Fisher Esterification Synthesis of Isopentyl Acetate

(Banana oil): picoSpin 45

Introduction
Esters are a class of compounds found widely in nature. Low molecular weight esters tend to
have characteristic flavors and pleasant odors that are most often associated with essential oils,
even though essential oils often are a complex mixture of natural compounds.

The 1R and 2R group side chains can be linear or branch-chain aliphatic or aromatic groups
and they can be the same or dissimilar groups.

The ester functional group can be synthesized by many methods. The simplest approach is
Fisher esterification. By this method, esters are produced by refluxing a carboxylic acid and an
alcohol in the presence of a concentrated acid catalyst. To exploit Le Chatelier’s principle and
shift the position of the equilibrium to the right we add an excessive of one of the reactants to
the reaction mixture. Naturally, we choose the least expensive reactant, which, in this case, is
the carboxylic acid. The reaction mechanism involves initial protonation of the carboxyl
group, nucleophilic attack by the hydroxyl, proton transfer, and loss of water followed by loss
of the catalyzing acid to produce the ester. The process is thermodynamically controlled
yielding the most stable ester product. Typically, only primary and secondary alcohols are used
in the Fisher method since tertiary alcohols are prone to elimination. In this lab, you will
perform a Fisher esterification to synthesize isopentyl acetate from isopentyl alcohol and acetic
acid.

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Fisher Esterification Synthesis of Isopentyl Acetate (Banana oil): picoSpin 45
Purpose

Purpose
The purpose of this experiment is to synthesize isopentyl acetate (3-methylbutyl acetate) via
an esterification reaction between acetic acid and isopentyl alcohol (3-methylbutanol), using
concentrated sulfuric acid as a catalyst. The product will be washed, distilled, then
characterized using NMR spectroscopy.

Literature
Adapted from: Gokel, H. D.; Durst, G. W. Experimental Organic Chemistry; McGraw-Hill,
New York, 1980; pp 344-348.

Weast, Robert C., ed. CRC Handbook of Chemistry and Physics. 70th ed. Boca Raton, FL:
CRC Press, Inc., 1990.

Pulse Sequence
In this experiment, we use a standard 90° single pulse experiment. The recycle delay time (d1)
is adjusted to maximize signal intensity prior to signal averaging the next FID.

Sequence: d1-[0°-aq-d1]ns
0°: Pulse rotation angle (flip angle)
FID: Free induction decay
d1: Recycle delay (μs) for spin-lattice
relaxation
p1: R.F. transmitter pulse length (μs)
aq: Acquisition tim (ms)
ns: # of scans (individual FIDs)

Procedures and Analysis

Time requirements: 3 to 3.5 hours
Difficulty: Moderate
Sample: Acetic acid, isopentyl alcohol, isopentyl acetate

Equipment/materials:

• picoSpin 45 • Iron ring

• NMR processing software (Mnova™) • Ice bath
• Ring stand • Syringe filter (optional filter)

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Procedures and Analysis

• Sand bath (or electric mantle) • Syringe port

• Boiling chips • Port Plug
• Separatory funnel • Tubing
• Thermometer • Reflux distillation apparatus
• Clamps (flask or Keck) • 50 mL round bottom flask
• Simple distillation apparatus • Condenser
• 100 mL round bottom flask • Drying tube
• 25 mL Erlenmeyer flask • picoSpin accessory kit
• Condenser • 1 mL polypropylene syringes
• Three-way adapter • 22 gauge blunt-tip dispensing needles
• Vacuum adapter • Drain tube assembly
• Thermometer adapter • Inlet filter

Physical data

Substance FW (/mol) Quantity MP (°C) BP Density (g/mL)

Acetic acid (anhydr.) 60.05 25 mL 118 1.049
Isopentyl acetate 130.1 product 142 0.876
Isopentyl alcohol 88.15 20 mL 130 0.809
Conc. H2SO4 98.08 5 mL 1.841
5% NaHCO3 84.01 250 mL 1.0018
Sat. NaCl 10 ml
Na2SO4 (anhydr.) 142.04

Reaction

Acetic Acid Isopentyl alcohol Isopentyl acetate

(3-methylbutanol) (3-methylbutyl acetate)

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Safety Precautions

Mechanism

Safety Precautions

NOTICE Be sure that all persons operating this system read the site and safety manual
first.

