Chapter 2: Literature Review

2.1 Introduction

In this chapter, the important terms and elements involve and relevant to this research are
discussed. A review about natural sources that is used in this research which is cinnamon
barks. After that, the extraction process, the separation process and the analysis process
are also discussed briefly in this chapter.

2.2 Cinnamon Barks

Cinnamon or scientifically known as Cinnamomum is a popular spice used in

worldwide not only for cooking but also in traditional and modern medicines, where it is
made from the inner bark of trees. As stated by Janick and Jules (2011), the name of
cinnamon comes through the Greek Kinnamomon from Phoenician. While in Indonesia
where cinnamon is cultivated in Java and Sumatra, it is known as Kayu Manis (Samat and
Bell, 2009).

Cinnamon bark is a dried bark of the shoot of evergreen tree that belongs to
Lauraceae family which characterized by oval-shaped leaves, thick bark and a berry fruit.
Mature trees have a thick skin brown or grey and have many branches. Cinnamon is
cultivated by growing the tree for two years, then cutting the stems at ground level. The dried
inner bark is formed strips that curl into rolls which is called as cinnamon sticks. Moreover,
the colour of exterior and interior of cinnamon is light brown and the chemical properties of
cinnamon are spicy, slightly sweet, warm and fragrant.

Figure 2.1. Cinnamon Tree

 There are mainly four types of cinnamon which are Ceylon cinnamon (Cinnamomum
zeylanicum), Indonesia cinnamon (Cinnamomum burmanni), Vietnamese cinnamon
(cinnamomum loureiroi) and Cassia cinnamon (Cinnamomum aromatic). The following table
2.1 shows the main features of the different types of cinnamon.

Table 2.1: main features of the different types of cinnamon (Kawatra and Rajagopalan,
2015)

Cinnamon is often used for medicinal purposes due to its unique properties. For
example, cinnamon bark and leave are widely used as spice and flavouring agents in foods
and various application in medicine. Besides, the essential oil from its bark is rich in trans-
cinnamaldehyde with antimicrobial effect against animal and plant pathogens, food
poisoning and spoilage of bacteria and fungi (Schmidt and others, 2008).

2.3 Chemical composition of cinnamon

Cinnamon consists of various resinous compound such as cinnamaldehyde,

cinnamate, cinnamic acid and numerous essential oil (Senanayake, Lee & Wills, 1978). The
chemical constituents of different part of cinnamon was shown on table 2.2. Singh et al.
reported that the spicy taste and fragrance of cinnamon is due to the presence of
cinnamaldehyde because of the absorption of oxygen. Moreover, the physicochemical
properties of cinnamon were presented in table 2.3.

Table 2.2: Chemical constituents of different parts of cinnamon (Vangalapati et al., 2012)
Part of the plants compound
Leaves Cinnamaldehyde: 1.00 to 5.00%
Eugenol: 70.00 to 95.00%
Bark Cinnamaldehyde: 65.00 to 80.00%
Eugenol: 5.00 to 10.00%
Root bark Camphor: 60.00&
Table 2.3: physicochemical properties of cinnamon (Sangal, 2011).

Parameter Leaf oil Bark oil

Specific gravity (20˚C) 1.030-1.050 1.010-1.030
Optical rotation (˚) (20˚C) 1°96′–0°40′ Slightly laevorotatory
Refractive index (20°C) 1.529-1.537 1.573-1.591
Aldehyde content 4% 65–76%
Eugenol content 77.3-90.5% 4-10%
Solubility characteristics Soluble in 1.5 volumes of Soluble in 2.0–3.0 volumes
70% alcohol of 70% alcohol

In cinnamon essential oil, the main compounds isolated and identified are belongs to
two chemical classes which are polyphenols and volatile phenols. For polyphenols,
cinnamon contains mainly vanillic, caffeic, gallic, protocatechuic, p-coumaric and ferulic acid
(figure 2.2). The chemical compositions of cinnamon essential oils depend on the part of the
plant from that are extracted. In this study, cinnamon essential oil from cinnamon bark was
used. In cinnamon bark essential oil, cinnamaldehyde (figure 2.3) is the most represented
substance depending on the type of extraction. This is being higher for steam distillation than
Soxhlet extraction (Wong & Ahmad, 2014). Another minor volatile compound are
hydrocarbons and oxygenated compounds such as β-caryophyllene, benzyl benzoate,
linalool, eugenyl acetate and cinnamyl acetate (figure 2.3) (Filoche & Soma, 2005).

