Biopolymer films from marine resources

A. Production of chitosan films

B. Chitosan film testing
Laboratory Life Sciences Processing Biological Resources Version 1.0_2022
Instructors Prof. Dr. Petra Blitgen-Heinecke Dipl.-Ing. Carlos Perez Bolde
Author of the Script Dipl.-Ing. Carlos Perez-Bolde
Last Name, First Name Student ID Signature on Experiment Day
(Please write legibly) A B

The practical trainings “Biopolymer films from marine resources” consists of two experiments, each
of which is to be conducted in a different session and in the marked order. The first part concerns
the production of chitosan films and the second part the mechanical testing of the films.

Shall you have any doubts, please do not hesitate to contact the instructors.

Please be advised that BEFORE you are allowed to conduct any activity in the laboratory, ALL the
following prerequisites MUST be covered:

• Taking the general safety briefing and being in possess of a lab coat
• Reading and understanding the information in this printout
• Submitting the answers to the preliminary questions latest one day before the practical

Additional remarks:

- Be on time, otherwise you can be denied entrance (!)

- Follow the indications of the instructors.
- Please work as tidy as you can, as you will have to clean up afterwards.
- Improvement is a continuous process, so feedback and constructive critique are most welcome.

Every group should prepare a report on the entire practical training!

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 Index of Contents
Introduction............................................................................................................................................. 3
Chitin ....................................................................................................................................................... 3
Chemical structure .............................................................................................................................. 3
Sources ................................................................................................................................................ 3
Possible uses of the raw material........................................................................................................ 4
Production ........................................................................................................................................... 5
Chitosan................................................................................................................................................... 6
Chemical structure and properties...................................................................................................... 6
Production ........................................................................................................................................... 6
Uses ..................................................................................................................................................... 7
Chitosan films .......................................................................................................................................... 7
Overall aim of the laboratory practical training ...................................................................................... 8
Part A. Production of chitosan films........................................................................................................ 9
3.1. Lessons to be learned .............................................................................................................. 9
3.2. Objectives ................................................................................................................................ 9
3.3. Chemicals................................................................................................................................. 9
3.4. Material ................................................................................................................................. 10
3.5. Procedure .............................................................................................................................. 10
Operating Directive ........................................................................................................................... 12
Part B. Chitosan film testing & Own application ................................................................................... 14
1. Lessons to be learned .............................................................Fehler! Textmarke nicht definiert.
2. Objectives ...............................................................................Fehler! Textmarke nicht definiert.
3. Procedure ...............................................................................Fehler! Textmarke nicht definiert.
4. Own application .....................................................................Fehler! Textmarke nicht definiert.
Operating Directive ............................................................................................................................... 17
Annex 1. Preparation Questions ........................................................................................................... 18
Annex 2. Report Guidelines ................................................................................................................... 18
Annex 3. Sources ................................................................................................................................... 20

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 Introduction

Chitin

Chitin is a linear polymer and the second most abundant natural biopolymer on Earth after cellulose.
It is elastic and flexible, and its structure is highly similar to that of cellulose. Chitin occurs both in plants
and animals, for instance, in fungi and in arthropods e.g. insects and crustaceans. In nature, chitin is
particularly relevant in terms of structural and defensive properties as it shapes and stabilizes
exoskeletons, protects soft parts against mechanical forces, and shelters body liquids. In fact, the name
“Chitin” comes from the Greek word chiton meaning “a coat of nail.” The industrial uses of chitin
include cosmetics, textiles, water treatment and biomedicine.

Chemical structure

Chitin or β-(1→4)-linked N-acetyl-D-glucosamine is a long linear polysaccharide and a high-molecular-

weight biopolymer of nitrogen-containing monomers. The chemical structure and, thus, the properties
of chitin and cellulose are similar. Both are derivatives of glucose and differ only in the acetamido
(–NH-C(O)CH3, in chitin) and hydroxyl (–OH , in cellulose) groups in position C2. Figures 1a and 1b
respectively show a 2-monomer 1 section of chitin and cellulose.

a) Chitin b) Cellulose

Figure 1. A 2-monomer section of (a) Chitin and (b) Cellulose.