Experimental Procedure
1. To a 100 mL round bottom flask add 25 mL (0.420 mol) glacial acetic acid followed by
20 mL (0.185 mL) isopentyl alcohol (3-methyl-1-butanol).
Swirl the flask to mix the layers.
2. To the solution add (carefully, gloves) 5 mL concentrated sulfuric acid.
Swirl the flask as sulfuric acid is added (heat generated).
3. Add several boiling chips to the flask, and then place a reflux condenser with lightly
greased joints on the flask as shown in Figure 1.

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Experimental Procedure

Figure 1
Reflux apparatus with exclusion of moisture

Drying tube

Condenser

Water

100 mL round-bottom flask

Sand bath

4. Bring the solution to boil with a sand bath, electric mantel or flame and reflux for 1 hour.
5. After reflux is completed, allow the solution to cool to room temperature.
Transfer the entire solution to a separatory funnel and add 50 mL distilled water. Swirl
the solution, allow the layers to separate, and remove the lower aqueous layer.
6. Add another 25 mL portion of distilled water, shake the flask, and separate and remove
the lower aqueous layer.
7. Extract the organic layer with three 25 mL portions of 5% aqueous sodium bicarbonate
solution to remove excess acetic acid.

Note Be careful, as carbon dioxide is given off during the extraction.

8. Test the last extract and if the aqueous phase is not basic (pH paper), extract the organic
layer with two more 25 mL portions of sodium bicarbonate solution (NaHCO3).
9. After removal of the acetic acid, wash the organic layer with two 5 mL portions of
saturated salt solution.
10. Transfer the organic layer to a 50 mL Erlenmeyer flask and dry over granular anhydrous
sodium sulfate (Na2SO4) or magnesium sulfate (MgSO4).

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Experimental Procedure

11. After drying (the liquid should be clear), decant the organic layer into a 50 mL round
bottom flask.
12. Assemble a simple distillation apparatus, as shown in Figure 2.
Figure 2
Simple distillation apparatus
Thermometer adapter
Three-way adapter
Condenser

Vacuum adapter

Open to air

Receiving flask

Water

13. Add several boiling chips and distill, using a sand bath, electric mantel or flame.
14. Cool the receiver flask in an ice bath.
15. Collect the fraction that distills between 135 °C and 143 °C.
The clear colorless product has an intense odor of bananas; it should be obtained in 80 to
90% yield.

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Analysis

Analysis
Required:
Acquire 1H NMR spectra of:
• Acetic acid (reactant)
• 3-Methyl butanol (reactant)
• 3-Methylbutyl acetate (distilled product)

Optional:

Acquire 1H NMR spectra of:

• Initial reaction mixture (prior to reflux)
• Reaction mixture after reflux
• After H2O wash and NaHCO3 wash
• After NaHCO3 wash, salt extraction and drying

NMR Sample preparation

Using a new, disposable 1mL polypropylene syringe fitted with a 1½ inch 22 gauge blunt-tip
needle draw about 0.5 mL of glacial acetic acid and transfer it to a 0.5 or 1 dram vial.

If available, to this sample, using a new, disposable 1mL polypropylene syringe fitted with a
1½ inch 22 gauge blunt-tip needle, rapidly draw out and add 6 to 10 droplets of
tetramethylsilane (TMS).

Note TMS will begin to boil immediately after insertion of the syringe needle. Thus,
sample transfer must be rapid.

Repeat this procedure for preparing samples of 3-methylbutanol and 3-methylbutyl acetate
for NMR analysis.

Note Some of the optional samples are aqueous and therefore TMS should not be added.

