INSTITUTO TECNOLÓGICO Y DE ESTUDIOS SUPERIORES DE MONTERREY

Obtaining Nanomaterials, Organic and Bioinorganic Materials

Group 303

Module 3

Dr. Saeed Beigiboroujeni

Collaborative Activity : Delivery of Final Project Report

Group 3 members:

María Jolie Cabrera Maldonado A00833129

Mauricio Rojas Ley A01721821

Pedro Cortés García Luna A01721754

Paulina Aragón Flores A01284649

December 1st 2022

Name of the product to be synthesized: Noxidin (Edible antioxidant coating for

fruits and vegetables)

Common name of raw material: Sesame seeds → Sesame Oil

Names and license plates of the members, day of the laboratory and group:

Group 3

María Jolie Cabrera Maldonado - A00833129

Mauricio Rojas Ley - A01721821

Pedro Cortés García Luna - A01721754

Paulina Aragón Flores - A01284649

November 25th 2022

Theoretical Framework

Importance of Raw Material Use

Due to the final purpose of our product proposal (edible coating for fresh cuts of
organic materials) we need a material that has a compatibility with the ingestion of
the human diet. Thus leads the investigation to a material that brings health benefits
through ingesta.

Currently, sesame seeds (Sesamum Indicum) are used in the food industry as a
supplement for obtaining and processing the human body; as they contain
phytosterols, antioxidants, vitamins, calcium and proteins.

Its high lipid content gives the opportunity to take advantage of sesame oil
derivatives; which has high percentages of oleic acid, linoleic acid, palmitic acid and
stearic acid. This derivative has applications in the cosmetology industry, as it is a
product that can be easily organically incorporated into the manufacture of
shampoos, conditioners, make-up bases and moisturizing lotions. (Gómez, 2019)

Sesame seed (Sesamum Indicum) production is relatively sustainable, there is no

known significant damage to air, water, land, soil, forests. It takes 9,371 liters of
water to produce 1 kilogram of sesame seeds.

Sesame seed oil is also very versatile in terms of business opportunities, mainly
because it is used in many areas such as food, cosmetics and dietary supplements.
This makes sesame oil very good product to be sold as a product of its own, it can
be used by many people in different products such as cosmetics like cream or it
could be used as an ingredient in cooking recipes, making it ideal for starting a
business.

Ingredient description and properties

Sesame seeds (Sesamum Indicum) are made up of 86 phenolic compounds, 26 non

phenolic compounds (Mekky, 2019), phytosterols (a structure very similar to
cholesterol with recognized cholesterol-lowering effects), mucilages (a type of
soluble fiber that lowers cholesterol, triglycerides and blood glucose levels) and
flavonoids (flavones (15), flavonols (8), flavanones (2), and a flavan-3-ol).(Morsy,
2022)

The seed is small, from 2 to 4 mm, flattened in shape, with a flattened shape, of
variable color between creamy white and black.

More than half of the weight of the seed is oil, and the rest: protein (18%), fiber (8%),
minerals (2%), etc. The fats it contains are unsaturated, with a predominance of
polyunsaturated fats. These, together with the also present lecithin, make sesame a
food that contributes to reducing blood cholesterol levels.

The protein of these seeds is made up of fifteen different amino acids, with a high
proportion of the essential amino acid methionine. On the other hand, sesame seeds
have good amounts of fiber, so that their consumption is beneficial for the regulation
of the function of the digestive system. beneficial for the regulation of intestinal
function.

Description of the product to be developed

The product is an antioxidant film cream to delay oxidation in fruits and vegetables.
The product is called Noxidin and the sesame oil properties aid to maintain the
freshness.It is made by non harm ingredients such as sesame oil, citral, polysorbate
20, sodium alginate, glycerol and water. Technically edible, however, it has not been
tested whether there’s any secondary effects after consumption. It presents a citric
smell and has a thick and slimy consistency. It is an opaque color.
Relevance of the nanostructure to be used in the product

Making the product into a nanoemulsion has advantages over just having a normal
emulsion. A nanoemulsion provides more stability to the colloid at a low cost of
production. Also they have a higher carrying capacity which helps the effectivity of
the product antioxidant function.

