METHODOLOGY
Materials
Procurement of Materials
The Pineapple biomass which is the fibrous product of the pineapple fruit
will be collected from the by-products of the production of Pineapple-Tomato
Powered Juices of Food Research Development Center. The reagents such as
food grade chitosan powder, beeswax, shellac, sodium alginate and gellan gum
which will be used in this research will be purchased from Cagayan De Oro City
and Davao City. Other materials, equipment and apparatuses weighing scale,
thermometer, pH will be provided by the researcher.
General Experimental Approach
The general experimental approach for the study, biodegradable and
water-resistant packaging material is shown below in Figure 2. More detailed
procedure is stated on the following pages.
Making Paper from fibrous pulp
The method in making paper from fibrous pulp will be adopted from
Lewkittayakorn, et.al., 2020. According to their research, each pulp will be
mixed with a 50 g/L starch solution and blended in a blender for 5 min. The
blended ingredients will be placed on the mesh frame of 40 × 40 cm size to
make a paper sheet. The paper sheet will then be dried for 6 h in a hot air oven
at 65°C. The paper sheet will be cut to 20 × 20 cm to test packaging forming
using a compression molding machine with temperature set at 200°C and
pressure at 200 kg/cm2 for 10 min; in which this step is referred to as
“hotpressing”.
Preparation of bio-coating solutions
Five bio-coating solutions will be used to coat the fibrous paper, namely
beeswax, chitosan, shellac, alginate/gellan gum solution, and beeswax–
chitosan emulsion.

Beeswax: 20 g beeswax will be heated in a 70◦C water bath to
completely molten state.

Chitosan solution: 3 g of chitosan will be dissolved in 100 mLof
dilute acetic acid (1%v/v) and stirred until complete dissolution.

Shellac solution: this will be in completely dissolved state.

Sodium alginate/gellan gum solution (NaAlg/gellan): This method
will be adopted from Zhang, Xu, Gao, Fu, & Zheng, (2017).
2%(w/v) sodium alginate (NaAlg) and 1% (w/v) gellan powder will
be dissolved with 0.5% (w/v) glycerol in distilled water. Then the
solution will then be heated and stirred at 70–80◦C. After that, the
solution will be degassed by sonication.

