Research seminar

Extraction, characterization and chemical

functionalization of phosphorylated cellulose
derivatives from Giant Reed Plant

Rahul Ranjan
Research Scholar
School of Biochemical Engineering,
IIT (BHU), Varanasi

1
Table of content

 Introduction to cellulose

 Extraction of cellulose microfibers

 Phosphorylation of cellulose microfibers

 Preparation of phosphorylated cellulose nanocrystals

 Conductometric titration

 Characterization of prepared material

 Results and discussions

2
 Cellulose
 First discovered in 1838 by Anselme payen.

 Cellulose present as microfibrils in plant cell wall.

 Cellulose is a linear polysaccharide consisting of several units of D-glucose

joined together with β-1,4 glycosidic linkage.

OH OH
OH
O HO O
O O
O O O
HO HO
OH n
OH
OH

Figure 1 :- Structure of cellulose Figure 2 :- Formation of glycosidic linkage

between two monomers of glucose
http://chemistry.tutorvista.com/organic-chemistry/glycosidic-linkage.html
Anselme Payen, “Mémoire sur la composition du tissu propre des plantes et du ligneux”, Comptes Rendus, 1838, 7, 1052-1056. 3
Advantages of cellulose
Abundantly
available

Renewable
Biodegradable Cellulose
biopolymer

Low density

Properties of cellulose
Chemical formula (C6H10O5)n.
Density 1.5 g / cm3
Melting point 260-270 °C
Appearance White

4
 Extraction of cellulose microfibers

Extraction of cellulose microfibres was carried out

from Giant Reed (plant) in three steps.

First step involve the grinding and sizing and washing

of biomass.

Second step includes the alkali pre-treatment of

biomass using NaOH (4%) at 80 oC for 2 hours.
Figure 3 :- extraction of
CMFs from Giant Reed
plant
Third step is the bleaching of pre-treated biomass
using glacial acetic acid, sodium chlorite and NaOH,
finally produces pure cellulose microfibres (CMFS).

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Phosphorylation of cellulose microfibres
Phosphorylation was carried out using Ammonium di
hydrogen phosphate (4.5 g) and urea (12 g) in 15 ml of
distilled water.

Prepared solution was added into the 10 g CMFS.

Prepared mixture was dried over night at 60 oC.

Figure 4 :- Phosphorylation
process
Cured at 150 oC.

Phosphorylated cellulose fibres were cooled and

repeatedly washed to remove non attached chemicals.

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Preparation of phosphorylated CNCs
1.0 g of CMFs was mixed to 20 ml of phosphoric acid and heated at 60 oC
overnight.

Then the mixture was homogenised for 2 hours.

Whole reaction mixture was put into ice bath to stop the reaction.

Reaction mixture was washed repeatedly by successive centrifugation at

12000 rpm for 20 minutes at 15 oC.

The resulting suspension was dialysed in distilled until pH reached to 7.

Sonicated in ice bath for 15 minutes .

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Conductometric titration
Charge content of phosphorylated cellulose was determine using conductometric
titration.
Steps involved :-
0.3 g of P-CMFs and P-CMCs was dispersed in 100 ml of solution containing 10
ml NaCl (0.01 M ).

O.1M OF HCl was added to the dispersed solution to bring the pH 2.5-3.

Then titration was carried out by adding 0.1M NaOH solution step wise and
conductivity was measured till the pH reached to 11.
Charge content (CC) was determine using following formula :-
CC (millimole kg-1) =
Where,
C NaOH corresponds to the concentration of NaOH.
W CNC is the weight of phosphorylated sample (g).
V NaOH is the volume of NaOH (L) corresponding to plateau region.

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Material characterization
 Chemical structure of cellulosic material was studied using Fourier transform
infrared spectroscopy equipped with attenuated total reflectance  (FTIR-ATR). All
the spectra were recorded in 4000 - 600 cm-1 range with a resolution of 4 cm-1
with accumulation of 16 scan.