The n after the square brackets indicates that the polymer has an n-sections chain-length;
numbers 1 and 4 stand for the position of the bond between the two monomers.

Sources

In fungi, chitin appears in the cell walls as the principal major fibrillar polymer. Chitin amounts for up
to 45% of lower fungi like oomycetes, yeasts and molds (Fig. 2a). There is also a reasonable amount of
chitin in certain higher fungi such as Basidiomycota (Fig. 2b) and Boletus edulis (also known as penny
bun or porcino).

1
Monomers are the structural basic unit of the respective polymer.

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 a) Spinellus fusiger growing on a b) Amanita pantherina,
Mycena haematopus mushroom from the Basidiomycota

Figure 2. Fungi with a high amount of chitin.

Photos: Wikipedia1, Wikipedia2

Large amounts of chitin are also present in the exoskeleton of crustaceans and in insects. Animals of
the Class Crustacea, whose exoskeleton may consist of up to 40% chitin, include crabs, lobsters,
crayfish, krill, woodlice, barnacles, and shrimps (Fig. 3a). The wings of many insects are also partly or
fully built up of chitin, such as the hindwings of the maybuck in Figure 3b.

a) Abludomelita obtusata (A crustacean) b) Maybuck or cockchafer (An insect)

Figure 3. Arthropods (animals) with a high amount of chitin.

Photos: Wikipedia3, Wikipedia4

Possible uses of the raw material

Dried crustacean shells are sold for animal-feed supplement, bait and fertilizer. Yet, the crustacean
shells may yield a larger potential due the harbored chemicals: protein (20–40%), calcium carbonate
(20–50%), and chitin (15–40%).

Protein from shells is good for animal feeds as it may contain, as in the case of Penaeus shrimp shells,
all the essential amino acids and a nutritional value comparable to that of soya-bean meal.

Calcium carbonate is widely used in the pharmaceutical, agricultural, construction and paper
industries. The current geological sources contain, however, heavy metals that may be toxic and are
difficult to remove. A heavy-metal-free source of calcium carbonate is shrimp chalk. The market price
(2015) of ground calcium carbonate was about 60 USD/ton for coarse particles (used in construction,
pigments, fillers and soil treatment) and up to 14,000 USD/ton for ultrafine particles (used to improve
the properties of rubber and plastics). 2

2
Nature 524, 155–157 (13 August 2015); doi:10.1038/524155a

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Chitin. Nitrogen-containing compounds are widely used for the fixation of carbon dioxide (CO2) and in
the pharmaceutical, cosmetic and textile industries. Ethanolamine (ETA) is used in power plants for
carbon dioxide sequestration and in skin-friendly soaps, household cleansers and surfactants. With
carbon, nitrogen and oxygen already bound in chitin, and requiring only one step to produce ETA;
chitin might be more suitable to producing ETA than the current energy intensive process.

Production

The industrial production of chitin is done by processing

crustacean shells, a by-product of the food industry. Every
year, 6 to 8 million tons of waste crab, shrimp and lobster
shells (Fig. 4) are produced globally. In fact, only 40% of a
crab's mass is meat; whereas in tuna fishes meat accounts
for 75% of the weight. The richest potential source of
crustacean chitin is krill. 200 – 800 million tons of the small
shrimp-like animals are distributed worldwide. A total catch Figure 4. Lobster shells.
of 1.5 million tons per year is allowed. Source: (Yan & Chen, 2015)

The unit operations 3 involved in the processing of chitin are listed below. Pretreatment of the raw
material, here grinding.

1. Separating the different components out, i.e. fractionation.

2.1. Demineralization: decomposition of calcium carbonate (CaCO3) with hydrochloric acid
(HCl).
2.2. Washing until neutralization, that is, until a pH value is 7 achieved.
2.3. Deproteination: removal of proteins with sodium hydroxide (NaOH) solution at elevated
temperature.
2.4. Washing until neutralization.
2.5. De-coloration (optional): color removal with de-coloring agents.
2. Drying in an oven.

