Investigating Fractional Distillation of Organic Fruit Volatiles and Comparison of

Traditional and Microwave Synthesis Routes for n-Butyl Acetate and Benzyl Acetate

August Rothenberger

Department of Chemistry, The Pennsylvania State University, University Park, PA 16802

ajr6121@psu.edu

Abstract ..........................................................................................................................................

The technique of fractional distillation was utilized to separate the components of a liquid mixture of
unknown fruit volatiles. The components of Unknown #3 were identified by boiling point and 1H NMR
spectrum to be ethyl acetate and 1-butanol. Additionally, n-butyl acetate was synthesized using a Dean-
Stark apparatus and characterized by IR, GC, and 1H NMR analyses. Benzyl acetate was synthesized
using a microwave and characterized by IR and 1H NMR analyses. The microwave synthesis route was
determined superior due to its simplicity and efficiency.

Introduction ...................................................................................................................................

Ripening fruits owe their sweet scent to the production of low molecular weight organic volatiles with
pleasant aromas1. These molecules include a wide variety of functionalities, including alkanes, ketones,
esters, and alcohols. Specific concentrations of certain volatiles can be combined to produce the specific
aroma of a particular fruit1. Through the years, researchers have identified many of these molecules and
characterized certain combinations of volatiles so that they might be synthetically produced to provide
artificial scents for a variety of products, including perfumes, soaps, and other cosmetics.

The technique module of this report will involve the separation of two liquid organic volatiles from a
solution using fractional distillation. Distillation can be used to separate liquids by their boiling point
through controlling the temperature of a solution and selectively vaporizing each liquid. However, if
liquids’ boiling points are within about 75°C of each other, the initial azeotropic vapor will contain both
liquids at their molar ratio in solution2. Fractional distillation can solve this problem by allowing the
solution to repeatedly condense and vaporize over a fractioning column. The more condensation-
vaporization cycles that occur, the better the separation of the mixture2. This technique is used to
construct columns with extreme precision to separate complex mixtures and has widespread usage in
large scale petroleum and chemical production industries.

The synthesis module will describe the formation of two esters, n-butyl acetate (1) and benzyl acetate (2).
The key step in both reactions is a Fischer esterification, the formation of an ester from a carboxylic acid
and alcohol under acidic conditions. This esterification mechanism is very common in organic synthesis
due to the widespread usage of the ester functionality. Another esterification mechanism for carboxylic
acids involves treatment with hydroxide followed by reaction with an alkyl halide. Esters can also be
synthesized from acid chlorides or anhydrides in alcohol and pyridine followed by treatment with the
corresponding alkoxide. For the purposes of this report, 1 will be synthesized via esterification of 1-
butanol (3) and acetic acid using a Dean Stark apparatus (Scheme 1). A microwave esterification method
will be used to synthesize 2 from benzyl alcohol (4) and acetic acid (Scheme 2).
Scheme 1. Esterification of n-Butyl Acetate

Scheme 2. Esterification of Benzyl Acetate

Results and Discussion..................................................................................................................

Fractional Distillation of Unknown #3

Unknown #3 was a transparent solution with a dry, vaguely sweet scent known to contain two liquid
organic volatiles. The provided possibilities were pentane, acetone, hexane, ethyl acetate, ethanol, 1-
propanol, water, isobutanol, 1-butanol, and butyl acetate. The fractioning column was loosely packed with
Chore Boy stainless steel wool. The solution was heated until a liquid, Solution 1, was collected at a
maximum temperature of 74.5°C. Once collection of the first product ceased, the column was wrapped in
glass wool and a second product, Solution 2, was collected at a max temperature of 107°C. No fractional
distillation curve could be constructed due to procedural limitations. The products then underwent 1H
NMR analysis on a 400 MHz spectrometer. The annotated spectra are included as supplemental
information.

Using the boiling point and NMR data, Solution 1 can be identified as ethyl acetate (Figure 1). The
boiling point in literature is 77°C, which is consistent with the observed value. Additionally, the NMR
spectrum contains peaks at δ 4.12 (q, 2H), 2.04 (s, 3H), 1.26 (t, 3H), which are the expected peaks and
integration values of ethyl acetate (see Supp. 1). Solution 2 was identified as 1-butanol (Figure 2).
Solution 2’s observed boiling point of 107°C is closer to the literature value for isobutanol (107°C), while
1-butanol boils at 118°C. However, NMR analysis (Supp. 2) showed peaks which corresponded very
closely to those of 1-butanol: δ 3.645 (q, 2H), 1.67 (s, 1H), 1.56 (p, 2H), 1.39 (sextet, 2H), 0.94 (t, 3H).
The 1-butanol spectrum also contains peaks for ethyl acetate. Analysis of the integration shows that
Solution 2 is approximately a 3:1 mixture of 1-butanol to ethyl acetate (Supp. 8-1).