 Crystalline structure of all the material were analysed using X-ray

diffractometer (XRD). All the samples were scanned with Cuk radiation in the
range of 5-40 oC.
 Voltage and current were fixed at 40 kV and 40 mA, respectively.
 Crystallinity index were calculated using Segal equation :-
X 100
Where,
I200 is the maximum peak at crystalline phase
Iam is the maximum from the amorphous phase

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Material characterization
 Effects of chemical treatments over the morphology of cellulosic fibres were
studied using scanning electron microscopy operated at 15 kV.
Samples were coated with thin conducting carbon using a sputtering apparatus.

 Atomic force microscopy (AFM) was used to investigate the morphology and
dimensions of the nanocrystals.

 The thermal stability and the degradation behavior of all samples were evaluated
by thermogravimetric analysis (TGA) using TA instruments.
 Samples were heated from room temperature to 700°C at a heating rate of
10°C/min.

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Results and discussion
Common method of grafting phosphate group on cellulose surface via
substitution –OH group by phosphate at C2, C3 and C6 sites results in
formation of crosslinked and uncrosslinked structure.

Figure 5 :- Indicating the possible cellulose structure after phosphorylation

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Conductometric titration

Figure 6 :- Representing conductometric titration curve for (A) P-CNCs and

(B) P-CMFs

Key findings
Total charge content in P-CNCs was 254 mmol kg-1
Total charge content in P-CMFs was 3133 mmol kg-1
Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 FTIR spectra analysis

Figure 7 :- FTIR spectra of raw GR, extracted CMFs, and MCC (a) and P-CMFs
with P-CNCs samples (b)
Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Analysis of spectra
Wavenumber Functional Wavenumber Functional
(cm-1) group (cm-1) group
896 -OH bending 834 P-OH
1023 -C-H deformation 1230 P-O-C
1105 -C-O stretching 910 P=O
1160 -C-O-C vibration
1320 -CH bending 2903 -CH stretching
1240 -C-O stretching 3335 -OH stretching
vibration
1370 -C-O stretching
1423 -C-H deformation Key findings
Peak at 834, 1230 and 910 cm-1 confirms
1628 -CH2 vibration the successful incorporation of
1508 -C-O of acetyl or phosphate groups in the moieties of
carbonyl group cellulose.
1731 -C-O of acetyl or
carbonyl group

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Analysis of XRD spectra

Figure 7 :- XRD patterns (A) raw GR fibers and extracted CMFs (B), P-CMFs,
P-CNCs and MCC
Key findings
Crystallinity of GR = 56%
Crystallinity of CMFs = 73%
Crystallinity of P-CMFs = 70%
Crystallinity of PCNCs = 83%
Crystallinity of MCC = 83%
Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Morphological feature

Key findings
Due to chemical treatments of
fabrics dimension reduces but
in the case of native GR its
surface is rough and long
fibres were present.

Figure 8 :- SEM images of Raw GR (a), as-extracted CMFs (b), and P-CMFs (c)

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 AFM analysis of P-CNCs

Figure 9 :- Aqueous suspension and AFM images of P-CNCs

Key findings
Needle like structure in a cellulose
nanocrystal is the confirming results
which signifying proper extraction of
CNCs from GR
Aspect ratio was found to be 33.

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 TGA anlysis

Figure 10 :- TGA (a) DTG (b) curves of raw GR fibers, extracted CMFs, P-
CMFs and P-CNCs

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Structural, morphological and thermal parameter of GR
fibres, CMFs, P-CMFs, PCNCs and MCC sample

