Isolation and Hydrolysis of Trimyristin from Nutmeg

Kerise Hinds
20172044
The Mico University College
Natural Products Chemistry
Facilitator: Mrs. Sophia Haley
November 15, 2021
 Abstract

The objective of this experiment is to extract trimyristin from dried nutmeg and to

hydrolyse the trimyristin to synthesis myristic acid. The trimyristin was extracted from the

nutmeg using the method refluxed condensation where the nutmeg was in reflux with

chloroform for 90 minutes. The chloroform was then distilled under reduced pressure and the

while crystalline solid of the trimyristin were obtain by placing the concentrated solution in

an ice bath. The trimyristin was left to dry for four days in which it was refluxed on a water

bath with 3.5% of ethanolic potassium hydroxide for one hour. The ethanol was also removed

under reduce pressure and the solution was acidified using concentrated hydrochloric acid.

The solid was obtained by cooling the solution in an ice bath.

Various calculations were done based on the mass of the trimyristin and the myristic

acid. The mass of the nutmeg used was 3.5176g. The mass of trimyristin obtain was 0.4172g

and the mass of myristic acid obtain was 0.2545g and 0.4172g of the trimyristin was used to

obtain such mass. The percentage yield was both calculated for the trimyristin and the

myristic acid which gives a percentage of 11.86% and 75.34% respectively. The theoretical

moles of myristic acid were also calculated to be 0.0146moles. The melting point for both

product was unsuccessful.

 Introduction

Natural product chemistry often deals with the isolation of organic compounds from

living organisms. Although natural products chemistry can be conducted on all living things,

it most often takes place on plants. One drawback to these isolation is that they typically very

time consuming and tiresome. This is due to the fact that there are a plethora of compounds in

a plant, so isolating a single one compound will most likely result in a long and complex

process. However, for this experiment the natural product selected was trimyristin. This is a

convenient compound to isolate because it can be found in nutmeg in relatively large

quantises. According to Science direct (2017), “25-40% of trimyristin by weight is found in

nutmeg”.

Trimyristin is extracted from Nutmeg also known as Myristica fragrans Houttayn, is

grown in Indonesia and West Indies. Nutmeg oil is isolated by stream distillation which

consist of approximately 90% of terpene hydrocarbons. Major components are sabinene, and

a- and β-pinene. A major oxygen-containing constituent is terpinen-4-ol. A fraction

consisting of phenols and aromatic ethers, such as safrole, myristicin, and elemicin, is

responsible for the characteristic nutmeg odour.

Trimyristin is an ester with the chemical formula C45H86O6. According to National

Center for Biotechnology information (n.d), “It is a saturated fat which is triglyceride of

myristic acid”. Trimyristin also known as glycerol tritetradecanoate is found in herbs and

spices. It is a constituent of nutmeg. It is a white to yellowish-grey solid that is insoluble in

water, but soluble in organic solvents such as ethanol, benzene and chloroform. Trimyristin

occurs in many vegetable oils as well as myristic acid, it consists of long saturated

hydrocarbon chains and it is relatively non-polar. According to Forrest et al. (1971),

“Trimyristin accounts for the sweet smell in nutmeg as it makes up 20% of dried nutmeg”.
 Trimyristin is classified as a triclyceride, meaning it is a tirester composed of a

glycerol backbone with fatty acid groups that come from myristic acid. To clarify, a fatty acid

group is actually a long chain carboxylic acid. In this case comes from myristic acid.

Trimyristin can be isolated from nutmeg using a relatively non-polar solvent such as

chloroform, by the concept of "like” dissolves “like”.

Myristic acid is a saturated long-chain fatty acid with a 14-carbon backbone which

can naturally be found in palm oil, coconut oil and butter fat. Myristic acid is an oily white

crystalline solid which commonly used in facial cleansers because of its ability to wash away

oils. In addition to that, it helps keeps the skin hydrated and youthful in appearance. The

melting point of myristic acid range from 54-550C. As mentioned before myristic acid can be

synthesis from the hydrolysis of trimyristin and as such the aim of this experiment is to

isolate and hydrolysed trimyristin from nutmeg to synthesis myristic acid.

Image showing detailed mechanism for the preparation of myristic acid from trimyrisin.
Image showing potential undesired (side) reaction that can take place during the hydrolysis of

trimyristin.
 Procedure

Isolation of trimyristin from nutmeg

Approximately 3g of dried nutmeg were weighed and combined with chloroform in a RB

(round bottom) flask and refluxed for 90 minutes. The solution was then distilled under

reduced pressure and filtered obtaining a semisolid that was dissolved in ethanol. With

cooling in an ice bath, trimyristin precipitates were formed, filtered and washed. The filtrate

and washing were kept for the isolation of myristicin.

Saponification

Approximately 0.4172g of trimyristin and 15ml 3.5% ethanolic potassium hydroxide were

refluxed for one hour on a water bath. The flask was allowed to cool to room temperature and

30ml of water were added in which ethanol was removed under reduced pressure. After

filtration the solution were carefully acidified under the fume hood with concentrated

hydrochloric acid and left at room temperature unit the myristic acid solidifies. The beaker

was cooled in an ice bath, the acid were filtered on a Buchner funnel and washed with water.
 Results/Observation

Observation:

The mixture of nutmeg and chloroform while refluxing was pale yellow in colour.