Figure 2.2. Polyphenolic constituents of cinnamon

 Figure 2.3. Major and minor chemical compounds of cinnamon essential oil

2.4 Essential oil

Essential oils (Eos) are aromatic oily liquids extracted from different parts of plants
that have a collection of hydrophobic secondary metabolites and a volatile natural which has
a strong odour and generally have a lower density than water (Bakkali et. al, 2008). Part of
plant cells that normally produced essential oils are bark, peel and under the surface of
leave, by the helps of the energy from the sun and elements from the air, soil and water.
Moreover, different part of the same plant contains essential oil of different chemical
composition. For example, different part of cinnamon plant possessed varying proportions of
hydrocarbons with primary constituents such as cinnamaldehyde (bark), eugenol (leaf) and
camphor (root), (Gruenwald and Freder, 2010). Thus, cinnamon offers a group of different
oils with diverse characteristic.

Extraction of essential oils become popular and raised scientific interest because of
their biological activity as potential natural agents for food preservation (Velazquez-Nunez
and Avila-Sosa et. Al, 2013). It contains compounds possessing antimicrobial properties,
active against viruses, bacteria and fungi thereby it can be served as natural additives in
foods and food products. Besides, it can be used as active compound in packaging material
because of particularly water vapor barrier property associated with hydrophobicity in nature
of essential oils. As stated by Remmal et al. (1993), essential oils have a very broad
spectrum of inhibition of bacteria including Gram-positive and Gram-negative bacteria. It is
considered as a potential source of natural bioactive molecules and use as alternative for the
treatment of infectious disease (Bouhdid et al., 2006). This is a reason why a thousand years
ago, essential oil has played an important role in our daily lives where it used as anointing
and healing the sick. The first people used essential oils is the Egyptians in medical practice,
beauty treatment, food preparation and in religious ceremony. Apart from that, the Greek
uses the essential oil in therapeutics massage and aromatherapy practices meanwhile the
Romans use the essential oils to promote health and personal hygiene.

Besides, essential oils have a several modes of action as antioxidants. For example,
it can prevent the chain of initiation, free radical scavengers, reducing agent, termination of
peroxides, prevention of hydrogen abstraction from continue as well as quenchers of singlet
oxygen formation and binding transition metal ion catalysts (Yildrim and others, 2000).
Therefore, EOs can serve as the potential natural antioxidant with those functional where it
can be used to prevent lipid oxidation in food systems. This is because phenolics are organic
compounds consisting of hydroxyl group (-OH) attached directly to a carbon atom that is a
part of aromatic ring, so, the hydrogen atom of hydroxyl group can be donated to free
radicals, thereby it prevents other compounds to be oxidized (Nguyen and others, 2003).

2.5 Cinnamon Essential Oil as natural antimicrobial

In this study, the essential oils are obtained from the bark of cinnamon because of its
component is the antibacterial activity against Gram-positive and Gram-negative bacteria
which is responsible for human infectious disease and degradation of food. It is also a
natural preservative that containing active component which is cinnamaldehyde (Ojagh et
al., 2010). In comparative study by Shan, Cai and other members (2005), it shows that
cinnamon has the highest antioxidant activity among 26 spices, so it can be applied as an
antioxidant used in foods. In addition, Cinnamon Essential Oil (CEO) has been found to have
an effective antimicrobial activity and listed as “Generally Recognized as Safe-GRAS” by the
Food and Drug Administration in 21 e-CFR (electronic Code of Federal Regulation) part
182.20 (Valero and Salmeron, 2003).