The nanoemulsion is also relevant because of the homogeneity of the particles,

meaning that when the coating is applied to the fruit of choice, the coating will act
evenly in all parts of the fruit where it was applied. This means that the fruit will stay
anti-oxidized evenly and will not have parts where the fruit stays fresh for less time
than other parts.

Need covered by the product

The product seeks to address SDG #2 zero hunger and #13 climate action. Because
it would reduce the amount of fruit that goes to waste and help feed people who do
not have the resources to buy more food. Also, the green production of this product
contributes to the creation of innovative projects practicing green chemistry.

Observations on the experimental procedures

Sesame Oil Extraction

● Ethanol is more efficient than isopropyl alcohol
● Using a heating mantle provides a consistent and uniform heat throughout the flask
● Using a cotton cup is very recommended as it expels more oil than the filter paper
bag
● Covering the lower part connector with aluminum foil to prevent the evaporation of
the solvent.
● Controlling the lipidic product as it can denaturalize if it’s let too much under the heat.
Extraction purification

● Rotavapor at 60ºC was used to separate the oil from the solvent.
● Using a decantation funnel to separate impurities and heavier components not
needed.
Raw Material

● Grinding the seeds expel more oil than non-grinded.

● A cotton cup works is very efficient as a filter.
 Image 1. Use of rotavapor to separate
the solvent from the sesame oil at the end
of the Soxhlet extraction
Image 2. Decanting of oil extraction; the separation seen is of compounds from the sesame
seeds that were washed out during Soxhlet extraction.

Sintesis Characterization Procedures

Acetyl Chloride Test

● Test for detecting alcohols in an organic sample.

● If there is a presence of alcohols, ammonium chloride gas will present as white
fumes.

Ambient pressure
and

temperature values (1 atm at 25 °C)

 Image 3. Acetyl Chloride test on sample. We can appreciate the smoke being a product of
the reaction with ammonium hydroxide; this phenomenon indicates the presence of alcohols
in our organic compound.

Lucas Test

● To detect primary alcohol, secondary and tertiary alcohols.

● In this case, primary alcohol is reported in the oil.

Ambient pressure and temperature values (1 atm at 25 °C)

 Image 4. Lucas Test on sample. We can appreciate that the lucas reagent solution did not
react as a precipitate when added to the sample action indicating the presence of primary
alcohols (OH present in linoleic and oleic acid).

Ferric Chloride Test

● Test for presence of phenols

● If the sample turns blue, purple, red or green it is positive. In the sesame oil, phenols
are present according to the test.

Ambient pressure and temperature values (1 atm at 25 °C)

 Image 5. Ferric Chloride Test on sample. We can appreciate the green color that was
formed homogeneously in the solution; an action that indicates the presence of phenols.

Litmus Orange Paper Test

● Litmus orange, obtaining an acidic result.

Ambient pressure and temperature values (1 atm at 25 °C)

 Image 6. First pH test

Image 7. Second pH test. Upper one is the tested with oil, lower one is the paper without
anything

While analyzing the pH of the sesame oil extracted, a 6-7 pH was observed which can be
contrasted to the 5.5 pH of the regular sesame oil, however it is theorized that the pH is not
close to the standar pH due to the use of ethanol as a solvent which could’ve left some
residue thus affecting the pH.

Thin Layer Chromatography

● Retentions factors were calculated after the TLC.

To determine the retention factor it is necessary to measure from the baseline to the solvent
front, all this is the distance traveled by the solvent.
This will be the X. This is equal to 3.3 cm.

The oil separated into 2 components.

The measurement from the baseline to the spot of analyte which is the last point the oil
reaches, all this is the distance traveled by the solute.This will be the Y.

