Cellulose (2009) 16:10171023 DOI 10.

1007/s10570-009-9334-9

Comparison of the characteristics of cellulose microbril aggregates of wood, rice straw and potato tuber
Kentaro Abe Hiroyuki Yano

Received: 1 April 2009 / Accepted: 4 June 2009 / Published online: 23 June 2009 Springer Science+Business Media B.V. 2009

Abstract The focus of this study has been to isolate cellulose microbril aggregates by the one-time grinding treatment from wood, rice straw and potato tuber, and to compare their morphological and mechanical properties. Field emission scanning electron microscopy images showed that the diameter range of isolated microbril aggregates from wood, 1220 nm, was smaller than those from rice straw and potato tuber, 1235 nm and 1255 nm, respectively. These differences were observed even in the puried rice straws and potato tuber before the grinder treatment, but were hardly observed in the puried wood. The results of X-ray analysis and tensile tests indicated that there were no signicant differences among the sources in the cellulose crystallinity and Youngs modulus of the isolated microbril aggregates in the dry state. These results suggest that the inherent characteristics of cellulose microbril aggregates in the dry state are very similar regardless of plant sources and tissue functions. Keywords Cellulose microbril Crystallinity Douglas r Potato tuber Rice straw Tensile property

Introduction Social concerns for sustainable green products are encouraging the efcient exploitation of cellulose, the most abundant renewable biopolymer on earth. Cellulosic bers traditionally have been used to make paper and clothing because of their high stiffness, low density and low cost, and recently they have entered the spotlight as nanomaterials. Cellulose exists in plant cell walls as highly crystalline microbrils with several nanometers width that are formed when hydrogen bonding causes long cellulose molecules to become laterally packed. Thus, cellulose microbrils exhibit outstanding mechanical properties, such as a high Youngs modulus (138 GPa) in the crystal region along the longitudinal direction (Sakurada et al. 1962) and a very low coefcient of thermal expansion (10-7 K-1) along the longitudinal direction (Nishino et al. 2004), which make them promising candidates for reinforcement materials in nanocomposites. Many researchers are now tackling the isolation of nanosized cellulose whiskers and bers from plant sources along with their efforts to utilize the nano elements (Dufresne et al. 1997; Dinand et al. 1999; Dufresne and Vignon 1998; Zhao et al. 2007; Abe et al. 2007; Zuluaga et al. 2007; Alemdar and Sain 2008). In particular, nanobers, consisting of either single cellulose microbrils or microbril aggregates, exhibit high reinforcing efciency because of their high aspect ratios (Nakagaito and Yano 2004; Zimmerman et al. 2005; Iwamoto et al. 2005).

K. Abe (&) H. Yano Research Institute for Sustainable Humanosphere, Kyoto University, Gokasho, Uji, Kyoto 611-0011, Japan e-mail: abekentaro@rish.kyoto-u.ac.jp

123

1018

Cellulose (2009) 16:10171023

The cellulose nanober sources are as diverse such as wood and many agricultural residues and so far they have been isolated and characterized from wood (Zhao et al. 2007; Abe et al. 2007), wheat straw and soy hulls (Alemdar and Sain 2008), banana rachis (Zuluaga et al. 2007), sugar beet (Dufresne et al. 1997; Dinand et al. 1999) and potato tuber (Dufresne and Vignon 1998), and so on. Naturally, these plant sources differ in terms of tissue structure or function. For example, wood consists mostly of secondary xylem, and wheat straw is a complex of vascular bundles, parenchyma and epidermis. Sugar beet and potato tuber are composed mostly of parenchyma cells. Therefore, there is great interest from a material viewpoint to compare the morphology and mechanical performance of cellulose nanobers isolated from different plant sources. However, the use of different isolation methods in the above various reports makes it difcult to impartially compare isolated nanobers. Actually, it is a laborious task to isolate ne nanofibers from multilayered structures such as wood bers. Still more difcult is the isolation of nanobers when hydrogen bonds are formed between microbrils, like in dried pulp (Iwamoto et al. 2008). Hence, repeated mechanical treatment using a high-pressure homogenizer or microuidizer is often required to extract nanobers (Dufresne et al. 1997; Dufresne and Vignon 1998; Dinand et al. 1999; Leitner et al. 2007; Paakko et al. 2007; Zuluaga et al. 2007). However, this excessive treatment reduces cellulose crystallinity and the degree of polymerization, leading to the degradation of mechanical properties of isolated nanobers (Iwamoto et al. 2007). Consequently, the potential properties of cellulose nanobers isolated from different plant sources remain to be claried. In a previous paper, we reported an efcient method for extracting cellulose nanobers from wood using a one-time grinding treatment of undried wood after the removal of matrix substances such as hemicelluloses and lignin (Abe et al. 2007). The isolated nanobers had a uniform width of approximately 15 nm and a length of more than several micrometers and corresponds to the cellulose microbril aggregates that exist in wood cell walls (Donaldson 2007). Furthermore, the mechanical treatment is slight enough to avoid the degradation of bers, implying that the nanobers maintain similar physical and mechanical characteristics of the microbril aggregates in wood cell walls. In this