The cinnamaldehyde has been shown to have broad-spectrum antimicrobial activity

against bacteria, yeasts and moulds (Chen, Wu, McClementsd, Li, and Li, 2017). Several
antimicrobial activities of CEO have been reported in various studies. As stated by Oussalah
et Al. (2006), cinnamon essential oil showed a strong antimicrobial against Pseudomonas
putida strain isolated from meat and Kim et al. (2004) is also reported that the antimicrobial
of cinnamaldehyde is bactericidal against Escherichia coli (E.Coli). According to
experimental that has been done by Tzortzakis (2008), cinnamon essential in the range of
25 and 500ppm had been tested for its activity against certain fungi including Colletotrichum
coccodes, Rhizopus stolonifera, Botrytis cinereal, Cladosporium herbarum and Aspergillus
niger. The in vitro experiment showed that the anti-fungal activity was related to the
concentration of the essential oil. When the 25ppm concentration was test, a 63% reduction
in the production of fungal spore was observed. While at a concentration of 500ppm, fungal
sporulation except B.cinerea was completely retarded.

Apart from that, cinnamon essential oil is also an antiseptic, antibacterial and
anthelmintic properties which make the essential oil is an ideal ingredient to help eliminate
pathogenic bacteria and microorganisms that cause intestinal putrefactions. It is especially in
cases of constipation which food remains longer than necessary in the intestine.
Furthermore, it can treat fungal infections

From the recent studies, cinnamon essential oils can be used as food preservative in
order to improve the shelf life and safety of the food product. Therefore, CEO as natural
bactericide could be an ideal choice to be added into films for the shelf life extension and
quality improvement.

2.6 Overview of Separation Process

The purpose of carrying out separation process is to separate or split the mixture of
the compound into two different individual elements with high purity. This is means that the
individual elements which have been separated have none or very minimum amount of
mixture from others compound. There are a few types of chemical separation which are
distillation, liquid-liquid extraction, solid- liquid extraction and many more. In this research,
solid-liquid extraction process will be conducted to separate the Cinnamon essential oil from
its cinnamon bark.

2.7 Essential Oil Extraction Process

Essential oil is produced when a botanical material is introduced to a solvent in which

some of the plant material components dissolve. So, the solvent becomes infused with the
botanical materials that it has pulled from the source plant. The solution that remains at the
end of the process can be liquid or the liquid can be removed to turn the remnants of the
botanical into a solid. In extraction, function of solvent is to help plant cells to break down
and release their contents. Nowadays, the production and consumption of essential oils are
increasing very fast along with the technology development. Production technology is very
crucial to improve the overall yield and quality of essential oil. There are a lot of extraction
processes to extract essential oil. Example of extraction processes that generally used are
hydro-distillation, steam distillation, solvent extraction, supercritical fluid extraction and
others.
 In this research, Soxhlet extraction method was used to isolate essential oils from
plant material by using food grades solvents like hexane and ethanol. This is because it is
suited for plant materials that yield low amounts of essential oil. Through this process, when
the plant material has been treated with the solvent, it produces a waxy aromatic compound
called a concrete. When this concrete substance is mixed with solvent, the oil particles are
released and then oil is remained in the chemical that used. Usually, this process used in
perfume industry or for aromatherapy purposes.

Based on the technology, people nowadays prefer to use simplest way in extracting
which is Soxhlet extraction. Hence, this method was chosen because it produces high
amount of product. According to Wong and Ahmad (2014), the product extracted with hydro-
distillation was lower than solvent extraction due to the temperature and solvent boiling point
used.