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Conclusion
 Giant Reed (GR) plant has been valorized for the first time to produce
phosphorylated cellulosic materials.
 Cellulose microfibers (CMFs) were extracted through alkali and bleaching
treatments of raw GR, with an average yield and diameter of 37% and 13 µm
respectively.
 Then, two different phosphorylation processes with and without urea have been
performed for the preparation of phosphorylated micro (P-CMFs)and nano (P-
CNCs) cellulose.
 With urea, a higher surface charge content (CC) was found.
 The prepared P-CMFs showed a micro-sized shape with an average diameter of
15 µm and a CC of 3133 mmol kg-1.
 While the extracted P-CNCs exhibited a needle-like shape with an important
aspect ratio of (L/D) = 33 and very low CC of 254 mmol kg-1.
 FTIR analysis confirmed the total removal of non-cellulosic compounds from the
surface of microfibers, as well as their partial oxidation after phosphorylation.
 The cellulose I crystalline structure has been confirmed for all samples, with a
slight decrease in the crystallinity index of P-CMFs (70%) compared to that of
CMFs (73%) due to the presence of high CC.
 P-CNCs showed a similar crystallinity index of 83% to that of microcrystalline
cellulose (MCC). The TGA analysis showed high thermal behavior for P-CNCs
with early dehydration and high char content for P-CMFs
Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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 Continue…..
 From these results, we expect that the use of P-CNCs as reinforcement in nano
composite development will be very beneficial due to its important aspect ratio,
high thermal stability, high crystallinity and the ability of phosphate groups to
improve the antibacterial properties.
 CMFs may be useful in developing fire-retardant materials due to its high CC
leading to early thermal dehydration with rapid char formation.
 Regarding the diverse potential use of phosphorylated cellulose, it is motivating
to produce micro- and nano-cellulose from sustainable and inexpensive source
as GR plant with the aim of valorising unexploited Moroccan natural resources.

Ait Benhamou A, Kassab Z, Nadifiyine M, Salim M H, Sehaqui H, Moubarik A and El Achaby M 2021 Extraction, characterization and chemical
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Plan : Bamboo Aerogel
Delignification of bamboo
Phosphorylation of delignified bamboo
Synthesis of bamboo aerogel
Characterization

1. FTIR
2. XRD
3. SEM
4. EDS and elemental mapping
5. TGA
6. XPS
7. BET
8. Water uptake and swelling
9. Application

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 Tribology
Tribology

Tribo logy
s
Means to “to Means “to
rub” study”

 The term “tribology” was coined in late 1960s by British Mechanical engineer Dr. H.
Peter Jost.
 He published a report mentioned the implication of friction, wear and corrosion in
the GDP of UK economy
 Tribology is the science and engineering of interacting surface in relative motion.
 It the branch of material science and mechanical engineering which includes the
principles of friction, lubrication and wear.
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Tribology down the history
 Early civilization developed quite sophisticated tribological devices such as
wheel of potters, door hinges, wheeled carriage.
 Carving of the Saqqara tomb of Egypt 4400 years ago shows expertise of
tribologist bending down to lubricate the sled that carries a statue of Ti.
 In the day of Roman empire, tribological principles were used for devising war
machinery and methods of fortification.
 Warship discovered near Neri lake (Rome), contain bronze ball and rollers used
to support rotating platform.
 During the age of renaissance, Leonardo da Vinci a famous painter, sculpture
and engineer who discovered that force of friction between two moving object
proportional to the normal force.

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Tribology down the history

The British Library Releases 570 Pages of Leonardo da Vinci's Manuscripts Online 25
Friction
 Friction is the force that hinder or resist the relative motion of two bodies in
contact.
 It arises from complex molecular and mechanical interaction between the
contacting surface.
Types of friction
• Solid friction
• Fluid friction
• Mixed friction
Friction between two bodies are material independent and dependent upon:
• Size of contact or molecular adhesion
• Surface roughness
• Load or pressure on the surface
• Deformation

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 Molecular adhesion, roughness and deformation
•When two bodies are brought in contact, many atoms or

molecules from one object are in such close proximity to those in

another molecular or electromagnetic forces attract the object
together this is called force of adhesion.

•When one object trying to cross over another requires breaking of

these force of adhesion.

•Roughness of the surface is another factor which influences the

frictional forces.

•When a material move forcefully it intends to deform due to

frictional forces
27
Coefficient of friction
 Frictional force is proportional to the load, therefore
F (force) = µL(Load)
 Friction is commonly represented by the coefficient of friction “µ”. The
coefficient of friction is unit less number where F represent the frictional force
experienced by two contacting bodies in motion and “L” representing the
normal forces occurred during pressing of both the contacting bodies.
 The values of “µ” typically ranges between 0 to 1. higher the value of “µ”
higher will be the frictional forces or the resistance between two contacting
bodies toward motion.

