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BIOMAC 5067 1–7 ARTICLE IN PRESS

International Journal of Biological Macromolecules xxx (2015) xxx–xxx

Contents lists available at ScienceDirect

International Journal of Biological Macromolecules

journal homepage: www.elsevier.com/locate/ijbiomac

1 Preparation of food grade carboxymethyl cellulose from corn husk

2 agrowaste
3 Q1 Md. Ibrahim H. Mondal ∗ , Mst. Sarmina Yeasmin, Md. Saifur Rahman
4 Polymer and Textile Research Lab., Department of Applied Chemistry and Chemical Engineering, Rajshahi University, Rajshahi 6205, Bangladesh
5

6
20 a r t i c l e i n f o a b s t r a c t
7
8 Article history: Alpha-cellulose extracted from corn husks was used as the raw material for the production of food-grade
9 Received 9 February 2015 carboxymethyl cellulose (CMC). Preparation of CMC from husk cellulose was carried out by an etherifica-
10 Received in revised form 17 April 2015 tion process, using sodium hydroxide and monochloroacetic acid (MCA), with ethanol as the supporting
11 Accepted 23 April 2015
medium. Characterizations of CMC were carried out by analyzing the spectra of FTIR, XRD patterns and
12 Available online xxx
SEM photomicrographs. Degree of substitution (DS) was determined with respect to particle size using
13
chemical methods. Solubility, molecular weight and DS of CMC increased with decreased cellulose particle
14 Keywords:
sizes. Microbiological testing of the prepared CMC was done by the pour plate method. Concentrations
15 Cellulose
16 Carboxymethyl cellulose
of heavy metals such as arsenic, lead, cadmium and mercury in the purified CMC were measured by
17 Corn husk Atomic Absorption Spectroscopy technique and found to be within the WHO/FAO recommended value.
18 Food grade A comparative study with CMC available in the international market was conducted. The purity of the
19 Etherification prepared CMC was higher, at 99.99% well above the purity of 99.5% for standard CMC. High purity CMC
showed a yield 2.4 g/g with DS 2.41, water holding capacity 5.11 g/g, oil holding capacity 1.59 g/g. The
obtained product is well suited for pharmaceutical and food additives.
© 2015 Published by Elsevier B.V.

21Q3 1. Introduction prepared by the reaction of monochloroacetic acid with alkali cel- 39

lulose [4]. 40

22Q4 Cellulose is a common natural polymer. It can be found widely There are several grades of CMC depending on their applica- 41

23 in plants which are used as raw material for producing modified tions such as technical, semi-purified and purified. Purified CMC is 42

24 cellulose. Due to the abundant supply of the polymer in nature, a white to cream colored, tasteless, odorless, free-flowing powder 43

25 modified cellulose is now advancing in terms of production and [5] and used in a variety of industries including the food, deter- 44

26 innovation. The sources can range from wood even to the agricul- gents, personal care, pharmaceutical etc. [6,7]. High purity grades 45

27 tural waste. Corn or maize husk is an agricultural waste obtained are employed as food additives [4] also known as cellulose gum 46

28 from corn fields which is a rich source of cellulose. [8]. In the food industry, it can be used as suspending agent, water 47

29 Cellulose is a linear, high molecular weight, biodegradable loss reducer, thickener, emulsifying agent and stabilizer or dispers- 48

30 polymeric material. However, due to its strong inter- and intra- ing agent. It is used as a preservative for coating of fresh fruit and 49

31 molecular hydrogen bonds, cellulose neither melts nor dissolves thickener for pharmaceutical products [9–11]. 50

32 readily in hot or cold water [1] and in most common organic Some authors have reported the synthesis of CMC from various 51

33 solvents [2,3]. In order to utilize cellulose in the food indus- cellulosic sources such as raw cellulose [7], paper sludge [12], wood 52

34 try, cellulose must be converted into its derivatives. One of the residue [13], cotton linters [14,15], fibers [16] etc. There is consid- 53

35 most common derivatives is carboxymethyl cellulose (CMC), also erable interest in finding cheaper alternative methods to produce 54

36 referred to as Na-CMC which is currently finding an increasing CMC. Recently cultivation of corn has tremendously increased in 55

