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Title: Electrophilic Substitution Reaction Nitration of Bromobenzene and Thin layer

Chromatography

Objective:

The purpose of this lab is to prepare para- bromonitrobenzene from bromobenzene, sulphuric
acid and nitric acid.

MSDS:
 Concentrated Sulphuric Acid
 Corrosive
 Oxidising
 Precautions:
 Wear appropriate Personal
Protection Equipment.
 Avoid eyes and skin contact.
 Locate safety shower and eyewash
station close to chemical handling
area.
 Use EXTREME care when diluting
with water. Always add acid to water
never the reverse.
 People working with this chemical
should be properly trained regarding
its hazards and its safe use.
o Melting Point: -35°C (-31°F)
to 10.36 deg. C (93% to
100% purity)
o Boiling Point: 270°C (518°F)
- 340 deg. C Decomposes at
340 deg. C
o Solubility: Easily soluble in
cold water. Sulphuric is
soluble in water with
liberation of much heat.
Soluble in ethyl alcohol.
 Nitric Acid
 Health hazard/Hazardous to the
ozone layer
 Corrosive
 Serious Health Hazard
 Precautions:
 Keep locked up. Keep container dry.
 Do not ingest. Do not breathe
gas/fumes/ vapour/spray.
 Never add water to this product in
case of insufficient ventilation, wear
suitable respiratory equipment if
ingested, seek medical advice
immediately and show the container
or the label.
 Avoid contact with skin and eyes
keep away from incompatibles such
as reducing agents, combustible
materials, metals, alkalis.
 May corrode metallic surfaces.
 Store in a metallic or coated
fibreboard drum using a strong
polyethylene inner package.
Melting Point: May start to
solidify at -41.6°C (-42.9°F)
based on data for: Nitric acid,
fuming.
Boiling Point: The lowest known
value is 82.6°C (180.7°F) (Nitric
acid, fuming). Weighted average:
98.78°C (209.8°F)
Solubility: Easily soluble in cold
water.
Bromobenzene
 Health hazard/Hazardous to the
ozone layer
 Flammable
 Hazardous to the environment
 Serious health hazard
 Precautions:
 Keep away from heat. Keep away
from sources of ignition.
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 Ground all equipment containing
material.
 Do not ingest. Do not breathe
gas/fumes/ vapour/spray. In case of
insufficient ventilation, wear suitable
respiratory equipment. If ingested,
seek medical advice immediately and
show the container or the label.
 Avoid contact with skin and eyes.
 Avoid all possible sources of ignition
(spark or flame).
o Melting Point: -30.6°C (-
23.1°F)
o Boiling Point: 156.2°C
(313.2°F)
o Solubility: Soluble in
methanol, diethyl ether. Very
slightly soluble in cold water.
 Ethanol
 Skin and eye Irritant
 Precaution:
 Keep away from heat or any source
of ignition
 Do not ingest or inhale gas/fumes
 Wear suitable protective clothing
such as splash goggles
 Ground all equipment containing
material. Do not ingest.
 Avoid contact with skin and eyes
 Keep away from incompatibles such
as oxidizing agents, acids, alkalis,
moisture.
o Melting Point: -114.1°C
o Boiling Point: 78.5°C
Melting Point: -114.1°C
o Solubility: In cold/hot water,
Soluble in methanol, diethyl
ether and acetone.
 Water
 Precautions:
 No specific safety phrase has been
found applicable for this product.
o Melting Point: Not available.
o Boiling Point: 100°C (212°F)
o Solubility: Not Applicable
 Distilled water
 Precautions:
 No specific safety phrase has been
found applicable for this product.
o Melting Point: Not available.
o Boiling Point: 100°C (212°F)
o Solubility: Not Applicable
 Soap solution
 Flammable
 Precautions:
 Flammable liquid and vapour.
 May cause damage to organs.
 Keep away from heat, hot surfaces,
sparks, open flames and other
ignition sources.
o Melting Point: Not available
o Boiling Point: Not available
o Solubility: Not available
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Materials:

1. 2 petridish 7. thermometer
2. 10 ml measuring cylinder 8. stopwatch
3. 2 125ml conical flask 9. funnel with filter paper
4. 1 500ml conical flask for vacuum 10. heating mantle and controller
filtration 11. 2 three fingered clamps
5. hotplate 12. retort stand
6. Organic Distillation Kit 13. beakers

Chemicals
1. 4 ml concentrated Sulphuric Acid
2. 4 ml concentrated nitric acid
3. 3g of bromobenzene
4. hot ethanol
5. cold ethanol
6. cold water
7. distilled water
8. soap solution
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Methods:

Procedure A
1. A 25mL round bottom flask was clamped to a three fingered clamp on a retort stand.
2. A ring clamp was placed under the round bottom flask while on top of the clamp a
beaker was placed with cold water.
3. 4mL of sulphuric acid was measured in a 10mL measuring cylinder as it was carefully
transferred to the round bottom flask.
4. The measuring cylinder was washed as 4mL of concentrated nitric acid was then
measured.
5. The nitric acid was carefully and slowly poured into the round bottom flask that
contained the sulphuric acid.
6. The flask was then unclamped as it was swirled carefully and had cooled in the cold
water bath.
7. The flask was clamped again while over the cold water bath.
8. 3g of bromobenzene was weighed in a clean 10mL measuring cylinder.
9. After a period of 10 minutes was completed the bromobenzene was then added to the
mixture in 0.5mL portions.
10. The contents of the flask were carefully swirled vigorously and frequently after each
addition as it had cooled down in the cold water bath.
11. After the addition was complete and exothermic reaction had gotten subsided a
claisen connecting tube and condenser were then clamped to one end of the connecting tube.
12. A thermometer was placed on the other end of the claisen connecting tube while
making sure that the bulb touched the mixture.
13. A heating mantle and temperature control were connected and was placed under the
round bottom flask as the flask was heated for 15 minutes as the temperature of the reaction
mixture was kept under 60℃.
14. The turning nub of the temperature control was raised to 4.5 port. The heating mantle
was removed when the temperature increased to above 55℃ as it was attached again when
the temperature had been reduced to 40℃.
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15. After the 15 minutes have passed, the mixture was set to cool down slowly to room
temperature as the reaction mixture was then poured carefully with stirring from a 40mL of
cold water in a 100mL beaker.
16. Remnants of the solid from the distillation flask were scrapped as it was transferred
into a beaker.
17. A filtration apparatus was prepared as the filter paper had fit properly on the Buchner
funnel.
18. The Buchner funnel was placed into a 500mL vacuum filtration flask.
19. The rubber tubing was attached at one end to the vacuum filtration flask to the other
end of the vacuum pump.
20. The contents were poured out of the beaker into the Buchner funnel as the pump was
then switched on.
21. The filter cake was thoroughly washed with cold water making sure that the washes
were neutral to the litmus paper (it was confirmed with blue litmus paper).
22. The crystals were drained under the vacuum until it was nearly dry.
23. An empty petri dish was weighed as the solid was transferred into the dish as it was
weighed again.
24. Ethanol was weighed in a measuring cylinder using the following ratio, as for every
gram of crystals was used 1mL ethanol was used.
25. The mixture was heated until the solution had become homogeneous.
26. The solution was cooled slowly to room temperature as it was cooled to 0℃ in the ice
water bath.
27. Another filter paper was prepared in the Buchner funnel to filter the crystals from the
mixture.
28. The crystals were washed with a little ice-cold alcohol (drop wise) allowing the
washes to drain into the filter flask with the mother liquor.
29. After the crystals were dried their weight and melting point were determined. Another
clean empty petri dish was weighed as the crystals were placed on it in order to get the
weight.
30. The mother liquor was transferred to a clean conical flask as it was evaporated to a
volume of about 10mL on a hot plate.
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31. The solutions were then set to cool slowly to room temperature. A second crop of p -
bromonitrobenzene was obtained. The washings were repeated followed by vacuum
filtration.
32. The mother liquor was transferred from the second crop into a clean conical flask as it
was further concentrated by evaporating to a volume of 3-4mL.
33. The mixture was cooled slowly to room temperature. The oil had contained crude o-
bromonitrobenzene.
34. The oil was separated from the 2 phase mixture by means of a micropipette.