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 Metal surface
 All the metal surface irrespective of their finish, contains ridges, peaks and valley.
They stick out the surface forming peaks and valley at microscopic level. These
peaks are called asperities.
 When two metals comes in contact, solid friction sometimes called static or
adhesive friction arises. The surface undergoes adhesion or cold welding.
 When the surface start to move, kinetic friction come into play. Kinetic friction arise
from plowing of asperities of one surface across the other.
Wear
Wear is the process of removal of material from one both the solid surface in solid
contact, occurring when these two bodies are in sliding or rolling motion.
Types of wear
Abrasian
Adhesion,
Erosion and surface fatigue
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 Abrasive wear
 Abrasive wear occurs when a harder material is rubbing against a softer material.
 If there are only two rubbing parts involved in the friction process the wear is called
two body wear.
 In this case the wear of the softer material is caused by the asperities on the harder
surface.
 If the wear is caused by a hard particle (grit) trapped between the rubbing surfaces
it is called three body wear. The particle may be either free or partially embedded
into one of the mating materials.
In the micro-level abrasive action results in one of
the following wear modes:

•Ploughing. The material is shifted to the sides of the

wear groove. The material is not removed from the
surface.
•Cutting. A chip forms in front of the cutting
asperity/grit. The material is removed (lost) from the
surface in the volume equal to the volume of the wear
track (groove).
•Cracking (brittle fracture). The material cracks in the
subsurface regions surrounding the wear groove. The
volume of the lost material is higher than the volume of
the wear track.

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Adhesive wear
Adhesion wear is a result of micro-junctions caused by welding between the
opposing asperities on the rubbing surfaces of the counterbodies.
The load applied to the contacting asperities is so high that they deform and
adhere to each other forming micro-joints. The motion of the rubbing counterbodies
result in rupture of the micro-joints.
The welded asperity ruptures in the non-deformed (non-cold worked) regions.
Thus some of the material is transferred by its counterbody. This effect is called
scuffing or galling.
When a considerable areas of the rubbing surfaces are joined during the friction a
Seizure resistance (compatibility) seizure of one of the bodies by the counter body
may occur.

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Fatigue wear
 Fatigue wear of a material is caused by a cycling
loading during friction. Fatigue occurs if the applied
load is higher than the fatigue strength of the
material.
Fatigue cracks start at the material surface and
spread to the subsurface regions. The cracks may
connect to each other resulting in separation and
delamination of the material pieces.
 One of the types of fatigue wear is fretting
wear caused by cycling sliding of two surfaces
across each other with a small amplitude
(oscillating).
 The friction force produces alternating compression-
tension stresses, which result in surface fatigue.
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Erosive wear
Erosive wear is caused by particle impingement (solid, liquid, or gaseous), which
removes material fragments from the surface due to momentum effect.
Cavitation in the lubrication oil can cause erosive wear of engine bearings. When oil
exits the convergent gap between the bearing and journal surfaces, cavitation voids
(bubbles) may form.
The oil pressure rapidly drops, creating conditions for the formation of voids (the
pressure is lower than the oil vapor pressure). The bubbles (voids) then collapse,
creating a shock wave that removes particles of the bearing material.

33
Delamination wear
Wear process where a material loss from the surface by force of another surface
acting on it in a sliding motion in the form of thin sheets.

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Stick and slip friction

Reasons:
• Sliding of one objects over another under steady pulling force.
• Sometimes occur at constant or near constant velocity.
• When friction force and sliding velocity does not remain same as a function of
time or distance produces oscillation this phenomenon is called stick and slip
friction.