37 number of applications. CMC is manmade modified cellulose, a Bangladesh [17] and huge amounts of corn husks are either thrown 56

38 linear, long-chain, water-soluble, anionic polysaccharide which is away as waste or burnt. However, these are applications with low 57

added value, causing disposal as well as environment pollution 58

problems. 59

The purpose of the present research is to investigate the suitabil- 60

Q2 ∗ Corresponding author. Tel.: +880 1914254992; fax: +880 721750064.
ity of the corn husk as a source to produce high purity food-grade 61
E-mail addresses: mihmondal@yahoo.com, mihmondal@gmail.com
(Md.I.H. Mondal). CMC. Every year in Bangladesh, large amounts of CMC are being 62

http://dx.doi.org/10.1016/j.ijbiomac.2015.04.061
0141-8130/© 2015 Published by Elsevier B.V.

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63 imported to meet domestic demand and the importance of CMC is 2.3. Characterization 119

64 increasing day-by-day [18]. Therefore, there is a need to produce

65 CMC from locally available cheap raw material, such as corn husk 2.3.1. Measurement of CMC yield 120

66 agricultural waste, economically on a large scale. The novelty of this CMC yield was measured based on a dry weight basis. The net 121

67 study is the synthesis of CMC with higher DS as well as higher purity weight of dried CMC was divided by the weight of cellulose to get 122
123
68 so that it can be used for food-based products and pharmaceuticals. the yield value [21], as follow:

Weight of prepared CMC (g)

CMC yield (%) = × 100 124
69 2. Materials and methods Weight of dried cellulose (g)

70 2.1. Materials
2.3.2. Determination of degree of substitution 125

71 Corn husks were collected from the Wheat Research Insti- To determine the degree of substitution, 0.5 g of dried sodium 126

72 tute, Rajshahi, Bangladesh. Chemicals used during the study CMC was ashed gently between 450 and 550 ◦ C for 24 h, and then 127

73 were sodium hydroxide (Merck, India), monochloroacetic acid dissolved in 100 mL of distilled water. 20 mL of this solution was 128

74 (BDH, England), standard CMC (BDH, England), ethanol (Merck, titrated with 0.1 N sulphuric acid using methyl red as an indicator. 129

75 Germany), methanol (Merck, Germany), glacial acetic acid After the first end point, the solution was boiled and titrated to a 130

76 (BDH, England), silver nitrate (BDH, England), n-hexane (Merck, sharp end point. The carboxymethyl content was calculated from 131
132
77 Germany), sodium chlorite (Merck, Germany), etc. All chemicals the degree of substitution [22], as follow:
78 used were of reagent grade and used without further purification. 0.162 × B
Degree of Substitution (DS) =
1 − 0.08 × B
133
79 2.2. Methods 0.1 × b
B=
G
80 2.2.1. Preparation of sample
81 Locally collected corn husk samples were cut manually into where, b is the volume (in mL) of 0.1 N sulphuric acid and G is the 134

82 small pieces and dried in the sun in order to remove moisture. mass of pure CMC in grams. 135

83 The dried sample was ground into powder using a grinding disk
84 mill (model: FFC-15). The powdered sample was then sieved (sieve 2.3.3. Determination of molecular weight 136

85 type: AFNOR X11-501), and separated into different particle sizes, CMC was dissolved in a 0.78 M NaOH solution and the molecu- 137

86 by passing the sample through sieves of different mesh sizes (e.g. lar weight was determined using an Ostwald viscometer. From the 138

87 10, 44, 100, 150 and 200 mesh) and stored in a desiccator for com- intrinsic viscosity, the molecular weight of the CMC was calculated 139
140
88 positional and chemical analysis. by the “Mark–Houwink–Sakurada” equation [23], as follow:

[] = K M a 141
89 2.2.2. Isolation of ˛-cellulose
90 A suitable amount of powdered husk sample was treated with where K is the constant for solvent, “a” is the polymer shape factor, 142

91 18% NaOH solution for 2 h with occasional stirring in a solid to [] is the intrinsic viscosity and M is the molecular weight of CMC. 143