In a nutshell

HCl  Removes calcium

NaOH  Removes proteins
H2O – Washing

Due to the required corrosive and hazardous solvents and the intensive use of water, high quality chitin
cost around 200 USD/kg.

Anyhow, the widespread use of chitin is prevented by two reasons. First, chitin has a very low solubility
in water, bases, acids and most organic solvents. Second, the handful of substances that do dissolve
chitin are toxic. Thus, applications of chitin are very limited. A much more interesting substance that
derives from chitin is chitosan.

3
A unit operation is a basic step in a process e.g. grinding, homogenization, pasteurization, and packaging

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 Chitosan

Chitosan is the simplest and least expensive derivative of chitin. It occurs in some fungi, but its content
is much less than that of chitin. Chitosan is industrially obtained by treating chitin or its wastes. The
quality and properties of chitosan such as purity, viscosity, molecular weight and de-acetylation degree
depend on factors such as concentration of the alkali used in the production process, treatment time,
chitin-to-alkali ratio, temperature, type of chitin source and particle size. Chitosan is commercially
available as flakes, powder and processed products like beads and films. It has antibacterial, antifungal
and antiviral properties.

Chemical structure and properties

Chitosan is a linear polysaccharide of randomly-distributed β-(1→4)-linked N-acetyl-D-glucosamine

and D-glucosaminede-acetylated units. The acetamido (–NH-C(O)CH3) group in chitin (Fig. 1a) is
replaced by a primer amine group (–NH2) in chitosan. Figure 5 shows a 2-monomer section of chitosan
with such replacement marked in red.

Figure 5. A 2-monomer section of chitosan.

The n after the square brackets indicates that the polymer has an n-sections chain-length; numbers 1
and 4 stand for the position of the bond between the two monomers.

In fact, the main difference between chitin and chitosan from a process engineering point of view is
their degree of de-acetylation. That, in turn, strongly affects their solubility, as well as the extent of
swelling in water and of biodegradation. There exist several types of chitosans with different degrees
of deacetylation as well as different degrees of polymerization.

Chitosan has a high solubility (at pH values < 6) in aqueous solutions of organic and some inorganic
acids. It has high water-retention capacity, good miscibility, high adhesiveness, chelating and sorption
properties, controlled biodegradability, and is non-toxic.

Production

The main production process of chitosan is the de-acetylation of chitin with a high concentrated alkali,
e.g. sodium hydroxide (NaOH, 45%) at elevated temperatures (~ 100°C). In this process, chitin and the
concentrated alkali react to chitosan and acetate. The reaction proceeds rapidly at the beginning, i.e.
after one hour the product is 68% de-acetylated, and afterwards it slows down, i.e. after 5 hours the
product is 78% de-acetylated.

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 In a nutshell
100°C
NaOH conc. �⎯⎯⎯� De-acetylation
After 1 hr.: 68 % de-acetylated
After 5 hr.: 78 % de-acetylated

Another way to obtain chitosan is by reacting chitin with a concentrated acid to produce chitosan and
acetic acid.

If the de-acetylation process is to be used, the unit operations involved are listed below. Please note
that the order in which they are to be performed is not random!

1. De-acetylation with concentrated sodium hydroxide (NaOH) solution at elevated temperature.

2. Washing with as little water as possible until neutralization.
3. Drying in an oven.

Uses

Chitosan has a wide range of applications. Due its sorptive properties, chitosan is used as filtration
material, flocculant and coagulant agent to remove metal ions in water treatment. In medical
applications, chitosan is found in bandages, sponges, membranes, contact lenses, artificial skin and in
dental inhibition agents because of its antibacterial, antifungal and antiviral properties. Chitosan is also
used in cosmetic products such as make-up powder, nail polish, moisturizers, creams and toothpastes.

Other applications of chitosan include enzyme immobilization, protein separation and cell-recovery in
biotechnology. Chitosan is also used for seed coatings, and for fertilizer and agrochemical release in
agriculture; and as preservative and color stabilizer in the food industry. Chitosan is also embedded in
textiles such as surgical threads, fibers and membranes.