study, we applied the one-time grinding treatment to plant sources other than wood, such as rice straw, which is a herbaceous plant, and potato tuber pulp, which consists mostly of parenchyma cells, and then compared the morphology and mechanical properties of the isolated cellulose nanobers with those of wood nanobers. Rice straw and potato pulp are major agricultural by-products and are renewable sources with great potential.

Materials Softwood, rice straw and potato tuber pulp were chosen as plant sources for the preparation and analysis of cellulose nanobers. Wood powder from Douglas r (Pseudotsuga menziesii [Mirb.] Franco) sieved under 60 mesh was used. The rice (Oryza sativa L.) straw was collected from local farms in Japan in 2007, and only stems cut into 3 cm lengths were used. Potato pulp powder, the by-product of the removal of starch granules from potato (Solanum tuberosum L.) tuber, was made in Japans Hokkaido prefecture. All samples were air-dried and stored at room temperature.

Methods Preparation of cellulose nanober The isolation of cellulose nanobers from the three sources was performed as follows, referring to our previous experiments (Abe et al. 2007). The samples were puried by a series of chemical treatments and then were subjected to a mechanical treatment according to the owchart shown in Fig. 1. For wood and rice straw, the samples were rst dewaxed in a Soxhlet apparatus with a 2:1 (v/v) mixture of toluene/ ethanol for 6 h. Afterward, a sodium chlorite (NaClO2) treatment, following the method of Wise et al. (1946), mainly removed lignin, and an alkaline treatment with potassium hydroxide (KOH) removed hemicelluloses, residual starch and pectin. After the series of chemical treatments, the samples were ltered and rinsed with distilled water until the residues were neutralized. The water slurry with 1 wt% undried puried sample was passed once

123

Cellulose (2009) 16:10171023

1019

X-ray diffraction The puried samples and the isolated nanobers were subjected to X-ray diffraction measurement by the transmission method. The puried samples dried at 105 C were made into pellets by pressing. For the isolated nanobers, water slurry at a ber content of 0.2 wt% was prepared, then converted by ltration into thin sheets 80 lm in thickness and dried at 55 C. In order to obtain enough intensity, eightlayered sheets were used for the measurement. Equatorial diffraction proles were obtained with Ni-ltered CuKa (k = 0.154 nm) from an X-ray generator (UltraX 18HF; Rigaku Corp., Tokyo, Japan) operating at 30 kV and 100 mA. The diffraction prole was detected using an X-ray goniometer scanning from 5 to 40. Five samples for each were subjected to the measurement. The relative degree of cellulose crystallinity was calculated according to the Segal method (Segal et al. 1959). The average values of cellulose crystallinity were calculated from the ve samples. Tensile test
Fig. 1 Experimental procedure for the preparation of cellulose nanobers from each plant source

through a grinder (MKCA6-3; Masuko Sangyo Co., Ltd., Saitama, Japan) at 1,500 rpm. The grinding treatment was performed with the clearance gauge of -6 (corresponding to 0.6 mm shift) from zero position, which was determined by the point of slight contact between the grinding stones. Field-emission scanning electron microscope (FE-SEM) The nanober slurries were diluted with 10 times the volume of ethanol, and then the cast lms were prepared at 105 C. The lm was coated with platinum by an ion sputter coater and was observed with a FE-SEM (JSM-6700F; JEOL Ltd., Tokyo, Japan) operating at 1.5 kV. Although the coating thickness was approximately 2 nm in this condition, we conrmed that the coating did not change the lateral dimension of brils signicantly. The diameter range of isolated nanobers was estimated from the diameter of 30 nanobers obtained by manual measurement.