92 liquor ratio of 1:100 at room temperature. The cellulose residue

93 was separated by filtration, washed thoroughly with 2% acetic acid 2.3.4. Water retention capacity and oil retention capacity 144
94 and then with hot distilled water. The cellulose was stirred with 25 mL of distilled water or commercial olive oil were added to 145
95 n-hexane for 1 h at room temperature followed by washing with 1 g of dry sample, stirred and incubated at 40 ◦ C for 1 h. After cen- 146
96 95% ethanol. After filtration, the cellulose thus obtained was heated trifugation, the residue was weighed, and water retention capacity 147
97 with 0.7% NaClO2 solution buffered at pH 4 in the solid to liquor (WRC) and oil retention capacity (ORC) was calculated as gram 148
98 ratio of 1:50 at 90–95 ◦ C for 90 min. The ␣-cellulose was separated water or oil per gram of dry sample, respectively [24]. 149
99 by filtering and washing with ethanol and distilled water. It was
100 then treated with 0.2% sodium meta bisulphite solution for 15 min, 2.3.5. Moisture content and ash content 150
101 filtered, washed thoroughly with distilled water, and finally dried Moisture content in the CMC was measured by Moisture Ana- 151
102 at 60 ◦ C until reaching a constant weight [19]. lyzer (model: MAC 50/NH, Rad wag 26-600 Radom, Poland). Ash 152

content was determined using standard methods [25]. Approxi- 153

103 2.2.3. Synthesis of carboxymethyl cellulose mately 0.5 g of the vacuum dried sample was taken into the dried 154

104 Synthesis of CMC was carried out by two reactions: alkaliza- crucible and the sample was pre-ashed in a fume hood. When the 155

105 tion and etherification. The alkalization reaction was conducted sample ceased giving off smoke, it was placed in a preheated 600 ◦ C 156

106 at 30 ◦ C. First, a 30% (w/v) NaOH solution was infused into 5 g muffle furnace for 6 h. After completing the sample ashing, ash 157
158
107 of pure cellulose. A cellulose-to-liquor ratio of 1:2.7 was created content was calculated using the equation below:
108 with mechanical stirring for an hour. 150 mL ethanol was used as wt. of ash × 100
109 solvent in this step. Then, the etherification reaction commenced. Ash content (%) = 159
wt. of sample
110 120% (w/v) monochloroacetic acid (MCA) was added drop-by-drop,
111 under constant stiring, for 30 min to create a cellulose–liquor ratio
112 of 1:1.2. This step was continued for 3.5 h at 55 ◦ C. The product thus 2.4. Gel content 160
113 obtained was then filtered and suspended in 200 mL of methanol.
114 The slurry was neutralized using glacial acetic acid. Then the sam- Gel content of the CMC film was determined by Soxhlet extrac- 161
115 ple was washed, first in 70% ethanol solution four times and then tion using hot toluene as solvent. Films (6 cm × 4 cm) were weighed 162
116 with absolute ethanol, to remove undesired byproducts. Finally, the and placed in a cellulose extraction thimble in the Soxhlet extrac- 163
117 sample was filtered and dried in the oven (model: FC-610, Toyo, tor. The solvent extraction was carried out with 250 mL toluene for 164
118 Japan) at 60 ◦ C temperature [20]. 6 h. After that, the sample was vacuum dried and re-weighed until 165

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166 it attained a constant weight. Gel content was calculated according Table 1
167 Preparation of CMC from corn husk cellulose in aqueous ethanolic medium with dif-
to the following equation [26]:
ferent particle sizes (30% NaOH in the cellulose–liquor ratio, 1:2.7; 120% ClCH2 COOH
W1 in the cellulose–liquor ratio, 1:1.2; temperature, 55 ◦ C; time, 3.5 h).
168 Gel Content (%) = × 100
W0 Particle size Degree of Yield of CMC Solubility
(␮m) substitution (DS) (%)
169 where W0 is the weight before extraction and W1 is the weight after
1071 0.21 100.89 Insoluble in water
170 extraction.
340 0.38 115.75 Insoluble in water
149 1.52 195.51 Highly soluble in water
171 2.5. CMC content 100 2.21 210.87 Highly soluble in water
74 2.41 240.58 Highly soluble in water