The first mark is 1.6 cm, which is the first component of the separation of the sesame oil.
To calculate the retention factor it is needed to divide the Y over the X.

Y
Rf=
X
1.6 cm
Rf=
3.3 cm
R f =0.485

The second mark is 2cm. The second component of the separation.

 Y
Rf=
X
2 cm
Rf=
3.3 cm
R f =0.606

Ambient pressure and temperature values (1 atm at 25 °C)

 Image 8. Thin Layer Chromatography of sesame oil under UV light. 50/50 hexane and
ethanol. We can appreciate the difference of compounds in silica gel paper by identifying
their paths.

Product Formulation
Image 9. Nanoemulsion after ultrasonication. A homogeneous substance of soft color can
be seen, which indicates that the emulsion was made in the correct way.

Image 10. Formulation of the edible coating for the final product. The image shows how
sodium alginate is added to the glycerol contained in a beaker.
Image 11. Nanoemulsion is added to the edible coating of sodium alginate with glycerol in a
dropwise manner.

Results of each stage

Oil extraction characterization

Color Through extraction it took on a dark

brown/yellow color

Smell It is almost odorless, but has a typical

sesame note and smells slightly spicy,
toasted and greasy

Texture Smooth texture with oily complexion

Flavor Toasted and intense, like nuts

Chart 1. Sesame oil organoleptic specifications

Running Weight (g) Performance Density pH Smoking

sample after % Point
decantation (lipids)

1 6.47 16% 0.840 6.5 177 °C

2 6.45 16% 0.837 6.5 172 °C

3 6.50 16% 0.842 6.5 173 °C

 Chart 2. Sesame oil physical specifications

Functional Component Chemical/ Result

Groups/Phytochemi Names Phytochemical
cals Test

Quercetin 3-O- Acetyl Chloride Positive

rhamnopyranosi Test
de
Alcohol Components
Myricetin Acetyl Chloride Positive
Test

Luteolin C- Acetyl Chloride Positive

deoxyhexoside Test
C-hexoside III

Luteolin Acetyl Chloride Positive

Test

Linoleic Acid Lucas Test Positive

Primary Alcohol
Oleic Acid Lucas Test Positive

Sesamol Lucas Test Positive

Sesamol Ferric Chloride Positive

Phenolic Compound Test

Sesaminol Ferric Chloride Positive

Test

Vanillic Acid Ferric Chloride Positive

Test

Chart 3. Chemical/Phytochemical composition of sesame oil obtained experimentally

After the final product was done, two different analyses were performed in the
nanoemulsion and in the extraction. The first one was the Infra-red spectroscopy,
which was used to determine the functional groups of both the extraction and final
emulsion and the last analysis was DLS which was made to determine the average
particle size in the final nanoemulsion and product.
 Graph 1. Raw material’s Infrared Spectroscopy analysis

Analyzing the graph of the raw material, there is the signature OH slope at around
the 3300 line, which is part of some compounds found in sesame oil such as
sesamol and sesaminol. There is also the carbon-hydrogen stretch at the 2800 mark
which is true for molecules like sesamol, sesaminol, and oleic acid which are all
organic compounds made by carbon and hydrogen and in some cases alcohols
which were also displayed in the IR spectra. After the 1800 line we can see the
footprint of the sesame oil.

Product Characterization

Graph 2. Nanoemulsion Infrared Spectroscopy analysis

This graph shows that the major functional group that is present in most of the
organic molecules that make up sesame oil is the alcohol group represented by the
slope that is located around the 3300 line. After that there is really not a functional
group distribution in the IR spectrum that is really noticeable, but compared to a
research made by Anand Prakash, et. al. The nanoemulsion IR spectra that they did
also had the same peaks of the IR spectrum that we obtained in the process of the
nanoemulsion.