Thin sheets prepared for the X-ray diffraction measurement were subjected to a tensile test using a universal materials testing machine (model 3365; Instron Corp., Canton, MA) at a crosshead speed of 1 mm/min with a gauge length of 10 mm. The load cell capacity was 5 kN. The dimensions of the sheet strips were 20 mm in length by 3 mm in width by 80 lm in thickness. The specic modulus and the specic strength were calculated by dividing the experimental values of the Youngs modulus and the tensile strength by the bulk density of the sheet, respectively. The density was calculated by measuring the average thickness at nine points and air-dried weight. The average values of the Youngs modulus and the tensile strength were calculated from the ve samples.

Results FE-SEM observations The viscosities of the slurries from wood, rice straw and potato tuber pulp were similarly high after the

123

1020

Cellulose (2009) 16:10171023

one-time grinding treatment, and none of the slurries showed macroscopically visible ber. Figure 2 compares FE-SEM images of the isolated bers from the three sources. No bers larger than about 60 nm in width were found in any of the FE-SEM images, conrming that ne nanobers were successfully obtained from rice straw and potato tuber as effectually as from wood. In higher plants, the diameter of a single microbril is reported to be *3 nm (Somerville et al. 2004). Thus, the uniform nanobers isolated in this study are so-called microbril aggregates (Donaldson 2007). Interestingly, the range of diameter distribution varied among the sources. That is, the diameter range of isolated microbril aggregates from wood, 1220 nm, was smaller than those from rice straw and potato tuber, 1235 nm and 12 55 nm, respectively. One may think that the greater thickness of the nanobers obtained from rice straw and potato tuber is an experimental artifact attributable to aggregations due to the removal of hemicelluloses (McCann et al. 1990) and drying for the cast lms. However, thicker aggregates were observed even in the puried rice straws and potato tuber (Fig. 2e, f) before the grinder treatment, but were hardly observed in the puried wood (Fig. 2d). Thus, it is plausible that the patterns of aggregation of cellulose microbril differ among plant sources and cell types.

X-ray analysis Cellulose crystallinity in isolated nanobers is of interest as the key factor determining their mechanical properties. Figure 3 and Table 1 show the equatorial X-ray diffraction patterns and the relative degrees of cellulose crystallinity, respectively, of the puried samples and the isolated nanobers. The patterns of all of the puried samples were the typical Cellulose I pattern and were nearly identical. In fact, in relative crystallinity there was no signicant difference among the sources, as shown in Table 1. Besides, when the a-cellulose content of the puried samples was determined by extraction with 17.5% NaOH, the a-cellulose contents were almost the same among the sources, approximately 85%. These results suggest that the intrinsic crystallinities of cellulose microbrils in wood, rice straw and potato tuber were equivalent before isolation regardless of the type of cell. Although the detailed observation of the diffraction patterns of all the samples sometimes indicated small differences in the half-width and in the position of the diffraction peaks, no signicant differences were found in the corresponding mean values. Furthermore, the isolated nanobers from the three sources had almost identical diffraction patterns and relative crystallinities. Considering that ne

Fig. 2 FE-SEM micrographs of the isolated cellulose nanobers from a wood, b rice straw, c potato tuber and microbril aggregates in a puried samples before isolation from d wood, e rice straw, f potato tuber. Arrows indicate thicker aggregates