Instrumental Procedure
The general procedure for sample analysis using a picoSpin NMR spectrometer is as follows:

Shim
Ensure the NMR spectrometer is shimmed and ready to accept samples.

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Instrumental Procedure

Presample preparation
1. Displace the shim fluid from the picoSpin capillary cartridge with air.
2. Flush the cartridge with 0.1 mL of chloroform, and then displace the solvent with an air
push.
3. Set up the onePulse script according to parameters listed in the Pulse Script table.

NMR Sample preparation

Using a new, disposable 1mL polypropylene syringe fitted with a 1½ inch 22 gauge blunt-tip
needle draw about 0.5 mL of glacial acetic acid and transfer it to a 0.5 or 1 dram vial.

If available, to this sample, using a new, disposable 1mL polypropylene syringe fitted with a
1½ inch 22 gauge blunt-tip needle, rapidly draw out and add 6-10 droplets of
tetramethylsilane (TMS).

Note TMS will begin to boil immediately after insertion of the syringe needle. Thus,
sample transfer must be rapid.

Repeat this procedure for preparing samples of 3-methylbutanol and 3-methylbutyl acetate
for NMR analysis.

Note Some of the optional samples are aqueous and therefore TMS should not be added.

Injection
1. Using a 1 mL disposable polypropylene syringe fitted with a 1½ inch long, 22 gauge
blunt-tip needle, withdraw a 0.2 mL aliquot of sample.
2. Inject about half the sample.
Ensure all air bubbles have been displaced from the cartridge by examining the drain
tube.
3. Seal both the inlet and outlet ports with PEEK plugs.

Acquire
1. Execute the onePulse script according to the values in the table of parameters provided.
2. Once the onePulse script has finished, prepare the cartridge for the next user by displacing
the sample from the cartridge according to the following protocol: air, solvent, air.

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Processing

Pulse Script: onePulse

Parameter Value
tx frequency (tx) proton Larmor frequency (MHz)
scans (ns) 16 or 25
pulse length (pl) Instrument specific 90° pulse length
acquisition time (aq) 750 ms
T1 recycle delay (d1) 10 s
bandwidth (bw) 4 kHz
post-filter atten. (pfa) 10 (11)a
phase correction (ph) 0 degrees (or any value)
exp. filter (LB) 0 Hz
max plot points 400
max time to plot 250 ms
min freq. to plot -200 Hz
max freq. to plot +1000 Hz
zero filling (zf ) 8192
align-avg. data

live plot

JCAMP avg.

JCAMP ind. Unchecked

a Choose the instrument’s default pfa values

Processing
Download the experimental JCAMP spectra files and open them by importing into Mnova.
The free induction decay (FID) will undergo automatic Fourier transformation and a
spectrum will be displayed.

To each spectrum, apply the following processing steps using the given settings:

Function Value
Zero-filling (zf ) & Linear Predict (LP) 16 k
Forward predict (FP) From aq 16 k
Backward predict (BP) From -2 0

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Results

Function Value
Phase Correction (PH) PH0: Manually adjust
PH1:0
Apodization
Exponential (LB) 0.6 Hz
First Point 0.5
Shift reference (CS) Manually reference
Peak Picking (pp) Manually select peaks
Integration (I) Automatic Selection
Multiplet Analysis (J) -

1. Import each data file into the same workspace in Mnova.

2. Manually apply Ph0 phase correction to each spectrum.
3. Manually shift reference each spectrum using Mnova’s TMS tool.
4. Assign the TMS signal (0 ppm) or CHCl3 signal (7.24 ppm), whichever is present.
5. Identify and assign each signal in the spectra.
6. Save the Mnova document, print each spectrum and paste into your lab notebook.

