 mark)*
What is the purpose of this experiment?
The purpose of this experiment is to extract chlorophyll a and b as well as beta-carotene
from the spinach leaves. Then, column chromatography is performed for separating the
crude extract. Thin Layer chromatography(TLC) is proceed to analyze the different effects
of various solvents which is helpful when it comes to choosing an appropriate solvent for
maximum separation on the column.
Introduction: (3 marks)*
Brief description of the concept/reaction studied – extraction, column chromatography –
and why it is important
Extraction is a useful technique that is able to remove unwanted reagent from targeted
product. It is suitable to use when the polarity of wanted and unwanted compounds is
varied dramatically. In this experiment, extraction is used to obtain the pigments from
spinach leaves. However, extraction is not able to separate the three kinds of pigments of
experiment. Thus, column chromatography is necessary.
Column chromatography is used to purify compounds based on their polarity. All the
compounds are put in the column of silica gel which is a highly polar substrate. Non-polar
solvent (eluent) will carry the compounds run forward through the column. This makes
that molecules with less polarity will run faster than the more polar one thus the
compounds in the mixture are able to be separated.
These two techniques (extraction and column chromatography) are both very useful in the
chemistry lab which make them very important.
Procedure: (3 marks)*
Summary of lab procedure – what YOUR GROUP did, include KEY observations – be
concise! Someone should be able to repeat your experiment from this summary alone.
Preparing Column: Measure 20 ml silica gel and 150 ml eluent and prepare a 30 cm
column. Push a small piece of cotton to the top of the bottom and clamp it in a vertical
position. Then, cover the cotton with a thin layer of sand and tap the column gently to level
the sand. Add about 8ml eluent into the column. On the other hand, mix the gel with 35ml
eluent in 250ml flask which will give a mixture called “slurry”. Swirl the flask. Once the
sand settled, poor the slurry into the column. Open the stopcock to drain eluent into 250ml
Erlenmeyer flask. When the eluent is 6cm above the silica, add 1cm sand to the top of the
silica. Continue to drain until the eluent is level with the top of the sand. Close the stop
cock.
Extraction: Take 3-4 leaves and shred them into a mortar. Add sand with 8ml methanol in
it. Crush with a pestle for a minute. Methanol will gradually turn light green during this
process. Decant the liquid through a funnel into 125ml separatory funnel. Add 5ml
methanol and 15ml petroleum ether to leaves and repeat the process before (Grind the
leaves and decant through a funnel into separatory funnel). The solution should be green
after the procedure. Repeat the process before with 7ml methanol and 15ml petroleum
ether. The solution should be dark green. There should be two layers formed inside
2 of 5
LEARNING|WITH PROCEDURES, EXPLANATION AND FINAL
RESULTS.HIGHLY VERIFIRD
Column Chromatography and TLC of Spinach
Group Experiment 3 – Individual Lab Report
(Save as pdf and submit by NOON, 1 week after your experiment)
Comments for Grading TA:
(Please indicate if you performed the lab on a day other than your regularly scheduled day
and/or with a TA other than your regular TA).
Page Limit: report must not exceed FIVE pages (including this page)LIMIT DOES NOT INCLUDE ANY
GRAPHS, SPECTRA, OR REFERENCES
(Please see General Lab Report Guidelines for detailed descriptions of all other requirements)
DEDUCTIONS FOR HANDWRITTEN REPORT/STRUCTURES/GRAPHS
AND EXCEEDING PAGE LIMIT
LATE PENALTY IS 2 MARKS PER DAY LATE (NOT ACCEPTED AFTER 7 DAYS)
, Chemistry 212 FALL 2022
Group Experiment 3: Column Chromatography – Lab Report Guidelines
Include the following in your report (in the following order, use provided table):
Objective: (3 mark)*
What is the purpose of this experiment?
The purpose of this experiment is to extract chlorophyll a and b as well as beta-carotene
from the spinach leaves. Then, column chromatography is performed for separating the
crude extract. Thin Layer chromatography(TLC) is proceed to analyze the different effects
of various solvents which is helpful when it comes to choosing an appropriate solvent for
maximum separation on the column.
Introduction: (3 marks)*
Brief description of the concept/reaction studied – extraction, column chromatography –
and why it is important
Extraction is a useful technique that is able to remove unwanted reagent from targeted
product. It is suitable to use when the polarity of wanted and unwanted compounds is
varied dramatically. In this experiment, extraction is used to obtain the pigments from
spinach leaves. However, extraction is not able to separate the three kinds of pigments of
experiment. Thus, column chromatography is necessary.
Column chromatography is used to purify compounds based on their polarity. All the
compounds are put in the column of silica gel which is a highly polar substrate. Non-polar
solvent (eluent) will carry the compounds run forward through the column. This makes
that molecules with less polarity will run faster than the more polar one thus the
compounds in the mixture are able to be separated.
These two techniques (extraction and column chromatography) are both very useful in the
chemistry lab which make them very important.
Procedure: (3 marks)*
Summary of lab procedure – what YOUR GROUP did, include KEY observations – be
concise! Someone should be able to repeat your experiment from this summary alone.
Preparing Column: Measure 20 ml silica gel and 150 ml eluent and prepare a 30 cm
column. Push a small piece of cotton to the top of the bottom and clamp it in a vertical
position. Then, cover the cotton with a thin layer of sand and tap the column gently to level
the sand. Add about 8ml eluent into the column. On the other hand, mix the gel with 35ml
eluent in 250ml flask which will give a mixture called “slurry”. Swirl the flask. Once the
sand settled, poor the slurry into the column. Open the stopcock to drain eluent into 250ml
Erlenmeyer flask. When the eluent is 6cm above the silica, add 1cm sand to the top of the
silica. Continue to drain until the eluent is level with the top of the sand. Close the stop
cock.
Extraction: Take 3-4 leaves and shred them into a mortar. Add sand with 8ml methanol in
it. Crush with a pestle for a minute. Methanol will gradually turn light green during this
process. Decant the liquid through a funnel into 125ml separatory funnel. Add 5ml
methanol and 15ml petroleum ether to leaves and repeat the process before (Grind the
leaves and decant through a funnel into separatory funnel). The solution should be green
after the procedure. Repeat the process before with 7ml methanol and 15ml petroleum
ether. The solution should be dark green. There should be two layers formed inside
2 of 5
