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Emily Cribas, Chem 213W

Isolation and Saponification of Trimyristin to Produce Soap
Introduction
As far back as 3500 BC, humans have been known to use extraction to isolate natural
products from the environment.1 These products have consisted of drugs, perfume, and
cooking spices to name a few. One of these extraction methods includes saponification,
the most commonly used method to make soap. The process was usually accomplished
by boiling animal fat or lard with potassium hydroxide obtained by combining potassium
carbonate from wood ashes with slaked lime (calcium hydroxide) solution.2 For
thousands of years, this process has been used to make one of the most indispensible
tools in societys hygiene belt. Soap is commonly described as an amphipathic fatty acid
salt with both nonpolar and polar properties. The nonpolar nature of soap stems from its
long hydrocarbon chain (the tail). The polar nature is attributed to the charged atom at
the other end of the molecule that defines the soap as a salt (the head). 3 Altogether, this
property allows it to be very efficient at removing unwanted oils, grease, and bacteria.
Specifically, when placed in a dirty environment, the hydrocarbon tails surround
the droplets of grease or other oils, with the polar heads exposed to the water on the
outside. This effectively engulfs the oils in a micelle with the nonpolar ends and trapped
oil on the inside and the head and water on the outside, allowing for the dirt to be washed
away easily with the soap.4 This emulsifying property also makes it effective at washing
away potentially harmful bacteria by forming a micelle around the lipid membrane of
bacterial cells and washing it away with water.

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The purpose of this experiment was to successfully produce soap from triglycerides
isolated from nutmeg using introductory organic chemistry techniques including
extraction and recrystallization. The product from the extraction and purification
(trimyristin) and the product from the synthesis (sodium myristate) were analyzed by thin
layer chromatography (TLC), melting point analysis, and IR spectroscopy. These values
were compared to literature values.
This experiment can be divided into three main steps: 1) isolation of trimyristin from
nutmeg, 2) purification of trimyristin via recrystallization, and 3) synthesis of sodium
myristate via saponification. The scheme outlining the overall process can be seen below
in Figure 1.

Figure 1: Isolation and Saponification Chemical Reaction3

The first step involves the extraction of trimyristin from nutmeg, a common cooking
spice with a history of medicinal use. Nutmeg is composed of 25-40% fats, 8-15%
essential oils, and about 50% cellulose.5 Trimyristin, found in the nutmeg oils, as well as
three other compounds can all be extracted through solid-liquid extraction, due to their
largely nonpolar chemical structure which allows them to be soluble in organic ether
solution (used in the extraction procedure) as explained later.
The second step involves purification of trimyristin. Trimyristin is found in high
concentrations in nutmeg, with no compounds nearing these concentration levels.5 When

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extracted, therefore, trimyristin is found in the highest concentrations with little amount
of other ether compounds. This makes recrystallization the ideal step for purification: it
allows for small amounts of impurities (other compounds) to be left in the solvent layer,
leaving trimyristin to form a pure crystalline structure.
In the final step, saponification, trimyristin undergoes basic hydrolysis after exposure
to an alkaline base to produce soap.3 This process is usually done in the presence of heat,
in this case, via a reflux reaction and is described in further detail below.

Figure 2: Saponification Electron-Pushing Mechanism

There are nine steps in the saponification reaction as seen in Figure 2. The first
step involves nucleophilic attack of the hydroxide (as part of the aqueous sodium

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hydroxide base) on the electron-deficient carbon from the carbonyl bond on trimyristin.
This results in a tetrahedral intermediate where the negatively charged oxygen reaches its
octet by reforming the carbonyl bond, effectively ejecting the rest of the trimyristin fat.
The negatively charged oxygen on the fat now is nucleophilic enough to attach the
hydrogen from the sodium myristate precursor. This results in the formation of the first
sodium myristate molecule and a fat that has replaced one of its ester groups with a
hydroxyl group through hydrolysis. The rest of the mechanism follows these same steps
until there are no more ester groups to hydrolyze. The reaction proceeds because of the
continued presence of the nucleophilic hydroxide in solution and the electrophilic
character of the carbonyl carbons on the trimyristin fat. The entire reaction results in
production of three sodium myristate molecules and one glycerol molecule. These
sodium myristate molecules are fatty acid salts that function as soap.