35
During the stick phase, frictional force build up to certain level and once large
enough force has been applied to over come static frictional force slip occurs at
interface.
Usually saw teeth pattern observed in the stick-slip process.
In a case where the coefficient of kinetic friction is less than the coefficient of static
friction there will be exist a tendency for motion to be intermittent rather than
smooth.
Two contact surface will stick until the sliding forces reaches the value of static
friction.
Sliding friction
It occurs when two surface slides each other
Rolling friction
When an objects roll over another solid surface

36
37
 Pin on disc tribometer
Tribometer (pin A pin is held stationary and disc rotates.
A pin disc tribometer consist of stationary pin under applied load in contact with rotating disc.
The pin can have any shape to simulate the contact bur spherical tips are generally used due to
its simple geometry.
on disk test ring)
For the de termination of adhesive wear in metal-to-metal contacts.
A cylindrical sample is pressed against a rotating disc or ring with a defined
load.
The torque is measured continuously and thus the coefficient of friction µ.
As a further option it is possible to measure the sample heating.
The test can be run either dry or lubricated.
The test stand can be operated with different motor-gearbox variants.
Friction speeds of 1 cm/min to 4 cm/min or 0.5 m/s to 20 m/s at surface
pressures of up to 10 MPa (depending on the specimen geometry) can be
realized.

38
39
Tribological parameter

40
Polyimide adhesive tapes as a versatile and disposable substrate to produce
CO2 laser-induced carbon sensors for batch and microfluidic analysis
Fabrication of the laser-scribed sensors (LSS)
The sensors were produced with a CO2 laser cutter (Work Special Laser-wavelength
of 10.6 pm and a pulse duration of ~ 14 μs).

The software RDWorks 8.0 was used to engineer three-electrode electrochemical

cells.

Two Kapton adhesive tapes were glued together, generating a double polyimide layer.
This tape was carbonized under ambient conditions (27 ◦C).

The conditions were laser power = 8.0 % (~1.4 W), scan rate = 80.0 mm s- 1, Z-
distance = 13.0 mm, and distance between radiation lines = 0.04 mm.

After carbonizing, the conductive tracks were partially painted with silver ink. Silver ink
was also applied to the reference electrode.

The area of the electrochemical cell was delimited with Silicon glue (Tek Bond -
transparent silicone sticker), generating a three-electrode miniaturized system

41
 Laser-scribed sensors - Paper-based microfluidic devices (LSS-
µPAD) fabrication
The geometry was designed with CorelDRAW software.

ColorQube 8870 wax printer (Xerox, Connecticut) was used to deposit the wax (4-pt
line thick) onto the Whatman 4 qualitative filter paper.
The resulting material was placed on a plate heated at 120 ◦C for 120 s, producing the
hydrophobic barriers.

The microfluidic system has two reservoirs. The inlet reservoir is a PMMA plate with a
hole of 6 mm in diameter and 3 mm in height.
The outlet reservoir was engineered as a wicking 270◦ fan

This reservoir consists of a wax-printed semicircle (50 mm in diameter) with 4 pieces of

paper sheets.

The resulting device was sealed with packing tapes. After fabricating, the inlet reservoir was
constantly refilled with the supporting electrolyte throughout the experiments

42
Fig. 1. A) Representation of fabrication process and the dimensions of the electrochemical systems.
Representative image of the B) LSS and C) LSS-µPAD. WE: working electrode; AE: auxiliary electrode; RE:
reference electrode.

43
Fig. 2A, C, E, and G show the results obtained from the respective cyclic
voltammograms.
The resistivity of the carbon tracks was measured using a multimeter, and the values are
shown in Fig. 2 B, D, F, and H.
Notes: Cyclic voltammetry (CV) is a powerful and popular electrochemical technique
commonly employed to investigate the reduction and oxidation processes of molecular
species. 

44
 Explanation to Figure 2a
The anodic peak current (Ipa) increased from 0.02 to 0.06 mm, probably due to the increase in
the active electrochemical area of the sensors. Then, Ip decreased to 0.10 mm, indicating that
higher distances between the carbonization lines compromise the electrical conductivity.
 This parameter also impacted the peak-to-peak potential separation (ΔEp). ΔEp can be directly
related to the electrical conductivity of the resulting material demonstrating more conductive
materials are obtained for smaller distances between radiation lines (Fig. 2B). Considering the
lower resistivity of the carbon material, 0.04 mm was selected as the best condition to carbonize
the polyimide substrate.
Explanation to Figure 2C,D,E and F