Chitosan films

Chitosan films are biological degradable, biocompatible, non-toxic, permeable-to-oxygen, and have
high strength, flexibility and water absorbance.

Plasticizing agents may be added in small amounts to facilitate the film forming. Such agents are low
molecular weight compounds (e.g. glycerol, urea) that act upon the hydrogen bonds between polymer
chains and modify the degree of chain ordering. Pure films with high strength result from the tight
packing of the chains during the film building.

The properties of chitosan films may also be tailored by changing the crosslinking of chitosan in films.
For instance, adding bioactive components enhance the medical properties and make films degradable
by human enzymes, but the mechanical properties would in turn be lowered. Chitosan films with
bioactive components can be applied directly on the skin for treating wounds and burns.

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 Overall aim of the laboratory practical training

Starting with chitosan powder, extracted from biological sources and a monomer for this practical
training’s purposes, 5 different biopolymer films are to be produced varying in 2 factors:

• The biopolymer-to-solvent ratio, and

• The presence or absence of plasticizing agent.

After producing the 5 different biopolymer films, their properties are to be contrasted.

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 Biopolymer films from marine resources

Part A. Production of chitosan films

3.1. Lessons to be learned

• Handling of diluted acids

• Using a plasticizing agent
• Applying simple laboratory techniques
• Contrasting the quality of different products
• Identifying cause-effect relationships

3.2. Objectives

Producing chitosan films with and without plasticizing agents and contrasting their properties.

3.3. Chemicals
In a nutshell
a) Chitosan [(C6H11NO4)n, powder]
b) Acetic acid (CH3COOH, 2M) Acetic acid: Solvent
c) Glycerin (C3H8O3, technical grade) Glycerin: Plasticizing agent

Safety is a top priority when working with chemicals! The chemical hazards in this laboratory training
are: corrosive to metals (H290), skin irritation (H315) and serious eye irritation (H319). The codes of
the precautionary measurements are listed in Table 1 and explained in the Operating Directive T-001-
E-PBRc (pages 13 and 14). Please become familiar with such measurements!

Table 1. GHS of the chemicals in the laboratory training “Production of chitosan films”
Globally Harmonized System of Classification,
Substance
Labelling and Packaging of Chemicals (GHS)
CAS Hazard
Name Precautionary statements Pictograms
Number statements
Chitosan 9012-76-4 None None None
H290, H315, P280, P302+P352,
Acetic acid (2M) 64-19-7
H319 P305+P351+P338
Glycerin 56-81-5 None None None

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3.4. Material

Glassware Apparatus
• 2x Beaker, 50 mL • 1x Balance (Scales)
• 1x Beaker, 150 or 250 mL • 1x Heating plate, with
• 1x Graduated pipette, 10 mL - Magnetic stirring bar, and
• 3x Petry dishes - Temperature sensor
• 1x Stirring rod
• 1x Water bath

Miscellaneous

• 1x Flip style pipette filler (Peleusball)

• 1x Pair of tweezers • 1x Spatula

3.5. Procedure

Temperature
1. Set up your apparatus according to figure 6!

sensor
2. Start heating the water bath by adjusting the
temperature sensor to 45°C. a)
3. Pour 10 mL of acetic acid (2M) in a 50 mL beaker.
4. Add a magnetic stirrer and heat to 35°C.
5. Add chitosan powder and, if provided, glycerin and/or
vegetable oil according to the recipes given in table 2!
Stir vigorously!

b)

Figure 6. Apparatus for producing the

chitosan films. a) Steps 1 and 2, and b)
up to Step 5.

Table 2. Educts (i.e. reactants and ingredients) of the five chitosan films to be produced
Experiment Chitosan Vinegar (Acetic Acid) Glycerin Vegetable oil
1 0.2 g 10 ml - -
2 0.4 g 10 ml - -
3 0.2 g 10 ml 2 drops -
4 0.2 g 10 ml - 1 drop
5 0.2 g 10 ml 2 drops 1 drop

Note: The powder shall not stick neither on the beaker nor on the temperature sensor.