2.7.1 Hydro-distillation Extraction

Hydro-distillation or known as water distillation is a traditional method for removal of
essential oils from aromatic and medical plant. According to Meyer and Warnod et al. (1984),
hydro-distillation is one of the oldest and easiest methods. The main characteristic of this
process is the direct contact between the plant material and boiling water, where the
essential oil is evaporated by heating the mixture followed by the liquefaction of the vapour
in a condenser. The vapour mixture of water and oil is condensed by indirect cooling with
water and then, they are separated through decantation. The function of water in this
process is to boil and carry the essential oil over the condenser along with the steam. There
are three types of hydro-distillation which are water immersion, direct vapor injection and
with water immersion and vapor injection. Times for distillation process is depends on the
plant material. Prolonged distillation will produce only a small amount of essential oil but
does add unwanted high boiling point compounds and oxidation products (Hesham,
Abdurrahman and Rosli, 2016).
Figure 2.4. Hydro-distillation laboratory scale setup
2.7.2 Steam Distillation

Steam distillation is a type of distillation that suitable for a plant that have a
temperature sensitive such as natural aromatic compound. The main point of steam
distillation is to heat and separate the components at temperature below their decomposition
point by adding the water or steam to the distillation apparatus. In this way, many organic
compounds insoluble in water can be purified at a temperature well below the point at which
decomposition occurs. During the process, steam passed through the plant material that
contains the compounds to be separated and then, the steam condensed against the matter
to form a mixture. The system yields a mixture of a vapor and desired essential oil. This
vapor is then condensed further and the essential oil is collected. The solvent used in this
process is water which produced less harmful product and did not cause irritation.
Furthermore, plant part that can be used to produced essential oil by using this method are
berries, ginger, flowers and bark such as cinnamon and sassafras. However, according to
Wong and Ahmad (2014), cinnamon essential oil that extracted with steam distillation was
lower due to some of it stuck to the collection jar during extraction and type of solvent
extraction.

Figure 2.5. Steam distillation laboratory scale setup

2.7.3 Soxhlet Extraction

A Soxhlet extractor is a piece of laboratory apparatus invented in 1879 by Franz von

Soxhlet (Harwood & Laurence M, 1989). It was originally designed for the extraction of a lipid
from a solid material, however, it is not limited to the extraction of lipids but extensively used
to remove water and oil from samples. Typically, Soxhlet extraction is used when desired
compound has a limited solubility in a solvent and the impurity is insoluble in that solvent
(Hesham, Abdurrahman and Rosli, 2016). A simple filtration can be used to separate the
compound from insoluble substance when the desired compound has a significant solubility
in a solvent. Furthermore, it does not require to monitor and manage the operation while
efficiently recycling a small of solvent to dissolve a larger amount of material.

Soxhlet extraction is categorized as traditional method used to isolation of

metabolites from plant material. By using this technique, aroma and quality of oil extracted
from the plant material can be analyte with medium to low volatility (Govendor, 2010). In this
process, the most important is to choose the correct solvent, in order, to obtain a good yield
from the extraction as well as prevent the loss of volatiles. The solvent used in this method is
indicative of the polarity of the compounds extracted.

The principle of Soxhlet extraction is a thimble made from thick filter paper that
contained powdered plant material is placed in the Soxhlet extractor. The extractor which
has a siphoning system is fitted on top of round bottom flask containing extraction solvent.
The Soxhlet is then equipped with a condenser. During the process, the solvent evaporates
and rises to the condenser where it condenses and drains back to the extractor holding the
thimble with the plant material. When the extractor becomes full with hot solvent, the solvent
siphons down to the flask along with the extracted constituents. This cycle may be allowed to
repeat many times over hours or days. During each cycle, a portion of the non-volatile
compound dissolves in the solvent and the desired compound becomes concentrated in the
flask after a few cycles. Usually, after extraction the solvent is removed by using a rotary
evaporator.
 There are a few advantages of Soxhlet extraction which are it is cheap process
because it only requires one batch of solvent to recycled. This method also prevents the
possibility of the solvent becoming saturated with extractable material and enhances the
removal of the analytes from the matrix (Hesham, Abdurrahman and Rosli, 2016). Next, the
temperature of the system is close to the boiling point of the solvent. So, the excessive
energy in form of heat is helps to increase the extraction kinetics of the system. Lastly, the
Soxhlet extraction is give higher yield for certain components or materials to be extracted. It
is proven in the figure 2.6 when comparative studies where carried out by Wong and Ahmad
(2014), it was found that the highest quantity of the extracted oil was Soxhlet extraction by
using ethanol as a solvent.