Fig. S2C shows that 7.5 % and 8.0 % power generated sensors with consistent analytical signals
for Ru(NH3)6 3+/Ru(NH3)6 2+. < 7.5 % powers were not enough to completely carbonize the
polyimide, generating carbon paths with high resistivity (Fig. 2D). > 8.0 % power partially damaged
the polyimide substrate, increasing the resistivity and restricting the sensor fabrication.
Considering that 8.0 % power provided better electrical conductivity (Fig. 2D), this value was used
to produce the sensors. Besides this parameter, the optimization study evaluated the scan rate.
 Fig. 2E shows Ipa slightly increased from 80 to 140 mm s - 1. ΔE p was close to 75 mV regardless
of the carbonization scan rate. Even though the scan rate did not significantly impact the sensor’s
electrochemical response, the resistivity of the carbon path is
increased by increasing this parameter (Fig. 2F).

45
 Magnitudes of the anodic and
cathodic peak currents (ipa and ipc
respectively) and of the anodic and
cathodic peak potentials (Epa and
Epc respectively).
 Faradic current is a short duration
interrupted direct current with a
pulse duration ranging from 0.1 to 1
ms with a frequency of 50 to 100 Hz

46
This result demonstrates higher scan rates might not effectively carbonize the polyimide
substrate, resulting in materials with poorer electrical conductivity. Hence, 80 mm s- 1
was selected to produce the sensors.
The Z-distance, the height measured from the polyimide substrate to the laser tip, was
finally considered to fabricate the sensors.
Fig. 2G shows Ipa and ΔEp did not significantly vary with this parameter. However, the
consistency among the resistivities was remarkably enhanced by increasing the
Zdistance, achieving lower and reproducible values at 13 mm (Fig. 2H). Consequently,
13 mm was chosen as the optimum Z-distance

47
Fig. 3. Cyclic voltammograms recorded in 1 mmol/L ferrocyanide and 0.1 mol/L KCl at 50 mV s- 1.
A) Voltammetric measurements for ten different electrodes. B) Twenty voltammetric measurements
were recorded with the same electrode. C) Nyquist EIS spectra. The frequency range was from
105 to 101 s- 1 Hz. Amplitude = 10 mV. Open circuit potential (OCP): + 0.22 V. EIS measurements
recorded for (■) commercial screen-printed electrode and ( ) proposed sensors in a mixture of 1
mmol/L ferricyanide/ferrocyanide with 0.1 mol/L KCl. D) Voltammetric measurements recorded with
the (––) commercial screenprinted electrode and ( ) the proposed sensor in 1 mmol/L
ferrocyanide and 0.1 mol/L KCl at 50 mV s- 1.
Explanation to Figure 3C
 The proposed sensor was compared with screen-printed commercial electrodes. Fig. 3C shows
the Nyquist plots. The semicircle profile corresponding to the electron-transfer limited process is
observed at higher frequencies. This semicircle is followed by a straight line, representing
the diffusion-limited electrochemical process. The diameter of the semicircles is used to obtain
the charge transfer resistance (RCT) of the materials.

 Explanation to Figure 3D
The RCT values were 1.9 ± 0.3 kΩ for commercial screenprinted electrodes and 10.3 ± 2.9 kΩ
for laser-scribed electrodes. Even though LSS had higher RCT values, they provided a
voltammetric profile comparable with commercial screen-printed electrodes (Fig. 3D),
demonstrating that the proposed materials are promising candidates to be explored as
electrochemical sensors.

49
A) Image of the LSS. B) Cyclic voltammograms recorded with LSS in 0.1 mmol/L KCl and different
concentrations of [Ru(NH 3)6]3+ at 50 mV s- 1. C) Calibration curve for [Ru(NH 3)6]3+ obtained by
voltammetry. D) Image of the µLSS-PAD. E) Transient current signals for injections of 0.2, 0.4, 0.6,
0.8, 1.0 mmol L–1 [Ru(NH 3)6]3+ solution in 0.1 mol L–1 KCl. Injection volume: 2 μL. Edetection =
-0.4 V vs Silver pseudo-RE. F) Calibration curve for [Ru(NH 3)6]3+ obtained by amperometry.