Figure 1. Ethyl Acetate

Figure 2. 1-Butanol
Fractional distillation proved to be a moderately successful method of separating the components of
Unknown #3. While ethyl acetate was effectively isolated from the solution, some remained in solution
and contaminated the 1-butanol sample. This contamination likely resulted in the lowered observed
boiling point of 1-butanol. However, 1H NMR analysis was effective in determining the identities of the
two solutions. The fractional distillation technique leaves much room for improvement. The Variastat
used was not effective at heating the solutions to vaporization, and may have contributed to technique not
achieving complete separation of the ethyl acetate and 1-butanol. Additionally, data collection for
construction of a fractional distillation curve was essentially impossible, requiring the user to count drops
of condensate while simultaneously recording the temperature at regular volume intervals. This method
could not possibly have provided legitimate results for a distillation curve without a degree of data
fabrication. The method could be vastly improved by working with a partner, collecting condensate in a
graduated vial instead, or using a digital thermometer with the ability to record and save data.

Synthesis of n-Butyl Acetate

As displayed in Scheme 1, n-butyl acetate can be synthesized via a single stage Fischer esterification of
acetic acid with 1-butanol. The solution is acidified using Dowex 50 x 2-100 ion exchange resin. The
mechanism for the reaction is activation of the carbonyl followed by attack of butanol and deprotonation
to form a hydrate intermediate. A hydroxyl can be protonated such that reformation of the carbonyl
removes water, then deprotonation of the carbonyl yields the desired product. Since many of the reaction
steps are reversible, the reaction is pushed to completion by using a Dean-Stark apparatus. As the
solution is refluxed, water is preferentially caught in the arm of the apparatus due to its higher density.
Since water is a product, its removal forces reaction equilibrium towards product formation1. After about
40 min refluxing, the product is obtained from the bottom of the apparatus. No work up was performed. A
reaction yield was be determined through gas chromatography.

The product was characterized by IR and 1H NMR spectroscopy. Important peaks in the IR spectrum
(Supp. 3) are the strong C=O stretch at 1738 cm-1 and C–O stretch at 1230 cm-1 which signify formation
of the ester functionality. A 60 MHz NMR was used to analyze the product (Supp. 4); some important
peaks include the triplet at 4.070 ppm of the ester-adjacent methylene, the singlet at 2.046 ppm of the
ester-adjacent methyl, and a vaguely discernable triplet at 1.037 ppm of the terminal methyl. The
remaining methylene protons would ideally split a pentet and sextet, but combine to form a multiplet
around 1.476 ppm due to the low instrument resolution. Additionally, this multiplet integrates to 6 rather
than 4, which is likely due to the analysis software including background peaks in the integration. A gas
chromatogram was taken in CH2Cl2 using a silica stationary phase (Supp. 5). Using the spectrum area
ratios of 1-butanol and n-butyl acetate, the reaction yield of the Fischer esterification was calculated to be
87.5% (Supp. 8-2). This yield assumes that no reactant was lost to the apparatus arm and all reacted 1-
butanol was converted to butyl acetate.

Synthesis of Benzyl Acetate

Benzyl acetate was synthesized via a microwave method using acetic acid and benzyl alcohol. Sulfuric
acid and silica beads were also added. The reaction mechanism is similar to Fisher esterification, however
a microwave is used to bombard the sample with energy through electromagnetic radiation for
approximately 5 minutes. This process is much faster than a using a Dean-Stark apparatus but it requires a
more substantial work up since unreacted reagents and products are left in the same pot. Sodium
bicarbonate was used to neutralize the acetic acid, then an extraction was performed with diethyl ether to
isolate the organic product. The product was then assumed to be pure and a reaction yield was calculated
from the mass to be 69.7% (Supp. 8-3).

Important peaks in the IR spectrum (Supp. 6) are the strong C=O at 1737 and C-O at 1224 cm-1 of the
ester, the aromatic C=C stretching at 1496, 1454, 1380, and 1362 cm-1, and the aromatic out of plane
bending at 735 and 697 cm-1 signifying a monosubstituted aromatic ring. The 60 MHz NMR spectrum
(Supp. 7) is characterized by the singlet at 7.340 ppm of the five aromatic protons, the singlet at 5.100 of
the methylene, and the singlet at 2.085 ppm of the methyl. The aromatic protons do not couple each other
since they are magnetically equivalent.