Experimental
Trimyristin. Ground nutmeg (10 g) was stirred in diethyl ether (30 mL) and refluxed for
30 minutes. The mixture was then cooled to room temperature, and vacuum filtered. The
nutmeg was washed with ether (2 x 20 mL), and the trimyristin solution was evaporated
under a stream of nitrogen to produce a thick, yellow solid. The solid was recrystallized
in warm ethanol (95%, 100 mL). The mixture was vacuum filtered and washed liberally
with chilled ethanol to produce a powdery, white solid (1.14 g, 11.4%). The purity of
trimyristin was checked by TLC (50% ethyl acetate/hexanes). Rf= 0, mp: 54-58C, IR
(ATR) max (cm-1): 2913.5, 2848.7, 1731.8, 1173.1.

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Sodium Myristate. Ethanol (95%, 4 mL) and sodium hydroxide pellets (0.040 g) were
stirred until pellets were dissolved. Trimyristin (200 mg) was added, and the mixture was
refluxed for 20 minutes. The warm mixture was added to distilled water (5 mL) and
saturated sodium chloride solution (10 mL). The solution was stirred and cooled to room
temperature for 10 minutes. The solution was then vacuum filtered with cold water (30
mL) to produce a hard, white solid (2.11 g, 1110% recovery). mp: 98-100C, IR (ATR)
max (cm-1): 3933.12, 2918.93, 2848.09, 1555.30.

Results and Discussion

Natural trimyristin was initially extracted from nutmeg with diethyl ether, along with
other unimportant compounds. It was then purified through recrystallization to get the
pure crystalline solid and eliminate extraneous compounds and impurities. Finally, this
pure trimyristin was refluxed with sodium hydroxide to produce sodium myristate, a
soap. This fun and easy procedure applies several essential organic chemistry concepts
and techniques to help produce a vital hygiene tool from an edible spice. Additional tests
on the sodium myristate were done to understand exactly how soaps work and how it
would react with oils (becomes a homogeneous solution) and ferric chloride (forms a
dark precipitate).
The isolation from nutmeg produced a yellow slushy compound, that when
recrystallized and vacuum filtrated became a completely white, powdery solid. This
extraction worked under the basic concept of diethyl ether (liquid) being able to extract
compounds that are nonpolar (trimyristin) from nutmeg (solid). Trimyristin is extracted
and considered nonpolar because it contains long hydrocarbon chains that make any polar

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groups (carbonyl, ester) it contains sterically inaccessible, allowing it to become miscible
in an organic diethyl ether solvent. The purification worked because there was only a
small amount of other nonpolar compounds and impurities present compared to
trimyristin5, making it fairly easy to recrystallize the main trimyristin solid in a solvent
that is extremely polar and allows for trimyristin solubility only at high temperatures
because of its slight polarity from the carbonyl bond.
The synthesis involved a fairly simple reflux protocol with sodium hydroxide to
produce sodium myristate after exposure to water and salt (all components of soap).
Notably, the varistat was set at 50, and small brown spots were visible in the final
product, indicating that some of it may have been slightly burned.
When compared to the literature values, the isolation and purification steps were
fairly successful at 11.4% recovery, while literature values were at 8%2. Since this is by
mass, a lot of the original mass that was not extracted could pertain to the cellulose and
other major components of nutmeg.
Notably, the percent yield for this reaction was at over 1000%, which is
excessively high when compared to the 80% yield from literature4. This could be
attributed to a couple of reasons. The first, and most obvious, could be that the sodium
myristate may not have dried completely. Since soap is an amphipathic molecule, it is
attracted both to hydrophilic and hydrophobic compounds. This makes it tend to have a
strong attraction for things like water. Another related reason could be that there could
have been residual glycerol from the saponification reaction in the solid.
The IR validates the presence and purity of trimyristin and sodium myristate to an
extent. In the spectrum for trimyristin (Figure 3), there were two distinct aliphatic C-H