172 Exactly 1.5 g of CMC was added to 100 mL of 80% aqueous

173 methanol solution. This mixture was stirred, kept for 10 min and count was used to detect all viable micro-organisms growing aer- 215
174 filtered. The cake was washed with 100 mL of fresh 80% aqueous obically, as reflecting the general hygiene condition of a sample. 216
175 methanol and dried to obtain pure CMC [10]. The CMC content was Nutrient agar media was used for total plate count. 217
176
calculated as follows:
W 2.9. Atomic absorption spectroscopy 218
177 CMC content (%) = × 100
W0
The amount of arsenic (As), lead (Pb), cadmium (Cd) and mer- 219
Q5
178 where W0 (g) is the weight of sample before washing and W (g) is cury (Hg) in the prepared CMC was measured by Atomic Absorption 220
179 the weight of washed sample. Spectroscopy (Model: AA-68000, Shimadzu, Japan) coupled with 221

an auto-sampler (ASC-6100, Japan) employing absorption of opti- 222

180 2.6. NaCl content in CMC cal radiation by free atoms in the gaseous state. Metal ions in the 223

sample was quantitatively determined by measuring the radiation 224

181 2 g of CMC was added to 250 mL of 65% aqueous methanol and absorbed by atoms of the sample solution with compared to the 225
182 kept for 5 h. 100 mL of this mixture in a liquid form was neutralized amount absorbed by a known concentration. 226
183 by diluted 0.1 N HNO3 and titrated with 0.1 N AgNO3 solution [10].
184
The NaCl content was calculated as follows: 2.10. FTIR spectroscopy 227

1.461 × V
185 NaCl (%) = Infrared spectra of the prepared CMC samples were recorded 228
M
with FTIR (Model: FTIR-8900, Shimadju, Japan). Pellets were 229
186 where V (mL) is the amount of AgNO3 solution and M (g) is the made from 0.2 mg CMC samples, which were ground with 2 mg 230
187 weight of the dried sample. KBr. Transmission was measured at the wavenumber range of 231

4000–400 cm−1 . 232

188 2.7. Sodium glycolate content

2.11. X-ray diffractometry 233
189 0.5 g of CMC was moistened with 5 mL glacial acetic acid and
190 5 mL water. 50 mL acetone and 1 g sodium chloride was added Diffraction diagrams of samples were recorded between 10◦ and 234
191 to the mixture and stirring was continued for several minutes to 50◦ using a Bruker D8 Advanced Germany X-Ray Diffractometer 235
192 ensure complete precipitation of CMC. The mixture was then fil- that generated CuK␣ radiation. Powder samples were exposed to 236
193 tered and clear supernatant was used to prepare the test solution. A X-ray beam (40 kV, 30 mA) at 2◦ /min. 237
194 similar blank solution, using only reagents, was prepared. Acetone
195 was removed from both the test solution and the blank solution 2.12. Scanning electron microscopy 238
196 by heating both for 20 min in a boiling water bath. The solutions
197 were cooled. Then 20 mL of 2,7-dihydroxynaphthalene was added A thin sample film was coated with gold using an ion sputter. 239
198 to each solution. Absorbance of the test solution, compared with The coated samples were viewed and photographed with a scan- 240
199 that of blank solution, was measured at 540 nm wavelength using ning electron microscope (Model-S 3400 N, VP SEM, Hitachi, Japan) 241
200 a UV–vis spectrophotometer (model: T 60, England). A calibration using 20 (kV) accelerating voltage. 242
201 curve of pure standard glycolic acid was used to determine the
202 unknown concentration of sodium glycolate in the solution using 3. Results and discussion 243
203
the following formula [27]:

a × 1.29 Carboxymethyl cellulose was prepared from corn husks cellu- 244
204 Sodium glycolate, % = lose according to Scheme 1. The advantage of this process was the 245
b
use of recovered ethanol through distillation which was above 90%. 246
205 where 1.29 is a factor for converting glycolic acid into sodium gly- The reaction was optimized with respect to DS by varying each 247
206 colate, a is mg of glycolic acid read from the calibration curve and of the parameters. The yield of CMC at different cellulose particle 248
207 b is the dry weight of sample in g. sizes is shown in Table 1. It can be observed from Table 1 that % 249

yield of CMC increased with decrease of particle size and that the 250

208 2.8. Microbiological test highest yield obtained was 240.58% with respect to particle size 251