After filtration and the chloroform was removed under pressure a brown solid was obtain.

Ethanol was used to dissolve the solid in which a white cloudy precipitate was formed.

A white crystalline solid was obtain of the trimyristin.

After reflux of the trimyristin and the ethanol was removed, brown soil of the myristic acid

stared to appear. After acidifying the solution and leave it to cool down in a water bath more

of the brown solids started to appear. After drying the solid, appears to light brown powdery

solid.

Results:

Title: TABLE SHOWING THE DATA RECOVERED FROM THE ANALYSIS OF

TRIMYRISTIN OBTAIN FROM THE EXTRACTION OF NUTMEG.

Nutmeg used Trimyristin Trimyristin Myristic Acid

obtained used
Mass (g) 3.5176 0.4172 0.4172 0.2545

Percentage yield - 11.86 - 75.34

%

Molar Mass g/mol - 723.16 - 228.3709

Theoretical moles - - - 0.0146

Number of moles - 0.004864 - 0.0011

Melting point 0C - 56-57 - 54-55

Percent composition of trimyristin in nutmeg: 24-40%

 Calculation

Percentage Yield = _Actual yield____ × 100

Mass used

Percentage yield of Trimyristin = __0.4172_ × 100

3.5176

=11.86%

Number of moles of trimyristin = __Weight of trimyristin utilized__ = ___3.5176___

Molar mass of trymiristin 723.16

= 0.004864moles

Number of moles of myristic acid = ____weight of myristic acid ___

Molar mass of myristic acid
= ____0.2545___
228.3709
= 0.0011 moles

1 mole of trimyristin = 3 moles of myristic acid × moles of trimyristin = 3x moles of myristic

acid.

Theoretical number of moles of myristic acid = 3× 0.004864 = 0.0146moles

Percentage yield of myristic acid =_____Actual mole______ × 100

Theoretical moles

= __0.0011__ × 100
0.0146
= 75.34%
 Discussion

To assess how much trimyristin was effectively extracted and how much myristic acid

was produced by hydrolysis, the percentage yield of trimyristin and myristic acid were

computed. The percentage yield of trimyristin in this example was 11.86 percent, which is

lower than the quantity found in nutmeg, which is typically 24-40 percent. However, this

could have been due to loss of product during the extraction procedure such as spilling of the

solution and transferring from one apparatus to the next. The percentage yield of myristic

acid was calculated by multiplying the moles of trimyristin acid by three to obtain the

theoretical amount of moles that the hydrolysis would yield. In this case, the percentage yield

of myristic acid was 75.34% which was higher than 50%. Nonetheless, this yield could

contain impurities because the product obtain was light brown in colour and myristic acid is a

whit crystalline solid. It is also possible that some side reactions took place which can also

cause not getting 100% yield. If the temperature was kept lower and the product was purified

successfully, it is predicted thar the yield would have been greater.

In order to comment on the purity of trimyristin and myristic acid the melting point

would have to be taken for these products, unfortunately that was not successful. Seeing that

the melting point of trimyristin is 56-570C, it was expected to get a melting point in that range

for the product. But if that was not the case meaning the melting point was lower then

impurities would be present in the sample product. As it relates to the myristic acid it was

evident that impurities were present due to the colour of the crystalline solid that was obtain

which means that the melting point could have been lower or the grange is much grater than

54-55oC. In order to have obtain a pure product recrystallization should have been done but

time did not allow for such activity.

 Conclusion
The theoretical basis and the empirical outcomes were linked in general. Although the

trimyristin % yield was lower than planned, this might be explained by product loss

throughout the operation. Because the sample size is so small, even a small amount of

product lost would have had a significant impact on the % yield. Myristic acid, on the other

hand, was obtained but did not appear to be a pure product. This is not to say that the

theoretical background has no bearing on the results. Trimyristin may be isolated from

nutmeg and refined via recrystallization, according to the findings. Trimyristin can also be

hydrolysed to produce myristic acid. The methods used in this experiment can be used in a

variety of other scenarios. Extraction procedures, for example, are widely used in medicine.

In general, extraction and purification can be used to obtain antioxidant compounds from

Rosemary plants. Despite this, the experiment appears to have accomplished its goal.

Trimyristin was extracted, and the resulting product was hydrolysed to give myristic acid.
 References
Forrest, F., Roberts, Snell. T, etal. (1971.). Trimyristin from nutmeg. Journal of Chemical

Education. 48 (4): 255. . doi:10.1021/ed048p255.

https://en.wikipedia.org/wiki/trimyristin. Accessed October 27, 2021

McCullough, P.; Guy, W. Melting Point Purity Determinations: Limitations as Evidenced by

Calorimetric Studies in the Melting Region. Analytica Chimica Acta 1957, 17(1), 80-

96. https://jcmarot.files.wordpress.com/2021/06/lab-report.pdf. Accessed October 27,

2021.

National Center for Biotechnology information (n.d). Trimyristin. Pubchem database.

https://pubchem.ncbi.nlm.nih.gov/compound/trimyristin. Accessed October 27, 2021

Science Direct. (2017). Trimyristin. https://www.sciencedirect.com/topics/agricultural-

biological-sciences/trimyristin. Accessed October 27, 2021.

Raw Data from the Experiment
Image showing trimyristin in the round bottom flask after was placed in the ice bath.
Image showing solid myristic acid.