Results
Acetic acid (Figure 3) contains a carboxylic acid. Proton (1H) NMR spectra of neat carboxylic
acids are identified by a characteristic downfield (high frequency) chemical shift of the acid
proton. The acidic nature of carboxylic acid protons make them strongly deshielded, with
signals typically appearing between 11and 12 ppm. Acidic protons also experience
intramolecular hydrogen bonding and exchange rapidly; rapidly exchanging protons tend to
result in narrow signals. Adding a drop of D2O (heavy water) to the sample causes the peak to
disappear; this is evidence for the presence of a carboxylic acid, but then labile protons from
alcohols, amines, thiols, phenols and enols will also exhibit this exchange behavior. Similarly,
acidic protons experience intermolecular exchange with labile protons from other
compounds, such as water, causing the signal to broaden and shift upfield (low frequency),
closer to the labile proton chemical shift.

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Results

Figure 3
Full 1H NMR (45 MHz) spectrum of acetic acid (anhdyr.; neat)

A characteristic feature of the 1H NMR spectrum of 3-methylbutanol (Figure 4) is the

presence of a coupled alcohol triplet appearing near 5 ppm. One reason a triplet structure is
observed is that alcohol protons undergo dynamic exchange at various rates, fast exchange
leads to hydroxyl protons that do not couple with neighboring protons, resulting in a singlet.
However, slow exchange allows time for coupling, such as that observed in 3-methylbutanol
where the hydroxyl group proton couples with two methylene protons on the neighboring C4
carbon, resulting in a triplet. Steric hindrance and intramolecular bonding can also interfere
with dynamic exchange, allowing for coupling to adjacent protons. The C4 protons (3.5 ppm)
in turn couples with the alcohol and two methylene protons at position C3, generating a
quartet signal instead of a triplet.

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Results

Figure 4

Full 1H NMR (45 MHz) of 3-methylbutanol (neat)

As the reaction proceeds and 3-methylbutyl acetate is produced (Figure 5), two distinctive
spectral features will appear its NMR spectrum. The alcohol signal of 3-methylbutanol will
disappear and the C4 proton quartet will change into a triplet due to the loss of an adjacent
alcohol proton; it will also shift downfield by 0.5 ppm, from 3.5 ppm to 4.0 ppm. The
downfield shift of C4 protons arises from increased deshielding of these protons as the
adjacent alcohol group is transformed into the more electron-withdrawing ester functional
group. Furthermore, evidence of the formation of product is seen in the presence of a singlet
group appearing at just below 2 ppm; this signal belongs to the methyl ester group (C1) of the
carboxyl group (Figure 3 and Figure 5). The remaining signal groups due to protons within the
isobutyl group (C1-3,5 in 3-methylbutanol; C5-7,9 in 3-methylbutyl acetate) are largely
unchanged upon esterification of 3-methylbutanol.

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Comments

Figure 5

Full 1H NMR (45 MHz) of 3-methylbutanol (neat)

Comments
picoSpin 45 1H NMR proton spectra of neat acetic acid, 3-methylbutanol and 3-methylbutyl
acetate are show in Figure 3 to Figure 5. Chemical shifts and related NMR data are available
in Table 1 1H NMR Spectral Data. Chemical shifts are referenced relative to TMS. Spectra are
acquired from neat samples of reactants and product, and aliquots drawn from reaction
mixtures. With the picoSpin NMR spectrometer, it is not necessary to dilute samples prior to
injection. However, 3-methylbutanol is somewhat viscous resulting in broaden signals, more
so than 3-methylbutyl acetate, and dilution to 50% in CDCl3 can improve signal resolution.
Using a labile deuterated NMR solvent will result in the hydroxyl (-OH) protons to exchange
and its signal will diminish or disappear from the spectrum. Likewise, coupling to the
methylene (C4) proton will also be affected.