Part B Thin Layer Chromatography

1. Solutions were prepared into 2 small test tubes of p-bromonitrobenzene and of the oil
containing o-parabromonitrobenzene in about 0.5mL ethyl acetate.
2. One thin layer chromatography plate was obtained with a pencil as about 1cm was
drawn on a horizontal line as below it p was written for parabromonitrobenzene and o for
ortho-bromonitrobenzene.
3. Spots of each were applied to the two solutions to the sheet as the spots were then set
to be dried.
4. 10mL of the eluting solvent was prepared in a 10mL measuring cylinder with 9:1
hexane: ethyl acetate.
5. After the eluting solvent was ready it was poured into a 100mL beaker as the TLC
plate was placed into the liquid. A watch glass was placed at the top of the beaker as
observations were made.
6. When the solvent was within 0.5cm of the top of the plate the developed
chromatogram was then removed from the chamber as the solvent front was quickly marked
with a pencil (where it had ended) as the plate was set to dry.
7. The spots were visualized by placing the dry plate under the UV lamp, under short
wavelength. Each spot was marked by circling with a soft pencil.
8. The Rf (Retention factor) [distance traveled by substance/distance traveled by
solvent] was calculated for the spots observed as they were identified as either o - or p-
bromonitrobenzene. (A small orange was observed very near to the origin for the oil; this
spot was 1-bromo-2, 4-dinitrobenzene).
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9. The products were stored as they were put into separate lockers to be used for the next
laboratory period.
Column Chromatography
1. A retort stand and burette clamp was used. A 50mL burette was clamped (make sure it
was cleaned) on a burette clamp.
2. A small rolled up piece of cotton was placed into the burette from the top as it was
pushed to the bottom with a long glass rod as the cotton was placed in the narrow part of the
burette.
3. About ½ cm of sand was poured from the top of the burette as it was allowed to settle
on top of the cotton.
4. The solvent mixture (eluent) was poured carefully to about ¼ of the column.
5. 6 g of silica gel and enough solvent mixture was added in a small beaker making sure
that it was all wet (slurry).
6. The silica gel suspension was stirred and carefully poured into the burette, allowing
the silica to settle completely.
7. About ½ cm of sand was poured on top of the settled silica.
8. A beaker was placed at the bottom of the burette as the stopcock was opened allowing
the solvent to drain out just about the level of sand.
9. A teat pipette was used to add 0.2mL of the oil that contained the impure o-
bromonitrobenzene onto the top of the burette.
10. The stopcock was reopened to drain the solvent to let the level of liquid go just below
the level of sand.
11. About 0.5mL of the eluent mixture was added into the column.
12. The stopcock was opened again in order to let some of the solvent drain to let the
level of the liquid be just below the level of sand.
13. The eluent (solvent mixture) was added to the top of the burette.
14. You were now ready to collect. The eluent level was to never go below the level of
sand when collecting fractions.
15. The eluent was collected in 5-8mL fractions in a series of numbered test tubes.
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16. Each fraction was analyzed using the same eluent in the TLC plates by spotting the
fractions on TLC plates. Every 5 fractions were spotted to identify the location of the product
as the adequate fractions were then spotted.
17. The solvent of the fractions that contained the product o-bromonitrobenzene was then
evaporated.

