Chitin and Chitosan

Chitin comes second as the most abundant polysaccharide in nature after cellulose. It is naturally
present in the exoskeleton of crustaceans (such as crab, lobster, shrimp etc.) and various insects,
worms, fungi, and mushroom in varying amount. Chitin chemical structure, poly-β-(1→ 4)-N-
acetyl-D-glucosamine, is similar to cellulose besides in chitosan one hydroxyl group on each
monomer has substituted with an acetylamine group (ref 1., ref 2.). Chitosan, natural carbohydrate
polymer derived by chitin deacetylation, has been widely utilized as natural food preservatives
proven for their antimicrobial and antifungal properties against a range of food-borne
microorganism (ref 1.). Kanatt et al. (2007) whom observed chitosan-mint (CM) mixture
effectiveness towards meat and meat products showed that the shelf life was enhanced in chilled
CM treated samples. Besides, chitosan based preservatives has also been proven to be effective in
enhancing the shelf life of bread, egg, fruit and vegetables, juice, kimchi, mayonnaise, meat, milk,
noodle, rice cake, seafood and seafood product, soybean curd (tofu), and vinegar (no2007).
Moreover, shrimp-derived chitosan has also been submitted to US Food and Drug Administration
(FDA) to be considered as GRAS (generally recognized as safe) based on scientific procedures for
multiple technical effects.
In order to be used as food preservatives, chitin must firstly be extracted from chitin-rich source
and be further processed to chitosan. So far, the main commercial sources of chitin are various
crustaceans, principally crab and shrimp shells (younes2015). Chitin isolation from shellfish
requires removal of two major constituents of the shell, proteins by deproteinization and inorganic
calcium carbonate by demineralization, together with small amounts of pigments and lipids that
are generally removed during those two steps (younes2015). A wide range of chemicals have been
tested as deproteinization reagents including NaOH, Na2CO3, KOH, K2CO3, Ca(OH)2, Na2SO3,
NaHSO3, CaHSO3, Na3PO4, and Na2S. NaOH is the preferential reagent and it is applied at
concentration ranging from 0.125 to 5.0 M, at varying temperature and treatment durations
(younes2015). Demineralization, the process of mineral removals, is mostly performed using
dilute hydrochloric acid (younes2015).
Chitin obtained from previous process is later converted to chitosan by deacetylation. During this
process, acetyl groups are removed but also depolymerization reaction occurs, indicated by
changes in chitosan’s molecular weight (younes2015). This process can be achieved by enzymatic
preparations or chemical process (younes2015 masukin ref di papernya). Chemical methods are
more commonly used for commercial purpose of chitosan preparation because of their low cost
and suitability to mass production (younes2015 masukin ref di papernya). From a chemical point
of view, either acids or alkalis can be used in this process. However, glycosidic bonds are very
susceptible to acid; therefore, alkali deacetylation is preferred (younes2015 masukin ref di
papernya). Commonly, hot concentrated solution of NaOH is used in deacetylation process,
resulting an insoluble chitosan residue deacetylated up to ~85%-~99% (younes2015 masukin ref
di papernya).
Orange peel essential oil
Essential oils (EOs) are volatile substances in a form of liquid extracted from different parts of
medicinal and aromatic plant (laranjo2017). Naturally, EOs functions as a plant’s protective agents
for their antimicrobial, antiparasitic, insecticidal, antiviral, antifungal, and antioxidant properties.
In food industry, EOs are widely used as antioxidants added to food preservatives (laranjo2017).
The application of thyme/cinnamon EOs, rosemary/thyme EOs, and oregano EO are proven
effective in increasing the shelf-life of meat and meat products (laranjo2017). The antimicrobial
effect of various EOs on food products vary according to the composition of each food products,
the characteristics of each food product, and the specific microorganisms to be eliminated
(laranjo2017). Citrus essential oils appear as interesting natural compounds with great potential
use in foodstuffs preservation (Laura 10.1016). Viuda-Martos et al. (2008) evaluated the effect of
lemon, mandarin, and orange oils on the growth of moulds associated with food spoilage and found
out that orange and mandarin essential oils were the most effective against Aspergillus niger and
Aspergillus flavus, respectively. Frassinetti et al. (2011) whom evaluated the antimicrobial and
antioxidant activites in orange, lemon, and mandarin showed that those family of citrus exhibit
antimicrobial activity at 15-250 µg/mL MIC and antioxidant as 20-70% DPPH.
One type of citrus which is most commonly found in Indonesia is orange. Orange is the second
largest fruit commodities produced in Indonesia, around 2.2 millions tons produced anually in
2016. Orange essential oil are obtained from orange peel extraction by conventional method such
as simple distillation, solvent extraction, maceration, cold pressing, and modern techniques like
effleurage, Super Critical CO2 extraction, and Turbo Distillation Extraction. Modern methods are
costly and laborious. Macreation is not preferable because it changes the composition of oil also
solvent extraction leaves traces of solvent in the extract which may cause allergies and affect
immune system. Therefore, steam distillation is the most common method to extract orange
essential oils (Pauline, 2015). Commonly, orange EOs steam distillilation is operated at 100°C
yielding 4,4% of essential oil (Frassinetti, 2011).
Nanoparticle and Nanochitosan
Nanoparticle is the term given to particle sized between 1 and 100 nanometres (nm) with a
surrounding interfacial layer. Nanoparticles display unique physical and chemical features because
of effects such as the quantum size effect, mini size effect, surface effect, and macroquantum
tunnel effect (ramezani2015). Nanoparticles of chemical substance has a great scientific interest
for its increased surface area compared to those of normal-sized particles (Gad 2016). A large
group of studies includes the implementation of nanomaterials and nanotechnologies to create new
antibacterial agents that increase effectiveness and efficiency (Vijayalakshmi 2016).
Chitosan has been explored as a material choice to produce nanoparticles in the last years due to
its biodegradability and biocompatibility. Nano-sized chitosan or nanochitosan has been proven to
be a stronger antimicrobial agent than chitosan. Aliasghari et al. whom compared the antibacterial
efficiency of chitosan and nanochitosan on cariogenic streptococci provide a proof to support
previous statement. Different methods can be applied to obtain nano-sized chitosan but the
selection of the methodology used depends on the desired nanochitosan characteristics.