Synthesis Comparison
Both Dean-Stark and microwave synthesis techniques proved to be an effective method of performing
Fischer esterification. The Dean-Stark method was very tedious and took a longer time, but gave a very
high yield. However, the GC showed that the product was not pure, so the synthesis would likely be
improved by including an extraction work-up step to remove unreacted reagents. Additionally, a mass-
based percent yield would be useful for comparison to the GC yield calculation. The microwave method
allowed for a much more efficient synthesis, both in time required and volume of solvent reacted (24
simultaneous reaction vessels). However, the procedure included a lengthy work-up and showed a lower
percent yield. The microwave method was much more straightforward and polished than Dean-Stark
procedure. For both methods, using a higher MHz NMR spectrometer would have allowed for
confirmation of the product identities with higher certainty, especially for n-butyl acetate. Despite the
lower percent yield, the microwave synthesis should be considered the better method due to its
straightforward and time efficient procedure.

Conclusions
The fractional distillation of Unknown #3 was determined to be moderately successful in separating ethyl
acetate and 1-butanol. 1H NMR was most useful in identifying the products. However, the liquids could
not be completely separated, likely due to equipment limitations, and the procedure could be improved by
implementing a better data collection method. n-Butyl acetate and benzyl acetate were successfully
synthesized at 87.5% and 69.7% yields, respectively, using two different esterification methods. The
products were identified using 1H NMR and IR spectroscopy. Gas chromatography was used to calculate
the yield for n-butyl acetate. Despite the microwave method yielding less product, the procedure was
more time efficient and straightforward than using a Dean-Stark apparatus and it is considered the
superior esterification method.

Experimental .................................................................................................................................

General Methods
Compounds were purchased from Sigma-Aldrich and used without further purification. Reactions were
carried out in open atmosphere. An HP-500 Teflon® Microwave vessel was used for the microwave
synthesis. 1H NMR was performed on a Bruker UltraShield™ 400 MHz and Varian EM360A 60 MHz
NMR spectrometer. Infrared spectroscopy was performed on a Thermo Electron Corporation Nicolet 380
FT-IR. Gas chromatography was performed on a Hewlett Packard HP-6890 Series GC System.

n-Butyl Acetate (1) Dowex 50 x 2-100 ion exchange resin (0.20 g) was filtered then added to a Dean-
Stark apparatus with glacial acetic acid (0.60 g, 34.4 mmol) and 1-isobutanol (0.74 g, 10.0 mmol). The
solution was refluxed for 40 min, then the product was filtered and recovered as a sweet smelling, clear
liquid (87.5%). IR (neat) 1738, 1229; GC (CH2Cl2, silica) RT (min) 5.201; 1H NMR (CDCl3, 60 Hz) δ
4.070 (t, 2H), 2.046 (s, 3H), 1.476 (multiplet, 4H), 1.037 (t, 3H).

Benzyl Acetate (2) Glacial acetic acid (4.0 mL, 70 mmol), benzyl alcohol (2.4 mL, 23 mmol), silica
beads (0.20 g), and conc. sulfuric acid (10 drops) were added to a microwave vessel, which was heated to
120°C over 3 min and held at this temperature for 5 min. The organic product was separated using sodium
bicarbonate and diethyl ether. The organic layers were washed with brine and dried over sodium sulfate to
recover the product as a sweet smelling, yellow liquid (2.41 g, 69.7%). IR (neat) 1737, 1496, 1454, 1380,
1362, 1224, 735, 697; 1H NMR (CDCl3, 60 Hz) δ 7.340 (s, 5H), 5.100 (s, 2H), 2.085 (s, 3H).
References ......................................................................................................................................

1. Masters, K. “CHEM 213M: Fruit Chemistry Module.” Penn State Chemistry Dept. 2018.

2. Williamson, K. L. and K. M. Masters. Macroscale and Microscale Organic Experiments. Cengage

Learning, Boston: 2017.

Acknowledgements .......................................................................................................................

Reagents, materials, spectral instrumentation, and laboratory space were provided by the Penn State
Chemistry Department. Special thanks to Steven Taylor and Prof. Katherine Masters for assistance and
guidance in writing this report.