Results and discussion:

1. Report your yield in grams and moles.

o-C6H4BrNO2 p-C6H4BrNO2
Mass (gram) 0.495g 2.744g
Moles (mol) 0.002 moles 0.014 moles
Color (observed and Observed: yellow Observed: very pale yellow
published) Published: yellow Published: colorless to pale
(PubChem) yellow
Melting point (measured) N/A 126.5-130.4oC
Melting point (literature) 40.0- 42.0oC 124.0-126.0oC
(PubChem)

2. Calculate your % yield.

0.495𝑔
% 𝑦𝑖𝑒𝑙𝑑 𝑜𝑓 (𝑜) − 𝑏𝑟𝑜𝑚𝑜𝑛𝑖𝑡𝑟𝑜𝑏𝑒𝑛𝑧𝑒𝑛𝑒 = × 100 = 17.4%
2.847𝑔
2.744𝑔
% 𝑦𝑖𝑒𝑙𝑑 𝑜𝑓 (𝑝) − 𝑏𝑟𝑜𝑚𝑜𝑛𝑖𝑡𝑟𝑜𝑏𝑒𝑛𝑧𝑒𝑛𝑒 = × 100 = 47.5%
5.780𝑔

3. Thin Layer Chromatography report

Mixture o-C6H4BrNO2 p-C6H4BrNO2 Solvent Front

Distance Spot 1: 0.7 Spot 1: 0.8 3.7 4.5cm
Travelled Spot 2: 3.6 Spot 2: 3.6
Rf value Spot 1: 0.2 Spot 1: 0.2 0.8 -------
Spot 2: 0.8 Spot 2: 0.8
𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑙𝑒𝑑 𝑏𝑦 𝑡ℎ𝑒 𝑠𝑢𝑏𝑠𝑡𝑎𝑛𝑐𝑒
(𝑅𝑓 = 𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑡𝑟𝑎𝑣𝑒𝑙𝑙𝑒𝑑 𝑏𝑦 𝑡ℎ𝑒 𝑠𝑜𝑙𝑣𝑒𝑛𝑡 𝑓𝑟𝑜𝑛𝑡)

The TLC results for the mixture came out as expected. Two spots showed up and the Rf

values matched up with the spots created by the (o)-bromonitrobenzene and the (p)-
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bromonitrobenzene Rf values. The TLC results for (o)-bromonitrobenzene did not come out as

expected. The plate showed two spots, one with the Rf value matching (p)-bromonitrobenzene

and the other Rf value was (o)-bromonitrobenzene. This means that the para product was not

completely separated out from the ortho product, the ortho product is impure. The TLC results

for (p)-bromonitrobenzene did come out as expected. The plate showed only one spot and the Rf

value was (p)-bromonitrobenzene. This means that the para product was pure.

Discussion

Table Showing the ortho:para ratio (o:p ratio = weight of oil/weight of p-isomer that
was isolated.)

Ratio p:o = 5.54:1 Grams (g) Volume (mL) % yield (g) % yield
(mL)
(o)- 0.495 0.3 17.4% 17.0%
bromonitrobenzene
(p)- 2.744 1.4 47.5% 47.3%
bromonitrobenzene
The literature value of (p)-bromonitrobenzene is 124.0-126.0oC. (PubChem) The melting

point recorded for (p)-bromonitrobenzene was 126.5-130.4oC. The range for the recorded

melting point has a difference of 3.9oC which is a little broader than the narrow 1-2oC difference

expected for pure melting points. The broader range could have been due to the fibers of the

filter paper that were stuck in the mixed solid and dissolved into the solution. The fibers could

have been in with the separated para product making it impure. Also, it has not been determined

as to why the melting point was higher than the literature value because if the para product was

impure the melting point would have broadened and depressed. Also if it were impure, the

impurity would have shown up on the TLC plate. The melting point for (o)-bromonitrobenzene

was not found, but it would have been expected to be within the 40.0- 42.0oC range. (PubChem)

However, since the TLC showed that (o)-bromonitrobenzene was impure, it could be concluded
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that the melting point would have been broad and depressed.