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bond stretches at 2913.5 cm-1 and 2848.7 cm-1. Additionally the carbonyl stretch is present
at 1731.8 cm-1. These stretches are not helpful in distinguishing the sodium myristate
from trimyristin as both of them have these stretching frequencies. Most notably, the deep
peak at 1173.1 cm-1 clearly indicates the presence of a C-O ester bond. This piece of
information supports the presence of trimyristin, but does not exclude the idea of sodium
myristate being present. However, melting point analysis of this compound (54-58C)
falls within the expected range (56-58C)7, validating the purity of the isolated
compound. Additionally, TLC results show the expected trimyristin at an Rf of 0.68 when
compared to the ethanol mixture containing of Rf of 0, which supports the idea because
the steric hindrance provided from the bulky hydrocarbon tails of the trimyristin hide the
polar carbonyl group and limit its interactions with the polar stationary phase.
The spectrum for sodium myristate (Figure 4) includes similar aliphatic
frequencies at 2918.93 cm-1 and 2848.09 cm-1, as well as the carbonyl stretch at 1555.30
cm-1. Interestingly, there is a broad peak at 3393.12 cm-1, which is indicative of an O-H
bond. This impurity most likely comes from residual glycerol. An IR with almost
identical values (excluding the OH peak) is also seen in the literature6. This idea is further
validated by the depressed melting point of sodium myristate (98-100 C compared to
330 C).

Conclusions
Overall, the extraction, purification, and reaction were all successful to an extent.
Trimyristin was successfully purified as indicated by IR, melting point analysis, and TLC
comparison. Sodium myristate was most likely synthesized as well, although it was not as

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pure as expected. The presence of sodium myristate is suggested by the IR spectra and
literature comparison. The recovery percentage is a lot higher than expected, but it is not
clear how much of the sodium myristate was actually produced and how much was
glycerol and/or water. If repeated, the refluxing reaction would have been at a lower
temperature to avoid potential burning and more time would have been given for the
sodium myristate to dry and fully reflect the most accurate percent yield.

References
1.

Levey, M., 1959 Chemistry and Chemical Technology in Ancient Mesopotamia,

Elsevier, Amsterdam, The Netherlands.

2. Meszaros, Mark , Russo, T. 1996 Vial Organic, Flinn Scientific Co.,
3. Rummel, S. Experiment 22: Making Soap from Nutmeg, 2011. The Pennsylvania
State University ANGEL Course Fall 2015 Chem 213 Web Site. (Accessed
November 11, 2015)
4. De Mattos, M.C.S.; Nicodem, D.E. Making Soap from Nutmeg: An Integrated
Introductory Organic Chemistry Laboratory Experiment. J. Chem. Educ. [Online]
2002, 79 (1), 9495. (Accessed November 11, 2015)
5. Muchtaridi, Subarnas A, Apriyantono A, Mustarichie R. Identification of
Compounds in the Essential Oil of Nutmeg Seeds (Myristica fragrans Houtt.)
That Inhibit Locomotor Activity in Mice. Inter. J. of Mol. Sci. [Online] 2010,
11(11):4771-4781. (Accessed November 11, 2015)
6. Frank, Forest; Roberts, Theodore, et al. Trimyristin from nutmeg. J. Chem. Educ.
[online] 1971, 48 (4), 255. (Accessed November 11, 2015)
7. AIST: Integrated Spectral Database System of Organic Compounds. (Data was
obtained from the National Institute of Advanced Industrial Science and
Technology (Japan))

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8. Trimyristin, MSDS No. T2500100, Sigma Aldrich www.sigmaaldrich.com
(accessed 12/1/2015)
9. Sodium Myristate, MSDS No. M8005, Sigma Aldrich www.sigmaaldrich.com
(accessed 12/1/2015)
10. Glycerol, MSDS No. G5516, Sigma Aldrich www.sigmaaldrich.com (accessed
12/1/2015)
11. Iron (III) Chloride, MSDS No. 157740, Sigma Aldrich www.sigmaaldrich.com
(accessed 12/1/2015)

Supplemental Information
Spectra

Calculations
1.

% Recovery of Trimyristin=

Dried Trimyristin (g)

1.14 g
100=
100=11.4
Nutmeg (g)
10.0 g

2.

% Yield of Sodium Myristate=

Actual yield (g)

2.11 g
100=
100 = 1110%
Theoretical yield (g)
.189 g