Synthesis, Optimization, and Characterization of Cellulose

Nanocrystals from Sugarcane Bagasse using Acid Hydrolysis

1. Introduction
1.1 Back Ground of the Study

In recent years, the quest for sustainable development has motivated efforts toward maximizing
the efficiency of the use of raw materials and minimizing the creation of waste (Ashori, 2008).
The reuse of these residues allows a significant reduction both in the volume of waste
accumulated in the environment and in the extraction of raw materials for various applications.
Thus, an efficient reuse of these wastes is of great importance, not only for minimizing the
environmental impact, but also for obtaining a higher profit (Ghazy et al., 2016). Sugarcane
bagasse (SCB) is a residue from the refining process of sugarcane that contains about 40-50% of
cellulose in its composition (Ghazy et al., 2016; Sun et al., 2004). In Ethiopia, sugarcane is
cultivated on about 33,777 ha of the country’s land, located around the Wonji-Shoa, Metahara
and Finchaa sugar factories (Tilahun et al., 2014).The bagasse (fibrous portion) is burned to
provide heat and electricity for sugar mills and green tops can be used as cattle feed (Tilahun et
al., 2014). SCB mainly contains cellulose, lignin, hemicelluloses and other with small quantity
such as ash, silica and ethanol–toluene extractive substances (Andrade & Colodette, 2014).

Lignocellulose is the primary building block of plant cell walls. The complex hierarchy structure
of lignocellulosic biomass is the main obstacle for key components fractionation, where
cellulose, hemicellulose, and lignin are hindered by many physicochemical, structural, and
compositional factors (Lee et al., 2014).

1.2 Statement of the problem

Sugar factories are one of the agro- industries that use sugarcane as the main raw material and
generate huge amounts of wastes worldwide. Brazil is the world’s largest producer of sugarcane.
Likewise, Ethiopia is on the way to have about 10 sugar factories within a few years. These sugar
factories will generate huge amounts of sugarcane bagasse. Hence, a practical way of waste
management to use sugarcane bagasse is to understand existing technologies and integrate them
with further research for a use full application. The waste sugarcane bagasse obtained from sugar
industries may be a potential opportunity in the future to be used as a suitable lignocellulosic
feedstock for value-added materials.

It will be more worthwhile to generate cellulose nanocrystals from the wastes of sugar factories
with respect to the economic and ecological benefits. Some researchers have produced nano
sized cellulose crystals from sugarcane bagasse by acid hydrolysis using hydrochloric acid.
However, hydrolysis with hydrochloric acid results in reduced dispersion of crystals, leading to
their flocculation when in solution. In this study, sugarcane bagasse will be examined as a
lignocellulosic material source to produce cellulose nanocrystals due to its high contents of
cellulose, impressive growth rate with demand, and low competition on land use. This work aims
to produce cellulose nanocrystals from sugarcane bagasse using sulfuric acid as a hydrolyzing
agent.

1.3 Objective
1.3.1 General objective

To synthesis and characterize cellulose nanocrystals (CNCs) from sugarcane bagasse (SCB)
using acid hydrolysis method.

1.3.2 Specific objective

a. To extract and purify cellulose from sugarcane bagasse using appropriate solvents.
b. To synthesis cellulose nanocrystals from sugarcane bagasse using acid hydrolysis.
c. To investigate the effects of acid concentration, reaction temperature and hydrolysis time,
and to determine the optimum parameter on the yield of CNCs.
d. To characterize the physicochemical properties of synthesized CNCs.

2. Methods
2.1 Synthesis of Cellulose

Sugarcane bagasse will be collected from Wonji/Shoa sugar factory. To isolate and purify
cellulose from sugarcane bagasse, the raw sample will pass through different pretreatment steps.
The basic pretreatments such as size reduction, extraction of soluble components, bleaching and
alkaline treatment will be performed in this study for the removal of impurities and sample
homogenization. After size reduction the sample will be dewaxed in a Soxhlet apparatus using
mixture of toluene and ethanol solvents with the ratio of 2:1 (v/v). The de-waxing process will be
conducted at 85 oC for 5 h to remove extractive components such as wax, pectin and oils. The de
waxed powder washed with distilled water and ethanol repeatedly; filtered using 45 µm sieves to
remove solvents and finally dried in oven at 45 oC. Bleaching and alkaline pre-treatments are
also aid in the removal of surface waxes and extractives (Ghazy et al., 2016; Malucelli et al.,
2019). In bleaching process two treatments, namely treatment with chlorine compounds under
acidic condition (more effective), and treatment with hydrogen peroxide (H2O2) under alkaline
condition are well known (Malucelli et al., 2019). The acid chlorite treatment is widely used to
remove most of lignin and other components by the combination of distilled water, sodium
chlorite, and acetic acid stirring with lignocellulosic biomass at 70–80 oC for 4–12 h. The acetic
acid and sodium chlorite are fed to the mixtures at interval time, i.e., at every hour, for
controlling the pH value. The obtained solid products are collected and dried in oven at 50 oC,
which is defined as holocellulose, mainly including hemicellulose and cellulose in the fibers. The
white color fiber of holocellulose indicates the successful removal of lignin and other impurities.

2.2 Preparation of cellulose nanocrystals

Controlled H2SO4 hydrolysis will be performed in an autoclave reactor to split the amorphous
domains, remove local inter fibril crystalline contacts of the pretreated sample. Purified cellulose
is acid hydrolyzed using 10g of cellulose in 250 ml of sulfuric acid for one run (1g: 25ml) under
controlled conditions and continuous agitation at 500 rpm (Wulandari et al., 2016). The
hydrolysis reaction will be quenched by adding ice to the autoclave reactor. The hydrolyzed
cellulose sample will then be washed repeatedly with successive centrifugation at 4000 rpm for
25 min to remove excess sulfuric acids (Ghazy et al., 2016). The suspension is collected and
diluted with distilled water. The suspension will then be dialyzed against deionized water using
cellulose membrane for 4 days to neutralize and fully remove free acid molecules such as non-
reactive sulfate group, salts as well as soluble sugars. After dialysis, the obtained suspension of
CNCs will be ultrasonicated for 15 min to get aqueous homogenized dispersion and labeled as
the CNCs suspension. Then, a few drops of chloroform will be added to the freshly prepared
suspension to prevent degradation of the cellulose nanocrystals and then stored in a refrigerator
at 4oC for characterization.