The data represents the directing influence of the bromide in the bromobenzene through

the products that were synthesized. It was observed than no meta products were formed and that

only ortho and para products were formed (However there was a possibility that 1-bromo-2,4-

dinitrobenzene had formed, but there were no spots observed for the compound on the TLC plate

to prove so). This was expected because of the larger number of resonance structures both the

ortho and para positions have. (p)-bromonitrobenzene was produced as the major product and

(o)-bromonitrobenzene was produced as the minor product which was as expected. The ratio of

the products came out to be 5.54:1. This was not what was expected. In the ratio, more of the

para product and less of the ortho product was produced than expected. Overall, the experiment

resulted in very low yields. It was expected that 85.0% or greater of the expected values would

be yielded, but (p)-bromonitrobenzene only resulted in 47.5% for mass and 47.3% for volume

and (o)-bromonitrobenzene only resulted in 17.4% for mass and 17.0% for volume.

Yields are always less than 100% due to conditions that did not favor complete reaction or to

losses during work-up. Recalling theory as well as observations you made during the

experiment, discuss specific causes of low yield.

The very low yields could have occurred from multiple cases of loss. When synthesizing

the ortho and para products the heat went above 60oC for an extended amount of time. Being

above the 60oC, the 1-bromo-2,4-dinitrobenzene compound could have formed, but like stated

before, it did not show up on the TLC plate so not much of it, if any at all, seemed to form. Since

it went above 60oC, the attempt to cool the round bottom flask down to just under 60oC could

have caused the reaction to not occur, resulting in a lower than expected yield. The thermometer
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during the cooling could have lagged causing the actual heat to be lower than desired, so the

reaction may not have occurred for long. Loss could have also occurred on the spatula, glass rod,

the round bottom flask, the filter flask, and the Buchner funnel. More specifically, a few larger

sized pieces were stuck on the inside of the round bottom flask after the first ice bath. Those

pieces were unreachable and would not wash off with the 95% ethanol. More loss could have

occurred when pieces of the filter paper were removed from the dissolved mixed solid in the

95% ethanol. Some of the liquid could have still been absorbed in the removed filter paper.

To improve the procedure in keeping the temperature at just under 60oC for the entire time

need for the reaction, test with another thermometer to see at what depth in the sand will result in

that ideal temperature. Then when it is time to switch the beaker with the ice and water out for

the sand bath, lower the round bottom flask to that depth and monitor the thermometer, adjusting

the temperature or the depth of the flask in the sand to keep it at a constant temperature just

below 60oC. Also by keeping the temperature constant just below 60oC will help in minimizing

the 1-bromo-2,4-dinitrobenzene that could form. To improve the procedure of decreasing loss on

labware used, empty out the flasks and Buchner funnel as much as possible using the glass rod

and spatula. For the flasks being emptied into the vacuum filtration apparatus, used a pipet and

solvent along with the glass rod and spatula to try and empty them out as much as possible. Swirl

the solvent around to try and get all the pieces attached in places that are unreachable by the

glass rod and spatula. Then make sure to wash of the glass rod and the spatula with solvent or to

wipe them on the dry filter paper if used to get the solid out of the Buchner funnel. To improve

the procedure when removing the dried mixed solid from the filter paper, be careful not to rip off

some of the filter paper with it. If there is filter paper that is ripped off and ends up in the 95%
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ethanol and mixture solution, fish it out with tweezers or a spatula and make sure to squeeze as

much of the liquid out as possible before completely removing it.

Conclusion:

Both the para and ortho bromobenzene was formed. para travel faster than the ortho, which just

stayed in a put position. Ortho was proved to be polar in the eluent system as it was attracted to

the silica in the TLC plate, which is why it stayed still, with little motion. Meta also formed.