Figure 2.6. Graph of cinnamon essential oil extraction by steam distillation and
Soxhlet extraction (Wong and Ahmad, 2014).

2.8 Function of starch into edible film

In this research, cassava starch was used to make edible film since it biodegradable
and has good film properties. Starch is classified as polysaccharides which contains the
complex carbohydrates that highly with molecular structure. Most starches are containing
two types of glucose polymer which are amylopectin and amylose. Tharanthan (2003)
reported that the amount of amylose and amylopectin is depend on its source and the
amount of their existing in starch granules also affect the film properties. The importance of
amount amylose in starch is it give the film stronger and high flexibility film due to its linear
chain. Besides, films that are made from starch are describe as odourless, tasteless,
colourless, isotropic and non-toxic biodegradable (Souza et al., 2010).
 Generally, polysaccharides films are very hydrophilic which means that it is poor
water vapor and gas barrier properties. Even though it is not a good water vapor barrier, it
can act as sacrificing agents instead of as a barrier to retard the moisture loss from food
products (Kester and Fennema.1986). This is means that it can relatively be thick films to
intentionally absorb water and provide temporary protection against moisture loss at food
surfaces. Besides, polysaccharides films have a stability structural and ability to slow down
oxygen transmission. According to Banker (1966), film that are not provide protection
against water transmission is usually have a desirable to prevent oxygen transmission and
vice versa. So, film that are made from polysaccharides are very effective as resistance to
gas transmission.

2.9 Effect of glycerol into edible film

Plasticizer is importance in film making especially that used starch as based film.
This is because polysaccharides polymers are hydrophilic with a relatively high degree of
crystallinity. So, it can cause difficulties in processing and performance of film. It also does
not possess like thermoplastic properties. In addition, the problems that usually occurs
during film making is the brittleness and crack on surface of film caused by high
intermolecular forces.

Therefore, glycerol act as plasticizer is added into the film in order to overcome the
brittleness of starch and improving other properties of film. Glycerol is a polyalcohol that
found naturally in animal and vegetables fats and oils. Some authors consider that glycerol is
the best plasticizer for water soluble polymers (Bertuzzi et al., 2007). Souza et al. (2010)
explained that the hydroxyl group present in glycerol are responsible for inter and
intermolecular interaction (hydrogen bonds) in polymeric chains, so, it gives flexible structure
towards film and increase the extensibility of plasticized film.

The interaction between plasticizer and starch is explained by Smits and others
(2003), which the plasticizer interacts with crystalline amylose and amorphous amylopectin
that exist in starch through hydrogen bonding when the temperature is increased and also
during film storage at room temperature. As a consequence, the film will turn like a rubber
due to the reduction of viscosity and increasing of matrix mobility.
2.10 Effect of essential oil into edible film
Essential oil is added to modify the property of edible film properties such as
microstructure, mechanical and barrier. The incorporation of essential oil in forming of
biodegradable film was carried out by applying the emulsifications or homogenization where
fine emulsions of essential oil are obtained contain polymer at aqueous phase.

In recent studies showed by Sanchez-Ginzalez and others (2011), the use of

essential oil can improve the moisture barrier properties of film. This is because essential oil
contains high monoterpenes which hydrophobic substances, so it will increase the
hydrophobicity of films in order to reduce the water vapor migration through film. Besides,
Souza et al. (2013) also reported that homogenous, thin and flexible cassava film were
obtained when added the essential oil. It is also easily to remove from cast. This is because
essential oil is low density, so that it will separate and localized at the upper surface of film.

Apart from that, effectiveness of adding antimicrobial additives directly into food is
low due to the presence of other substances that interact with it, so It will deactivate and
reduce its antimicrobial effect. Therefore, the use of antimicrobial additive in films could be
more efficient since the antimicrobial migrate selectively and gradually from film surface
towards the surface of the food. Then, it can control and prevent from microbial
contamination towards foods.
.