Graph 3. IR spectra of citral, sesame oil and tween 80 nanoemulsion (Prakash, A, et,
al., 2019)

Based on this spectra comparison we can see that the nanoemulsion as our final
product resulted as expected, with the different functional groups and with the IR
footprint that it should have based on a previous investigation where they did the
same nanoemulsion, just changing the tween 20 with tween 80 which was the one
they used in the research paper.

Graph 4. Nanoemulsion Dynamic Light Scattering DLS analysis

Analyzing the graph, it can be seen that the sizes that are most abundant in the
nanoemulsion are in the range from 10 to 100 nm in size. There are some particles
sizes which are less abundant but have a larger range, varying from 200 to 650 nm,
this can be because of the ultrasonication that was just made once for 2 minutes and
15 seconds, to reduce the size of those particles one thing that could be change is
the amount of times the sample is sonicated and the time of each sonication in order
to have a less dispersed and more concentrated particle sized. There were also
some small molecules in the range from 1 nm to 10 nm, this is a good sign that the
sonication process was successful and if donde multiple times could give a
nanoemulsion with more particles in those sizes.

Ingredient Volume Density Weight (g) Total w/w %

(mL) (g/mL) weight (g)

Sesame Oil 0.0178 0.92 0.0164 0.0157

Citral 0.0892 0.888 0.0792 0.0757

Tween 20 0.107 1.095 0.117 0.112

Water 101.785 1.00 101.785 104.598 97.31

Sodium N/A N/A 2 1.912

alginate

Glicerol N/A N/A 0.6 0.5736

Chart 4. Concentrations of all the ingredients used in the preparation of the product
elaboration

Product quality attribute Scale

1 = nonexistent
9 = very intense

Characteristic and expected color 9

according to the type of product A thick, white-colored product was
expected to be obtained.

Characteristic and expected smell 9

according to the type of product Due to the citric aldehyde in the
formulation, a fresh odor was expected
to be obtained.

Weird odors 1
Unexpected odors were not obtained

Strange particles 1
The substance was homogeneous
throughout its structure.
 General appearance according to 6
declared flavor The product has no flavor, however, due
to the strong citral odor a rare
experience can be identified.

Chart 5. Quality attributes and scale used in product sensory evaluation

Feature Room Temperature Refrigeration

Color and odor Thick solution of whitish Thick solution of whitish

color and citric odor color and citric odor

Physical state: solid, Liquid Liquid

liquid, resin, etc.

Homogeneous or Homogeneous Homogeneous

heterogeneous mixture

Hygroscopic No No

Lumps or fungi No lumps or fungi No lumps or fungi

detected detected

Chart 6. Product properties at different temperatures

How to use the product: Once the package is opened, place it in a cool place at
room temperature (22 °C - 27°C) to avoid contamination. Depending on the size of
the food, place from 2 to 3 drops on the exposed surface, spread with a kitchen
utensil to cover as much as possible and wait for the coating to dry. If the food will be
consumed in approximately 2 to 3 days, place it in the refrigerator. If it is consumed
the same day, leave it in a cool place.

Nutrition Facts
Serving size 80 ml
Calories: 5.15

Total Fat 0.0166 g 22%

Saturated Fat 0.0023 g 10%

Trans Fat 0 g 0%

Polyunsaturated Fat 0.0071 g

Monounsaturated Fat 0.0071 g

Cholesterol 0 g 0%

Sodium 160 mg 7%
 Total Carbohydrates 1 g 1%

Fiber 1 g 4%

Sugars 0 g 0%

Proteins 0 mg 0%

Humidity 4.6 %

Ashes 6.54–7.71%

Chart 7. Nutritional information regarding the amounts of reagents in our final product

Results discussion
Different tests were performed in order to characterize the extraction
obtained. Such tests were the acetyl chloride test for the presence of alcohols, the
Lucas test for the distinction of alcohol types, ferric chloride test for the presence of
phenols and a purity test with TLC. All of the tests point towards the expected
compound from the extraction, sesamol. Errors could’ve been made because there
wasn’t too much oil to use for characterization, using small quantities of it could’ve
meant there wasn’t much room for an evident. There was also the possibility of
ethanol still being present in the extraction.

