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Rosli, N. A., Ahmad, I., and Abdullah, I. (2013). "Isolation and characterization of cellulose nanocrystals from Agave angustifolia fibre," BioRes. 8(2), 1893-1908.

Abstract

Cellulose nanocrystals were extracted from Agave angustifolia fibres by alkali and bleaching treatments followed by acid hydrolysis. The chemical composition of the Agave fibres was determined at different stages of chemical treatment. The structural analysis was carried out by a Fourier Transform Infrared spectroscopy and X-ray diffraction. The morphology and thermal stability of the Agave fibres at different stages of chemical treatment were investigated by field emission scanning electron microscopy and thermogravimetric analysis, respectively. The results indicated that the hemicellulose and lignin were removed extensively from the extracted cellulose. The two peaks at 1735 cm-1 and 1247 cm-1, which were attributed to the C=O stretching and C-O out of plane stretching vibration of the hemicellulose and lignin in raw Agave, completely disappeared in the spectra of chemically treated fibres. The X-ray diffraction data showed enrichment in the portion of crystalline cellulose from 59% to 82% in the raw and cellulose nanocrystals, respectively. Thermogravimetric analysis showed that the thermal stability improved significantly by various chemical stages. The size reduction of the Agave cellulose into nano-sized particles from 7 µm to 8 nm in diameter by acid hydrolysis was confirmed with transmission electron microscopy images.


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Isolation and Characterization of Cellulose Nanocrystals from Agave angustifolia Fibre

Noor Afizah Rosli, Ishak Ahmad,* and Ibrahim Abdullah

Cellulose nanocrystals were extracted from Agave angustifolia fibres by alkali and bleaching treatments followed by acid hydrolysis. The chemical composition of the Agave fibres was determined at different stages of chemical treatment. The structural analysis was carried out by a Fourier Transform Infrared spectroscopy and X-ray diffraction. The morphology and thermal stability of the Agave fibres at different stages of chemical treatment were investigated by field emission scanning electron microscopy and thermogravimetric analysis, respectively. The results indicated that the hemicellulose and lignin were removed extensively from the extracted cellulose. The two peaks at 1735 cm-1 and 1247 cm-1, which were attributed to the C=O stretching and C-O out of plane stretching vibration of the hemicellulose and lignin in raw Agave, completely disappeared in the spectra of chemically treated fibres. The X-ray diffraction data showed enrichment in the portion of crystalline cellulose from 59% to 82% in the raw and cellulose nanocrystals, respectively. Thermogravimetric analysis showed that the thermal stability improved significantly by various chemical stages. The size reduction of the Agave cellulose into nano-sized particles from 7 µm to 8 nm in diameter by acid hydrolysis was confirmed with transmission electron microscopy images.

Keywords: Acid hydrolysis; Cellulose; Nanocrystals; Natural fibres

Contact information: Polymer Research Centre (PORCE), School of Chemical Sciences and Food Technology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi Selangor, Malaysia; *Corresponding author: gading@ukm.my

INTRODUCTION

Biocomposites processing using natural fibres as reinforcement has increased dramatically in recent years (Jiang and Hinrichsen 1999; Luo and Netravali 1999; Takagi and Asano 2008; Takagi and Ichihara 2004). In addition, natural fibres have found extensive applications in textiles, paper manufacturing, building, and civil engineering fields (Kalia et al. 2009).

Natural fibres of lignocellulosic materials can be classified according to their plant origin i.e.: (i) bast or stem, (ii) leaf, (iii) seed or fruit, (iv) grass, and (v) straw fibres. Natural fibres have many advantages over man-made fibres, such as low density, low cost, availability, renewability, biodegradability, and low abrasivity (Bledzki et al.1996). A better understanding of the chemical, mechanical, and physical properties of natural fibres is necessary for processing natural fibre-reinforced composites. Natural fibres consist of cellulose, hemicelluloses, lignin, pectin, waxes, and water soluble substances. Cellulose is a semi-crystalline polysaccharide consisting of D-glucopyranosyl units linked together by β-(1-4)-glucosidic bonds (Bledzki and Gassan 1999). Three hydroxyl groups, at the C2 and C3 positions of secondary hydroxyl groups, and the C6 position of primary hydroxyl groups, can form intra- and intermolecular hydrogen bonds. These hydrogen bonds allow the creation of highly ordered, three dimensional crystal structures (Abdul Khalil et al.2011). Lignin is a polymer of phenylpropane units which are highly complex, mainly aromatic and amorphous, but have less water sorption than other natural fibre components (Rowell et al. 1997). Hemicellulose is branched, fully amorphous, and has a lower molecular weight than cellulose; as a consequence it is partly soluble in water. Hemicellulose is hygroscopic due to its open structure containing hydroxyl and acetyl groups (Frederick and Norman 2004).