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 6. As soon as the powder dissolves, lower the stirring to a gentle rate.
7. Stir the mixture until a viscous emulsion is formed (Fig. 7).
8. Pour the viscous solution onto the lid of a Petri dish.
9. Fill in the respective part of Table 3 "Properties of the wet films".

10. Proceed with the next experiment according to table 2.

11. After having prepared all five films, make photos of the wet films and
allow the solvent, i.e. the acetic acid, from all five wet films to Figure 7. Desired
vaporize undisturbed for 2 to 3 days so that the film can dry and consistency of the emulsion
harden.

Table 3. Properties of the wet chitosan films

Films
Chemicals & Ingredients 1 2 3 4 5
Chitosan 0.20 g 0.40 g 0.20 g 0.20 g 0.20 g
Vinegar (Acetic acid) 10 ml 10 ml 10 ml 10 ml 10 ml
Glycerin (Plasticizer) - - 2 drops - 2 drops
Oil - - - 1 drop 1 drop
Properties 1 2 3 4 5
Thickness

Solids content

Bubble presence

Resistance to flow

Key:
Thickness: Very thin – Thin –thick – Very thick Bubble presence: None – A few – Some – A lot
Solids content: None – A few – Some – A lot Resistance to flow: Low – Middle – High

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 Number: T-001-E-PBRc
Operating Directive
Page 1 of 2
acc. to GefStoffV
Faculty Life Sciences Working place/ Field of activity: Laboratory 10 02 013
Procedure of the Experiment

Processing Biological Resources:

Production of chitosan films

Hazards for humans and the environment

• Acetic acid (2M) is a diluted acid that:
a) H290 - May be corrosive to metals.
b) H315 - Causes skin irritation.
c) H319 - Causes serious eye irritation.
• Risk of burning:
a) If mishandled, the magnetic stirrer and the water bath can be hot. The
experiment is to be performed at 35°C.
• Risk of being cut:
a) If mishandled, glassware can shatter, and result in flesh wounds.
Safety measures and codes of conduct
• Reading the present document and passing a pre-experiment test are
required to conduct the experiment.
• Perform the experiment according to the instructions.
• Proceed in a proper, orderly way.
• Acetic acid (2M):
a) P280: Wear protective gloves, protective clothing, and eye protection.
Protective gloves EN 374 tested: Material: NBR (Nitrilkautschuk),
>0,11mm thick, permeation level 6.
b) Work under a chemical fume hood.
c) Avoid skin and eye contact with the chemicals. Do not inhale the
vapors.
d) Small, closed bottles with 90 mL of acetic acid (2M) are to be provided
(80 mL are required for the experiment).
Conduct in emergencies Emergency call: 112
• Spillage of Acetic acid (2M)::
a) Spillage of small amounts: Clean with a sufficient amount of water.
b) Spillage of large amounts: Absorb the spillage with absorbent
(Chemsorb), and dispose as solid waste. Evacuate area. Warn affected
surroundings.

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 Number: T-001-E-PBRc
Operating Directive
Page 2 of 2
acc. to GefStoffV
Faculty Life Sciences Working place/ Field of activity: Laboratory 10 02 013
Procedure of the Experiment

Processing Biological Resources:

Production of chitosan films

First aid Emergency call: 112

• Take injured persons out of danger.
• Provide first aid, paying attention to self-protection.
• Call first aid (-299), call ambulance.
• Instruct rescue personnel.
• Acetic acid (2M):
a) Skin, hair, clothing contact (P302+P352): Take off immediately all
contaminated clothing. Rinse skin with plenty of cold water.
b) Eye contact (P305+P351+P338): Rinse open eye cautiously with
water for at least 15 minutes. Remove contact lenses, if present and
easy to do.
• Burns:
a) Immediately remove clothing affected by hot substances.
b) Submerge affected body parts in cold water or place under running
cold water immediately, until pain relief sets in.
c) Leave burns affecting face and eyes undressed.
• Cuts:
a) Use disposable gloves and then a first-aid kit.
b) Stop bleeding where possible.
c) Apply dressings.
• Do not leave injured people unattended until the rescue service arrives.
• In the event of an accident and subsequent visit to a medical doctor or
facility, there is the obligation to inform the Hochschule as soon possible.
Proper disposal
• Carefully neutralize under cooling the excess acetic acid with sodium
hydroxide.
• Liquid waste must be disposed into the inorganic waste container.
• Solid waste must be disposed into the solid waste container.