Analysis:

1. Explain why p-bromonitrobenzene is less polar than o-bromonitrobenzene?

P-bromonitrobenzene is less polar than o-bromonitrobenzene because it is polarized
in opposite direction with almost the same degree of polarization, this causes
cancellation to occur greatly reducing its polarity. On the other hand,

o-bromonitrobenzene is more polar because it is polarized by two different stru ctures

adjacent to each other acting in the same direction.

2. Write out the resonance structures that contribute to the delocalized structure (σ
complex)
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3. Find the pKas of sulphuric acid and nitric acid in a reference book and confirm that
the latter is the weaker acid. What is the approximate value of the ratio,
Ka(H2SO4)/Ka(HNO3).
𝑝𝐾𝑎 𝐻2 𝑆𝑂4 −3
= = 2.31
𝑝𝐾𝑎 𝐻𝑁𝑂3 −1.3

4. Explain why the melting point of p-bromonitrobenzene is considerably higher than

that of the ortho-isomer.
The melting point of p-bromonitrobenzene is considerably higher than that of

o-bromonitrobenzene because it is more stable, this is because the Br and NO2 are
opposite to each other causing some of the polarity to be cancelled decreasing the
overall polarity contributing to it being stable. More energy is then required to break
the bonds between the p-bromonitrobenzene molecules because it is more stable than
o-bromonitrobenzene.

5. Provide the resonance structure that contribute to the resonance hyrbrids 7 and 8
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a. Use these resonance structures to explain why the formation of m-

bromonitrobenzene is disfavored relative to p-bromonitrobenzene

the carbocation is important in the stability of the product which determines

the product obtained. In the meta isomer the carbocation is located near the
bromine atom which is electron withdrawing making the molecule electrophilic,
this causes two carbons adjacent to each other to be electrophilic and destabilizes
the molecule. Also the carbocation in the para product in not near the bromine
causing the molecule to be more stable than the meta isomer.
6. Explain how TLC may be used to select the most appropriate solvent for use in a
column chromatographic separation?
The solvents for the TLC plate can be changed easily and it is possible to use
several different solvents depending on your desired results. TLC can be used to ensure
purity of a compound. It is very easy to check the purity using a UV-light. Identification
of most compounds can be done simply by checking Rf literature values. You can modify
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the chromatography conditions easily to increase the optimization for resolution of a
specific component.

7. Why does p-bromonitrobenzene have a larger Rf-value in the TLC analysis than does
the o-isomer?
P-bromonitrobenzene has a larger Rf-value because of its polarity being less
than that of o-bromobenzene. This can be deduced from the structure of p-
bromonitrobenzene which is polarized on opposite ends, this causes termination to
occur unlike o-bromonitrobenzene which is polarized by two molecules next to each
other.

8. The o: p ratio in the mononitration of bromobenzene has been reported to be 38:68.

Use this ratio and the amount of p-bromonitrobenzene actually isolated to estimate the
experimental yield of mononitration in the reaction. What errors are there in using
this method to calculate the extent of mononitration?
This is only the total amount of product = 68% and then calculate how much 100%
and find 38% of that and in ratio form 38:68.

Reference:

1. Gilbert, John C., and Stephen F. Martin. "Chapter 15 Electrophilic Aromatic

Substitution." Experimental Organic Chemistry: A Miniscale and Microscale Approach,

Sixth ed., Boston, Cengage Learning, 2016, 2011, pp. 499-522.

2. Hooko, R., et al. "Exp't 52 Nitration of Bromobenzene." Penn State Department of

Chemistry, courses.chem.psu.edu/chem36/New%20Syn%2036%20pdf/Exp52.pdf.

3. McMurry, John. "Chapter 16 Chemistry of Benzene: Electrophilic Aromatic

Substitution." Hybrid, 9th ed., Boston, Cengage Learning, 2016, pp. 478-519.

4. PubChem. National Center for Biotechnology Information, U.S. National Library of

Medicine, pubchem.ncbi.nlm.nih.gov/compound/.