The purpose of this study was to produce cellulose nanocrystals (CNCs) from Agave angustifolia as a substitute for synthetic fibres to make a good economic and ecologic composite material. The term “cellulose nanocrystal” is used to designate elongated crystalline rod-like cellulose nanoparticles. Agave angustifolia belongs to the Agavaceae family and is one of 140 species of the Agave genus. Agave angustifolia is known as the “Century Plant” or locally named as “Kelumpang Telur”. The leaf of A. angustifolia is about 1½ inches wide with creamy yellow stripes along the spiny margins as shown in Fig. 1. The plant grows into a spherical clump of 3 to 4 feet in diameter. This species can tolerate full sun, part shade, and reflected heat. It can also handle more water than most Agave species. It has been utilized by people for domestic purposes such as making rope, soap, and other products. Although the leaves are widely used, no studies on the extraction or properties of the cellulose fibres from these leaves have been conducted to date.

Fig. 1. Photograph of Agave angustifolia plants

CNCs have great potential as reinforcing agents in nanocomposites due to their size and the possibility of chemically modifying their surface. It is known that the conventional CNC exhibits high stiffness and modulus, which can be as high as 134 GPa (Oksmann and Sain 2006). It has been stated that the crystal structure exhibits a tensile strength from 0.8 up to 10 GPa (Nishino et al. 1995).

A well known process for the isolation of CNCs is strong acid hydrolysis. It allows the removal of the form crystalline nanoparticles. Many researchers have recently used this method to prepare CNCs, from kenaf (Kargarzadeh et al. 2012), banana plant (Elanthikkal et al. 2010), coconut husk (Rosa et al. 2010), rice husk (Johar et al. 2012), and sugarcane bagasse (Mandal and Chakrabarty 2011).

In this study, cellulose and CNCs were extracted from Agave fibres by chemical methods of alkali and bleaching treatments followed by acid hydrolysis. The next study aims for the application of cellulose and CNCs of Agave fibre in biocomposites. Thus, investigations on the chemical and physical properties were carried out to analyse their suitability as reinforcing agent in biocomposites.

EXPERIMENTAL

Materials

Agave leaves used in this study were harvested in Kajang, Selangor (Malaysia). The chemical reagents used were sodium chlorite (NaClO2, Sigma-Aldrich), acetic acid glacial (99%, Systerm), sodium hydroxide (NaOH, Systerm), sulphuric acid (H2SO4, 98%, Univar), methanol (R&M Chemicals), toluene (Systerm), and acetone (Systerm).

Extraction of Cellulose

The thorns on the mature Agave leaf margin and tips were removed. The leaves were dried for three days to remove excess moisture. Agave fibres were extracted from the leaves using fibre extracting machine and were retted for one week. Then the fibres were combed and shadow-dried for 4 h. The long fibres were cut to 3 to 5 cm and ground using a mill. The ground fibres were treated with 4% NaOH at 70 to 80 ºC for 2 h, after which bleaching treatment was carried out using 1.7 w/v% NaClO2 at 70 to 80 ºC for 4 h. The ratio of the fibres to liquor was 1:25 (g/mL). Each fibre treatment was done twice, and the fibres were washed with distilled water after each treatment.

Isolation of Cellulose Nanocrystals

Scheme 1 depicts the methodology routes to obtained CNCs from Agave fibres. The hydrolysis was carried out using 60 wt% H2SO4solutions at 45 ºC for 45 min. The ratio of the obtained cellulose to liquor was 1:20 (wt%).