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Comments

Table 1 1H NMR Spectral Data

Chemical
Figure Compound Signal Group Nuclides Multiplicity
Shift (ppm)
3 Acetic Acid TMS 0 12 H Singlet
HO-C(=O)CH3 2.05 3H Singlet
HO-C(=O)CH3 11.51 1H Singlet
4 3-Methylbutanol TMS 0 Singlet
-CH-(CH3)2 0.90 6H Doublet
-CH2-CH2-CH- 1.47 2H Triplet
-CH-(CH3)2 1.47 1H Multiplet
-CH2-CH-(CH3)2 3.56 2H Quartet
HO-CH2CH2 4.99 1H Triplet
5 3-Methylbutanol TMS 0 12 H Singlet
acetate -CH-(CH3)2 0.91 6H Doublet
-CH2-CH2-CH- 1.53 2H Triplet
-CH2-CH(CH3)2 1H Multiplet
CH3COO- 1.93 3H Singlet
O=CO-CH2-CH2 4.05 2H Triplet

Presented in Figures 6 - 8 are stacked 1H NMR spectra acquired from neat reactants, isolated
product, and spectra acquired from the reaction mixture during various stages of the
experiment. These spectra are instructive in that they demonstrate the need for proper ‘work
up’ of the reaction mixture prior to product distillation and isolation. Moreover, it is easy to
visualize the changes in 1H NMR spectra as reactants are converted to products.

Figure 6 compares reactants and products, while Figure 7 includes the initial reaction mixture
prior to addition of the acid catalyst (H2SO4). We see a hydroxyl signal appearing near
9.3 ppm, the apparent change of the methylene quartet to a triplet (~3.5 ppm), the
appearance of a second methyl ester ‘singlet’ below 2.0 ppm and the broadening of all signals.
The chemicals shift of the –OH group reflects the rapid exchange between the carboxylic acid
proton and the alcohol which coalesce into one signal. Its position is dictated by relative mole
fractions of each component (acid and alcohol); that is, its chemical shift is linearly dependent
on the mole fraction of the two labile protons under exchange. This is a well-known
phenomenon to occur with mixtures of alcohols.

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Comments

Figure 6
Full, stacked and labeled 1H NMR (45 MHz) spectra of acetic acid and 3-methylbutanol (reactants), and
3-methylbutyl acetate (product)

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Comments

Figure 7

Full, stacked and labeled 1H NMR (45 MHz) spectra of acetic acid and 3-methylbutanol (reactants),
3-methylbutyl acetate (product), and the initial reaction mixture prior to reflux.

Looking at Figure 8 we see in the ‘after reflux’ spectrum (spectrum 4th from the bottom)
distinct and resolved singlet resonances due to the each type of carboxyl methyl group; one at
~2.05 ppm from acetic acid and a second one at ~1.93 ppm arising from the methyl ester
product. The methylene ester (-CH2-O-) appears shifted downfield to ~4.0 ppm and is a clear
triplet structure. In addition, due to the presence of mineral acid (H2SO4) the coalesced
hydroxyl/carboxylic acid signal, appearing at ~9.3 ppm in the initial reaction mixture, is
further broaden and is now centered on ~9.7 ppm. The additional downfield shift arises from
an increase of H+ ions to the mixture.

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Comments

Figure 8

Full, stacked and labeled 1H NMR (45 MHz) spectra of acetic acid and 3-methylbutanol (reactants),
3-methylbutyl acetate (product), as well as spectra acquired after different times during the experiment.

After a water wash and neutralization with sodium bicarbonate (NaHCO3), the merged labile
hydroxyl/carboxylic acid proton signal (9.7 ppm) and the carboxylic acid signal (2.05 ppm)
disappear (spectrum 3rd from top). The methylene ester (-CH2-O-) shows better resolution;
also, a residual water signal is present. An additional NaHCO3 wash, followed by salt
extraction and drying of Na2SO4 (or MgSO4) cleans up the product spectrum prior to
distillation (spectrum 2nd from top). The top two spectra in Figure 8 establish that even prior
to distillation, the product 3-methylbutyl acetate is well isolated, and the ‘before and after’
distillation spectra appear nearly identical.

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Own Observations

Own Observations

Ordering Information
To reorder this lesson plan from the picoSpin NMR Spectroscopy: Example Lesson Plans set,
please refer to document order number: “LP52589_E 05/14M picoSpin Lesson Plan
#2-Fisher Esterification Reaction Synthesis”
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