Beeswax–chitosan emulsion: 1 g chitosan will be dissolved in 100
mL of dilute 1% acetic acid solution before adding glycerol to the
solution at 4:1 (v/v) ratio. Then beeswax will be added in the
chitosan solution which was heated to around80◦C in a water
bath for melting the beeswax. The weight of beeswax added must
be precise such that it will contribute 30 wt% of the emulsion film
dry matter. After that, the hot beeswax–chitosan latex will be
cooled to room temperature. This method is according to the
study of Zang, Xiao, and Qian, 2014.
Paper Surface Coating
The paper samples will be coated using a wire rod coater at constant
forward speed to ensure wet film thickness of 250 𝜇m with all coating solutions.
The samples will then be dried in an oven (65°C) for 24 h. However, in the case
of sodium alginate/gellan solution, before drying the coated paper in an oven,
it will be sprayed with 5% (w/v) calcium chloride (CaCl2) solution.
Physical and Mechanical Properties Testing
The physical and mechanical properties testing of the product will be
adapted from Ewkittayakorn, et.al., 2020. Listed below are some of the tests
that will be utilized in the assessment of the product.
Thickness Grammage
The thickness of the paper will be determined using a Vernier
caliper. Eight locations on each sample will be measured, and the mean
thickness will be determined with an accuracy of ± 0.001 mm. Whenever
possible, each test piece shall have an area of not less than 500 cm2 and not
more than 1000 cm2. The grammage in grams per square meter will be
calculated as follows:
Eq. 1
g = (m/A) x 1000
where m is the mass of the test piece (g) and A is the area of the test piece
(cm2).
Absorbency
The absorbency will be measured by placing a paper sample on
top of a cavity. 0.01 cm3(10 𝜇L) of water was dropped on top of the paper using
a micropipette. A stopwatch wasimmediately started when the preset volume
of water was dropped on the paper, and it was stopped when the water droplet
was completely imbibed into the paper. This test will be repeated five times at
different locations on each sample.
Water absorptiveness
The Cobb method will be used to determine water absorptiveness of the paper. Five samples of each type of bio-coated paper will be kept
in the conditioning atmosphere throughout the test. After weighing, the samples
will slowly be immersed in 100 ± 5 ml water (or proportionately less for a smaller
test area) in a cylinder providing 10 mm head space, and the timer was
immediately started. Fresh water was used for each determination. The
samples will be taken out of the liquid, blotted with an absorbent paper to
remove excess water, and weighed again. The procedure will be repeated until
reaching equal masses in two consecutive measurements (during 4 days), and
the absorption capacity of the paper will be calculated using Eq. (2):
Eq. 2
water absorptiveness = (𝑚2 − 𝑚1 )𝐹
where m2 is the wet mass of the test piece (g), m1 is the dry mass of the test
piece (g) and F is 10,000/test area (fora typical test apparatus this is 100 cm 2).
Burst Strength
Bio-coated paper samples (2.5 × 2.5 in.) will be used to test burst
strength. The specimens were securely clamped in position, with overlap at all
points. Hydrostatic pressure was increased as specified until the specimen
ruptured, and the maximum pressure was registered. If any movement of the
unclamped margin of the specimen was observed, that test run was rejected
and clamping pressure was increased. If, however, excessive clamping
pressure damaged the specimen, the test result was also disqualified and the
clamping pressure was reduced. Each side of the paper was tested ten times.
Internal Tear Resistance
The tear resistance of paper will be determined from the average
tearing force in accordance with the Elmendorf method. The test pieces will first
be conditioned in a controlled atmosphere. The two sides of the paper will be
distinctly marked for distinguishing between them. The bio-coated paper will be
cut to four rectangular sheets of the same size, between 50 ± 2 mm and 76 ±
2 mm wide and 43 ± 0.5 mm long. A pendulum (with augmenting mass) will be
used to regulate the energy input and to show the remaining energy by swing
amplitude. The mean readings were arranged to within the range 20% to 80%
of the full-scale reading by adjusting the mass and the number of sheets tested
in a single run. The tear resistance will be determined as follows:
𝐸𝑞. 3
𝐹=
⇀ 𝐹𝑝
𝑛
F is the tearing resistance (mN), ⇀F is the mean scale read-ing (mN), p is the
number of sheets torn simultaneouslyfor which the pendulum scale has been
calibrated to give adirect tearing resistance reading (mN), and n is the
numberof sheets torn simultaneously.
Determination of Biodegradation
Utilizing the method of determination of the biodegradation of the fibrous
paper packaging material by Ong, & Sudesh, 2016 on their study on the effects
of polyhydroxyalkanoate degradation on soil microbial community. The method
will be using analyzing each sample and it will be retrieved every 15 days from
where they were buried under soil. Each piece will be cleaned from soil debris
attached to it. The dry samples will be placed in a desiccator until a constant
weight will be reached. The weight of each sample will be recorded. The
degradation of a sample is expressed as percentage of weight loss which can
be calculated using the formula below in Eq 4:
Eq 4.
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑠𝑎𝑚𝑝𝑙𝑒−𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒 𝑎𝑓𝑡𝑒𝑟 𝑑𝑒𝑔𝑟𝑎𝑑𝑎𝑡𝑖𝑜𝑛
% weight loss = (
𝑤𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑖𝑛𝑖𝑡𝑖𝑎𝑙 𝑠𝑎𝑚𝑝𝑙𝑒
) 𝑥100
Statistical Analysis Design
Two-way analysis of variance (ANOVA) will be utilized to analyzed the
instrumental and sensory parameters to evaluate significant differences among
the samples at 95% confidence level according to Tukey’s test.