Scheme 1. Scheme for CNCs production from Agave fibre

The hydrolyzed cellulose was washed five times by centrifugation (10,000 rpm, 10 min, and 10 ºC). After washing, the products were neutralized with 2 N NaOH to a pH of 7. The neutralized products were further washed three more times. The suspension was then dialyzed against distilled water to a constant pH.

Determination of Chemical Composition

The chemical composition of the Agave fibres before and after the chemical treatment was determined. Dewaxing was carried out by boiling the fibres in a mixture of toluene/methanol (1:1 v/v) in a sachet for 6 h. The de-waxed fibres were then filtered, washed with methanol, and dried. The percentage of holocellulose was calculated according to the method described previously (Wise et al. 1946). In this method, the extracted residue of de-waxed fibres was boiled in a mixture of NaClOand acetic acid for 4 h. The suspension was cooled in an ice bath for 30 min, filtered, and washed with cold distilled water. Finally, it was washed with acetone and dried. The α-cellulose and the acid-insoluble lignin content was determined according to TAPPI standard methods T203 and T222, respectively.

Field Emission Scanning Electron Microscopy

Field emission scanning electron (FESEM) micrographs of the treated and untreated fibres were recorded on a Zeiss Supra 55VP microscope at a voltage of 3 kV. The micrographs of the longitudinal and cross section of the Agave leaves were also recorded. The samples were coated with gold to avoid charging.

Transmission Electron Microscopy

In order to determine the dimensions of the CNCs, the hydrolyzed suspension were analyzed by a Philips CM12 transmission electron microscope (TEM) operating at 80 kV. A drop of a highly diluted suspension of CNCs was placed on a copper grid coated with a thin carbon film and allowed to dry at room temperature. The copper grid was then stained with a 2 wt% solution of uranyl acetate for one minute and air dried.

Fourier Transform Infrared Spectroscopy

The Fourier transform infrared (FTIR) spectra of the Agave fibres were recorded on a Perkin-Elmer spectrometer (SpectrumGX) in the range 500 to 4000 cm-1 with a scanning resolution of 8 cm-1. Before analysis, all of the samples were ground into a fine powder and dried for 24 h at 60 ºC in an oven. The ground fibres was mixed with KBr, and pressed into an ultra-thin pellet.

X-ray Diffraction

X-ray diffraction (XRD) analysis was performed with a Bruker AXS D8 Advance diffractometer at 40 kV, 40 mA with Cu-Kα radiation (λ=0.1541 nm). Before analysis, the samples were ground into a fine powder using a mill and pressed into a sample holder. The data was acquired in a 2θ range from 5 to 60º. The crystallinity index (lc) of Agave fibres at different chemical stages was calculated using the following formula (Segal et al. 1959).

(1)

where l002 is maximum intensity of diffraction of the (002) lattice peak (22º to 23º), and lam is that of the amorphous material between 18º to 19º where the intensity is minimum.

Thermogravimetric Analysis

A Mettler Toledo model TGA/SDTA851e thermogravimetric analyzer was used to characterize the thermal stability of the Agave fibres. Approximately 2 mg of each sample was placed in an aluminium pan and heated from 30 to 600 ºC at a heating rate of 10 ºC/min. All of the measurements were performed under a nitrogen atmosphere.

RESULT AND DISCUSSION

Chemical Composition

The chemical compositions of raw, alkali treated, and bleached Agavefibres are presented in Table 1. The raw Agave fibres are composed of 67.0% cellulose, 25.2% hemicelluloses, 6.3% lignin, and 2.5% extractives. The amount of hemicelluloses and lignin are higher in raw fibre as compared to the treated fibres. NaOH was found to be efficient in removing the hemicellulose from the fibre, as hemicellulose content was decreased from 25.2% to 3.9%. Based on the chemical composition analysis, most of the lignin content was removed by the bleaching treatment, in which it reacts with NaClO2to dissolve as a lignin chloride. Bleaching does not only remove lignin, but also some of the hemicellulose. The final fibre obtained after the bleaching treatment was found to have the highest cellulose content (97.3%).

Table 1. Chemical Composition of Agave Fibres at Different Stages of Chemical Treatment

Morphological Analysis

Figure 2 shows the Agave angustifolia fibres at different stages of the chemical treatment. The colour of the fibres changed from cream to light brown after the alkali treatment, and white after bleaching.