50
Charge content of the film prepared after changing the
curing time

0.3 g of dried films prepared after different curing time (30 min, 60 min, 90 min and 120 min) were taken into a beaker and
dispersed in 5 mL of sodium chloride solution

Further 85 mL of of distilled water were added into the beaker to make uniform suspension of the films

Then the pH of the suspension were lowered down to 2.5-3 by adding 0.01 M HCl

Finally, the suspensions were titrated with 0.1 M NaOH solution by dropping 200 microliters after each interval of reading till
the pH reached 11

During the experiment beaker was continuously shaked to maintain the uniformity in the suspension.

Finally, the charge content were calculated using below mentioned formula
Charge content (mmol/kg)

where CNaOH is the concentration of NaOH (M) used for titration, V NaOH is the volume of
NaOH used during the titration (L) corresponds to the plateau region and is the weight of
dried film.
51
52
Tensile strength of the films
Testing condition:
Films were dried at 80°C before
the test.
Thickness = 0.02 mm
Gauge length = 25 mm
Dia = 18 mm
Area = 0.36 mm2

Sample Maximum stress Maximum strain Modulus (MPa)

(MPa) (%)
CT_30 55.76 0.145 384.55
CT_60 29 0.24 120.83
CT_90 27.99 0.134 208.88
CT_120 27.9 0.0134 1335.82
53
Biodegradability test of the films (2 hours)
Soil was taken from the agricultural farm IAS (BHU) and humidity were checked and maintained at 50% at pH 6-7.

Films with equal weight were taken and covered with 250 g of the soil in a transparent plastic box

The test were incubated at 37 C for 7 days, 14 days, 21 days and 28 days

Moisture of the soils were maintained by spraying water every days

Weight and photos of each samples will be taken after each time interval mentioned above

54
Photos of the experiment (2 hours curing time)

7 days 14 days 21 days

Samples for
the
experiments
28 days

55
Migration test of the films
Migration test of the films were carried out in four solvents namely water, ethanol, isooctane and acetic acid

Initial weight of the sample were taken and transferred into petri dish

10 mL of each of the solvent were poured into each plates

The sample with isooctane was incubated for 2 days at 20°C

Samples immersed in water, ethanol and acetic acid were incubated at 40°C for 10 dyas

After the finish of the experiments weight of each sample were taken to calculate the migration

Migration of the isooctane through the films were 62 mg cm2. (need to repeat)
Note : The solvent such as water, acetic acid and ethanol evaporated due to
melting of parafilm used

56
Study of temperature difference in the thermally insulated
aerogel based box

57
Wavenumber Functional groups
667 –OH stretching 1440 C-H bending vibration
830 P-OH 1626 deformation of water molecules
921 P-OH 2896 C-H stretching
1034 C-O-C stretching 3342 O-H vibration
1105 C-O stretching 1315 -CH2 in plane bending

1160 C-O-C asymmetric stretching of cellulose

58
Aerogel and bamboo stick house

59
Tensile strength of the threads (60 min curing time)

(a) (b)

Figure 1 (a) thread prepared without any solvent and (b) thread prepared in
isopropanol

60
Thread prepared in isopropanol curing time 30 min

61
Migration study of the films

Initial Final Migration Solvent Curing

weight of weight of (g/cm2) time
the films the films
(g)
0.0766 0.0735 0.0030 Isooctane 2 hrs
0.0563 0.0535 0.0028 Isooctane 1 hrs
0.1255 0.1212 0.0041 Isooctane 30 min
0.1077 0.0991 0.0086 Ethanol 1 hrs
0.0985 0.0892 0.0925 Ethanol 2 hrs

62
TGA analysis of the Films

63
 Continuous degradation and antibacterial activities of
CPM0@0.5%
Reactor (1.0 g CPMo@0.5%)
Peristaltic pump
(Flow rate 5 Methylene blue dye
mL/min) + Nutrient broth+
E.coli

Fig. 1 Experimental set up for continuous degradation and antibacterial activities

Fig. 1 Effects of CPMo@0.5% on the bacterial activities of E.coli during continuous

process
64
 Continuous degradation and antibacterial activities of
CPM0@0.5% in nutrient broth