123

Cellulose (2009) 16:10171023

1021

a one-time grinder treatment was performed on all sources in this study, it is likely that the degree of mechanical degradation was minimal. Therefore, this increment, which can be explained by the removal of the amorphous cellulose and hemicelluloses during the ltration of the nanobers for sheet preparation, implies that the grinding treatment causes little damage to the crystal regions of the nanobers. Tensile test The FE-SEM observations and X-ray analysis showed that ne cellulose nanobers were isolated with minimal damage from rice straw and potato tuber as effectually as from wood. Hence, the next subject of interest would be the mechanical properties of the nanobers derived from such different sources. However, it is quite difcult to directly measure the tensile properties of isolated nanobers. Thus, we compared the mechanical properties of sheets prepared from the nanobers (Table 2). To eliminate the property variation due to the sheet density, the mechanical properties were compared by specic values: the specic modulus and specic tensile strength. These comparisons showed no signicant difference among the three sources. Because the results of tensile tests, especially tensile strength, depend on the hydrogen bonding between bers, the results do not necessarily mean that the individual cellulose nanobers from the three sources have similar tensile strength. However, the nanober sheets having similar densities and cellulose crystallinities and similar FE-SEM images regardless of the source imply that the bonding patterns of the nanober sheets of the three sources were not signicantly different. Likewise the cellulose nanobers in the dry state have similar tensile properties, at least in Youngs modulus, among all three sources. Considering that those nanobers are microbril aggregates extracted from the cell walls with little damage, as proven by the X-ray analysis, it seems that the mechanical properties of microbril aggregates from wood, rice straw and potato tuber in the dry state are similar.

Fig. 3 Equatorial X-ray diffraction proles of a puried samples and b isolated nanobers from wood (black), rice straw (gray) and potato tuber (white)

Table 1 Average relative degrees of cellulose crystallinity of puried samples and isolated nanober from different plant sources Source Relative crystallinity (%) Puried sample Wood Rice straw Potato tuber 71 (1) 68.2 (0.9) 66.1 (0.6) Isolated nanober 78.6 (0.7) 76.0 (0.8) 80.2 (0.8)

Note: Standard deviations are given in parentheses (n = 5)

nanobers were obtained from all of the sources, as shown in FE-SEM images, it can be said that the structural differences of the cells do not affect the easiness of nanober isolation when a grinder is used. It is also interesting that the isolated nanobers exhibit higher crystallinity than the puried samples regardless of the source. Iwamoto et al. (2007) reported the decrease of the degree of polymerization of the isolated cellulose nanobers as the number of passes through the grinder increased. However, because only

Discussion In this study, cellulose nanobers (microbril aggregates) were isolated from wood, rice straw and potato

123

1022 Table 2 Average mechanical properties of nanober sheets from different plant sources Source Wood Rice straw

Cellulose (2009) 16:10171023

Density (g/cm3) Youngs modulus (Gpa) Specic modulus (GPa) Tensile strength (MPa) Specic strength (MPa) 1.31 1.36 11 (2) 11 (1) 11.4 (0.6) 8 (1) 8 (1) 8.3 (0.4) 210 (20) 230 (30) 230 (10) 160 (20) 170 (20) 168 (9)

Potato tuber 1.34

Note: Standard deviations are given in parentheses (n = 5)

tuber, and their morphological and mechanical properties were compared. The one-time grinding treatment in an undried state after the removal of the matrix substances enabled the isolation of ne nanobers from all three sources. FE-SEM observations showed that the diameter range of isolated nanobers from wood was slightly smaller than those from rice straw and potato tuber. However, there were no signicant differences among the sources in the relative cellulose crystallinity of the isolated nanobers. Furthermore, the tensile tests of nanober sheets obtained from all three sources exhibited similar properties. These results imply that the choice of plant source does not limit the range of application of the isolated nanobers. In the softwood sources used in this study, the tissue was composed of 9095% longitudinal tracheids, whose functions are mainly structural support and water conduction. On the other hand, the main function of parenchyma cells, which occupy most of the tissue of potato tuber, is the storage of food and water. This study suggested that the inherent characteristics of cellulose microbril aggregates in the dry state are very similar regardless of plant sources and tissue functions. However, this does not mean that the cellulose microbrils existing in the cell walls of tracheid and parenchyma have similar cellulose crystallinity and mechanical properties. It is quite possible that the characteristics of the microbrils synthesized are controlled by environmental conditions such as the chemical composition of the polysaccharides and moisture content. Further analysis of the characteristics of cellulose nanobers or cellulose microbrils from many kinds of plants will be necessary for the efcient exploitation of cellulose, the most abundant biopolymer on earth.
Acknowledgments This work was supported by Grant-inAid from Research Fellowships of the Japan Society for the Promotion of Science for Young Scientists.