The acetyl chloride test showed the presence of alcohols which corresponds
to the compounds found in sesame oil. As mentioned before there is still the
possibility of this reaction occurring because of the ethanol. The Lucas test showed
the presence of primary alcohols. Sesamol and sesaminol consists of phenols which
would not react with the Lucas reagent. This showed that there are no compounds
with secondary and tertiary alcohols, which holds true to the compounds within
sesame oil which mostly consists of carboxylic acids. Nonetheless this does not help
us prove or disprove the presence of ethanol in the extraction. The ferric chloride test
showed the presence of phenols which holds true to sesamol and sesaminol
compounds.

The purity test was done with TLC in which two different compounds were
separated because of the different retention factors. Since sesame oil has many
different compounds, more tests were done with the pure compounds of oleic acid
and linoleic acid in order to find the relative retention factors. We were not able to
obtain sesamol and sesaminol in its pure state to obtain the relative retention factors
of the separate compounds to compare them with the TLC made from the extraction.
By analyzing the properties such as molecular weight, polarity and structure, it was
determined that the two compounds found in the TLC of the extraction were sesamol
and sesaminol respectively.

A litmus paper test was performed to determine the presence of phenols due
to them being slightly acidic. Orange litmus paper was used in order to measure the
pH of the extraction solution (sesame oil), if the paper turned slightly yellow it would
indicate a change of pH to the acid side. When the drops of sesame oil were added
to the litmus paper, the color changed to a slight yellow, showing a positive result in
the presence of phenols in the extraction. Phenols in the extraction are due to the
compounds sesamol and sesaminol, and the test proved that the extraction of
sesame oil contained these two substances. It is important to have these substances
because sesamol and sesaminol give the extract some antioxidant properties used
for the final product of the nanoemulsion.

In regards to the IR spectra of the extraction of the sesame oil, the most
notable groups were the OH valley and the C-H stretch, which relates to the main
compounds in sesame oil which are linoleic and oleic acid, sesamol and sesaminol,
that are all organic compounds with carboxylic acids and alcohols along with carbon
chains. The IR spectra of the nanoemulsion coincided with the one in the research
by Anand Prakash, et. al. which showed the same major OH valley and the same
footprint at the end of the IR spectra.

The DLS analysis proved that the emulsion was in the nanoscale. This is
because most of the particles detected by the DLS were in the range of 10 to 100 nm
in size. Most of the particles were between 10 nm and 100 nm, but other particles
appeared in the smaller range from 1 to 10 nm and a some particles were in the
ranges from 100 nm to 1000 nm because of uneven sonication or because the
sample was left to set and some particles conglomerated and formed bigger
molecules. Overall, the nanoemulsion had some dispersion but a very small
distribution in the nanoscale.

Conclusion
In conclusion, based on the chemical tests done to the extraction of sesame
oil, it was determined the extraction was successful because presence of primary
alcohols and phenols were present in the extract due to a positive result in the acetyl
chloride test, no precipitation in the lucas test and the green color from the ferric
chloride test. Finally, the TLC was used to ensure that the extract contained sesamol
and sesaminol. These tests proved the presence of sesamol and sesaminol, two
compounds that offer some antioxidant properties to the extraction and thus useful in
the final product of the nanoemulsion to help protect the fruit from oxidation. The
DLS analysis helped in determining the distribution of sizes of nanoparticles and the
IR spectra in finding the most common functional groups in both the extraction and
the final nanoemulsion product.
The compounds analyzed in the characterization were useful to the final
product because they contribute antioxidant properties for the protection of the fruit it
were applied on, as well as helped in making the emulsion completely edible with the
same properties that other solutions could have but with a higher toxicity that would
make the coating dangerous for human consumption.

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