Fig. 3 Represent the effect of CPMo@0.5% on E.coli (antibacterial activities without MB)

65
Effects of MB on the growth of E.coli

Fig. 4 Represents the effect of MB on the growth of E.coli

66
Absorption experiments of cellulose phosphate for diaper

Bamboo Bamboo sheet After water

cellulose Water
experimental set absorption absorption
phosphate sheet up (11 x 7 x 2.1
(11 cm x 7x 1.1 cm)
mm)

Initial weight of the cellulose phosphate paper = 3.63g

Final weight of the cellulose phosphate paper after water absorption = 70.15 g
Volume of water absorbed = 67 g
% water absorption = 1832.5%

67
Antibacterial properties of CPMo composites against gram
+ve bacteria (S. aureus)

Antibacterial activities of the cellulose phosphate MoS2 composites.

Bulk = Mo
0.1% = CPMo@0.1%, 0.25% = CPMo@0.25%, 0.5% = CPMo@0.5%,
0.75% = CPMo@0.75% , 1% = CPMo@1% and Control +ve = only bacteria

68
 Phosphorylation of bacterial cellulose

in Air dry
m
30
e
tim
g
rin
Cu

Phosphorylated Dried film like Tra

bacterial cellulose phosphorylated
in

bacterial cellulose
5 m
e 4
tim
ng

cellulose Freeze drying

ri
Cu

Phosphorylated Lyophilized film

bacterial cellulose like phosphorylated
bacterial cellulose

69
Biodegradation of cellulose phosphate film
Sample Days Degradation
percentage (%)

CP@120 min 7 days 51.53

14 days 57.30

CP@90 min 7 days 42.61

14 days 55.92

CP@ 60 min 7 days 26.98

14 days 28.45

21 days 31.30

70
Flame retardant test (Viscose)
FCT-60

T=0 s T=5 s T=10 s T=15 s T=30 s

71
 Flame retardant test (Bamboo)
PBF-30

T=0 s T=5 s T=10 s T=20 s T=40 s

PBF-60

T=0 s T=5 s T=10 s T=20 s T=40 s

PBF-60

T=0 s T=5 s T=10 s T=20 s T=40 s

72
Bamboo film Transparency

PBF-30 PBF-60

73
 Water absorption and swelling behaviour
PBF-30 (% ABSORPTION) PBF-60 (% ABSORPTION)
Initial weight Final weight Percentage Initial weight Final weight Percentage
(g) (g) absorption (g) (g) absorption
(%) (%)
0.0197 0.157 88.85 0.0266 0.453 94.13
PBF-30 (% swelling) PBF-60 (% swelling)

Initial height Final height % swelling Initial height Final height % swelling
(mm) (mm) (mm) (mm)
0.06 0.423 85.82 0.073 0.963 92.38

74
 Rice husk to silica preparation

1000 °C
2 hrs Holding time 5
s hr
hr s Yield %
5 25.5%

Delignified rice husk Silica

White White

400 °C 600 °C 600 °C

Holding time Holding time Holding time
2 hrs 2 hrs 2 hrs

White Black Grey Grey-white

Yield %
25%

75
Glass formation

1500 °C
5 hrs Holding time 9
s hr
hr s Yield %
9

Room temperature Room temperature

Silica
White

Experiment failed

76
XRD

77
Size distribution of the extracted silica

78
Phosphate content in Wheat straw film

266.6 mmol/kg
200 mmol/kg

350 mmol/kg

79
Wheat straw film FTIR

80
Mechanical properties of bamboo films

Sample Tensile Strain Tensile

strength (%) Modulus
(MPa) (MPa)
PBF-30 69.21 0.07 988.71
PBF-60 53.26 0.038 1404.2
PBF-90 51.42 0.04 1285.5

81
Water absorption property of wheat straw

Sample Avg. initial Avg. final % absorption

weight weight

WSF-30 0.0265 0.2423 814

WSF-60 0.03 0.164 447.5

WSF-75 0.07 0.3411 387.3

82
Phosphorylated wheat straw cup

83
P-content from phosphorylated bamboo

84
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LCA of cellulose phosphate gel production from wheat straw

85
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86
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87