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 Part B. Chitosan film testing

Before you start the testing of the mechanical properties of the films, first of all, describe their
appearance and enter the attributes in the following table! Make photos of the dry films!

Table 4. Properties of the dry chitosan films

Films
Chemicals & Ingredients 1 2 3 4 5
Chitosan 0.20 g 0.40 g 0.20 g 0.20 g 0.20 g
Vinegar (Acetic acid) 10 ml 10 ml 10 ml 10 ml 10 ml
Glycerin (Plasticizer) - - 2 drops - 2 drops
Oil - - - 1 drop 1 drop
Properties 1 2 3 4 5
Color

Surface

Defects

Key:
Color: Transparent – Almost transparent – Yellow Surface: Straight – Slightly bent – Bent
Defects (choose all relevant): Cracks, Orange peel, Trapped bubbles

Preparing the testing

1. Remove the dry film from the Petri dish’s lid. Test 1dry
Test 2dry
2. With a pair of scissors cut the dry film in 4 parts, as the dotted
lines in Figure 12 show. Test 1 moisty
Test 2 moisty

Figure 9. Parts of a dry film for

Tests the selected tests.

Tearing means breaking apart by force without using cutting tools. Even resistant-to-tearing
materials may be easy to tear when compromised, for instance by a small crack, cut or tear.

Elasticity is the ability to resist distortion under load and return to the original shape and size. If a
solid material returns to its initial shape and size upon removal of the load, it is elastic. On the
other hand, if the material fails to do so and remains deformed, it is plastic.
Please note that the content of acetic acid in the produced films is negligible, as it evaporated.
The only components in your film are, thus, chitosan, vegetable oil, glycerin and food colorant
(optional), all of which are completely safe. So, no gloves are needed for handling the films.

For each film that you are testing proceed as follows. The order is important.

Dry films

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 Test 1a. Tearing resistance (dry film)

3. Take Part 1a of the film (Fig. 9) and try tearing it apart using your fingers or a tweezer.

Assign the tearing resistance on a scale between 0 and 10 (where 0 means worst and 10 best)

Test 2a. Elasticity (dry film)

4. Proceed like in 3.

Assign the elasticity on a scale between 0 and 10 (where 0 means worst and 10 best)

Moist films

5. Put water to boil.

6. When the water starts boiling, hold a piece of film (Fig. 9) with the wooden clamp over the
water vapor for 5 seconds. It should get moist.

7. Let the film cool down to room temperature trying to avoid the evaporation of the tiny water
bubbles that were formed on the piece of film.

Test 2a. Tearing resistance (moist film)

8. With the moist film, proceed like in 3.

Consider: What changed in terms of tearing resistance compared to the dry film (Test 1a)?

Test 2b. Elasticity (moist film)

9. Proceed like in 3.

Consider: What changed in terms of elasticity compared to the dry film (Test 2a)?

Test 3. Waterproofness

10. At the end, drip a few drops of water directly onto the remains of one of the dry films tested

Consider: What happens to the film? Are there holes?

11. Repeat steps 1-3 with the other films.

Table 5. Chitosan film testing

Films
Chemicals & Ingredients 1 2 3 4 5
Chitosan 0.20 g 0.40 g 0.20 g 0.20 g 0.20 g
Vinegar (Acetic acid, solvent) 10 ml 10 ml 10 ml 10 ml 10 ml
Glycerin (Plasticizer) - - 2 drops - 2 drops
Oil - - - 1 drop 1 drop
Dry film 1 2 3 4 5

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Tearing resistance (1-10)

Elasticity (1-10)

Moist Film 1 2 3 4 5
Tearing resistance (1-10)

Elasticity (1-10)