References
Abe K, Iwamoto S, Yano H (2007) Obtaining cellulose nanobers with a uniform width of 15 nm from wood. Biomacromolecules 8:32763278 Alemdar A, Sain M (2008) Isolation and characterization of nanobers from agricultural residues: wheat straw and soy hulls. Bioresour Technol 99:16641671 Dinand E, Chanzy H, Vignon MR (1999) Suspensions of cellulose microbrils from sugar beet pulp. Food Hydrocoll 13:275283 Donaldson L (2007) Cellulose microbril aggregates and their size variation with cell wall type. Wood Sci Technol 41:443460 Dufresne A, Vignon MR (1998) Improvement of starch lm performances using cellulose microbrils. Macromolecules 31:26932696 Dufresne A, Cavaille JY, Vignon MR (1997) Mechanical behavior of sheets prepared from sugar beet cellulose microbrils. J Appl Polym Sci 64:11851194 Iwamoto S, Nakagaito AN, Yano H, Nogi M (2005) Optically transparent composites reinforced with plant ber-based nanobers. Appl Phys A 81:11091112 Iwamoto S, Nakagaito AN, Yano H (2007) Nano-brillation of pulp bers for the processing of transparent nanocomposites. Appl Phys A 89:461466 Iwamoto S, Abe K, Yano H (2008) The effect of hemicelluloses on wood pulp nanobrillation and nanober network characteristics. Biomacromolecules 9:10221026 Leitner J, Hinterstoisser B, Wastyn M, Keckes J, Gindl W (2007) Sugar beet cellulose nanobril-reinforced composites. Cellulose 14:1925 McCann MC, Wells B, Roberts K (1990) Direct visualization of cross-links in the primary plant cell wall. J Cell Sci 96:323334 Nakagaito AN, Yano H (2004) The effect of morphological changes from pulp ber towards nano-scale brillated cellulose on the mechanical properties of high-strength plant ber based composites. Appl Phys A 78:547552 Nishino T, Matsuda I, Hirao K (2004) All-cellulose composite. Macromolecules 37:76837687 Paakko M, Ankerfors M, Kosonen H, Nykanen A, Ahola S, Osterberg M, Ruokolainen J, Laine J, Larsson PT, Ikkala O, Lindstrom T (2007) Enzymatic hydrolysis combined with mechanical shearing and high-pressure homogenization for nanoscale cellulose brils and strong gels. Biomacromolecules 8:19341941 Sakurada I, Nukushima Y, Ito I (1962) Experimental determination of the elastic modulus of crystalline regions in oriented polymers. J Polym Sci 57:651660

123

Cellulose (2009) 16:10171023 Segal L, Creely JJ, Martin AE, Conrad CM (1959) An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer. Text Res J 29:786794 Somerville C, Bauer S, Brininstool G, Facette M, Hamann T, Milne J, Osborne E, Paredez A, Persson S, Raab T, Vorwerk S, Youngs H (2004) Toward a systems approach to understanding plant cell walls. Science 306:22062211 Wise LE, Murphy M, DAddieco AA (1946) Chlorite holocellulose, its fractionation and bearing on summative wood analysis and on studies on the hemicelluloses. Pap Trade J 122:3543

1023 Zhao HP, Feng XQ, Gao H (2007) Ultrasonic technique for extracting nanobers from nature materials. Appl Phys Lett 90:073112 Zimmerman T, Pohler E, Schwaller P (2005) Mechanical and morphological properties of cellulose bril reinforced nanocomposite. Adv Eng Mater 7:11561161 Zuluaga R, Putaux JL, Restrepo A, Mondragon I, Ganan P (2007) Cellulose microbrils from banana farming residues: isolation and characterization. Cellulose 14:585 592

123