Waterproofness 1 2 3 4 5
Waterproofness (yes/no)

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 Number: T-001-E-BR_LtG
Operating Directive
Page 1 of 1
acc. to GefStoffV
Faculty Life Sciences Working place/ Field of activity: Lab 10 02 013
Procedure of the Experiments

Processing Biological Resources:

Chitosan film testing
Hazards for humans and the environment
• Risk of burning:
a) The kettle (or pot) and the apparatus are hot during the process.
b) There is hot water (60°C) in the water bath.
c) During the process water vapor is produced
• Risk of being cut:
a) If mishandled, glassware can shatter, and result in flesh wounds.
Safety measures and code of conduct

• Experiments are not suitable for children & shall not be conducted in their presence
• Reading the present document and passing a test are required to do the
experiment
• Perform the experiment according to the instructions.
• Proceed in a proper, orderly way.
• Kettle, pot, and water bath shall neither be touched nor be transported while hot.
• Use suitable protective gloves to handle hot glassware.
• Use the wooden clamp to fix glassware and to expose the films to water vapor.

Conduct in emergencies & First aid Emergency call: 112

• Take injured persons out of danger.

• Provide first aid, paying attention to self-protection.
• Call first aid (-299), call ambulance, and instruct rescue personnel.
• Burns:
a) Immediately remove clothing affected by hot substances.
b) Submerge affected body parts in cold water or place under running cold
water immediately, until pain relief sets in.
c) Leave burns affecting face and eyes undressed.
• Cuts:
a) Use disposable gloves and then a first-aid kit.
b) Stop the bleeding where possible.
c) Apply dressings.
• Do not leave injured people unattended until the rescue service arrives.
• In the event of an accident and subsequent visit to a medical doctor or facility,
there is the obligation to inform the Hochschule as soon possible.

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 Annex 1.

Preparation Questions

Questions to be ansered

1. What is the second most abundant natural biopolymer on Earth after cellulose?
2. The name comes from the Greek word chiton meaning “a coat of nail.”
3. Name two applications of chitosan!
4. Which properties of chitosan are important for water treatment applications?
5. In which products can chitosan be found? Give examples.
6. What is the main difference between chitin and chitosan?
7. What is the main raw material source for producing chitin and chitosan?
8. What are Plasticizing agents?
9. What are Plasticizing agents used for?
10. Why are bioactive components added to chitosan films?
11. In the production of chitosan films (Experiment C), ACETIC ACID (CH3COOH, 2M) is used as
.

12. In the production of chitosan films (Experiment C), GLYCERIN is used as

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 Annex 2. Report Guidelines

A report per group must be submitted. The report must include:

1. The signed cover page of this document.

2. The answers to the Preparation Questions in Annex 1.

3. The completed following tables:

• Table 3. "Properties of the wet chitosan films"
• Table 4. "Properties of the dry chitosan films"
• Table 5. "Chitosan film testing"

4. The following photos:

• Wet films (all 5 in a single photo)
• Dry films (all 5 in a single photo)

12.Based on the information in the previous two points, identify cause-effect relationships between:
• The properties of the wet films depending on the recipe, and
• The properties of the dry films depending on the recipe (max. 1 page)

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 Annex 3. Sources

Journal
Yan, N. and Chen, X. (2015). Sustainability: Don't waste seafood waste. Nature 524 (7564) p. 155-157.

Photos (online retrieval)

Wikipedia1: Spinellus fusiger (Photo). Available on: https://en.wikipedia.org/wiki/Spinellus_fusiger.

Retrieved on December 09th, 2020.

Wikipedia2: Pantherpilz (Photo). Available on: https://de.wikipedia.org/wiki/Pantherpilz. Retrieved

on December 09th, 2020.

Wikipedia3: Abludomelita obtusata (Photo). Available on:

https://en.wikipedia.org/wiki/Abludomelita_obtusata. Retrieved on December 09th,
2020.

Wikipedia4: Cockchafer (Photo). Available on: https://en.wikipedia.org/wiki/Cockchafer. Retrieved

on December 09th, 2020.

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