74 ␮m. Decreased cellulose particle size provides a greater surface 252

209 The presence of microorganisms such as yeasts and molds, area which increases the chance of collisions between reactants and 253

210 Escherichia coli, Coliform and Salmonella in the prepared CMC was cellulose particles. Thus, the reaction rate increases as well as the 254

211 determined using the following method described in the literature. production of CMC [29]. The solubility of the prepared CMC sam- 255

212 Lactose broth was used for the detection of Coliform, Salmonella and ples with different particle sizes as well as different DS was tested. 256

213 E. coli bacteria as per Standard Methods [28]. Potato dextrose agar It was found that CMC, with DS 0.21 and 0.38, was insoluble in 257

214 media was used for yeast and mold detection. The test of total plate water but CMC, with DS 1.52 to 2.41, was highly soluble in water. 258

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Scheme 1. Flow chart for the preparation of CMC from corn husk cellulose.

259 CMC, with DS less than 0.40, is insoluble in water. Above this value the fact that a greater number of carboxymethyl groups were sub- 287

260 of DS, CMC is fully soluble, with its hydro affinity increasing with stituted for the hydroxyl groups of the cellulose molecules. These 288

261 increasing DS [30]. carboxymethyl groups are hydrophilic. Therefore, an increase of DS 289

262 From Table 2, it can be seen that the DS, intrinsic viscosity and increases the ability of CMC to immobilize water in a system [20]. 290

263 molecular weight of the prepared CMC increased gradually with The results of moisture content, ash content, gel content, WRC 291

264 the decrease of cellulose particle sizes. The highest DS obtained was and ORC of standard CMC and that of prepared CMC are listed in 292

265 2.41 with respect to particle size 74 ␮m. Smaller cellulose particle Table 3. From Table 3, it can be seen that moisture content and ORC 293

266 sizes have larger surface area, so excess reactants can penetrate of standard CMC and prepared CMC are almost the same. However, 294

267 into the cellulose at the same time. Etherification mainly depends ash content, gel content and WRC of prepared CMC are higher than 295

268 upon the accessibility of reactants and the availability of the acti- that of standard CMC. The high WRC implies that prepared CMC 296

269 vated hydroxyl groups [18]. When particle size decreases, surface is highly hydrophilic. During etherification, more hydroxyl groups 297

270 area as well as the number of available free OH groups for substi- were replaced by carboxymethyl groups, which are hydrophilic 298

271 tution reaction increases, thus DS increases. Reduction in particle [20]. The high gel content implies high reactivity [26]. The high ash 299

272 size of cellulose could enhance the affinity between cellulose par- content implies high DS, perhaps because more hydroxyl groups 300

273 ticles and reactants. Thus such smaller particle size increases the are substituted by sodium salts of carboxymethyl groups during 301

274 etherification rate as well as the carboxymethyl substitution rate the etherification reaction. 302

275 [31]. The molecular weight of the prepared CMC increased with
276 the increase of DS. Such molecular weight also increased with the 3.5
277 decrease of particle size. As the DS increased the number of OH
3
Intrinsic viscosity

278 groups were replaced by heavier carboxymethyl groups. As a result

279 the molecular weight of the final product CMC increased [18]. 2.5
280 Viscosity of a polymer solution depends mainly on the concen- 2
281 tration and size (i.e., molecular weight) of the dissolved polymer. 1.5
282 Thus more the intrinsic viscosity, larger the polymer size as well as 1
283 the higher the molecular weight. 0.5
284 There is a direct relationship between intrinsic viscosity and DS
0
285 of the prepared CMC. Intrinsic viscosity increases with the increase
0.21 0.38 1.52 2.21 2.41
286 of DS as presented in Fig. 1. The effect of DS on viscosity was due to
Degree of Substitution

Fig. 1. Relationship between degree of substitution and intrinsic viscosity of pre-

Table 2
pared CMC.
Determination of DS and molecular weight of prepared CMC as a function of particle
sizes (molecular weight was calculated from equation [] = KMa where, value of
k = 37 × 10−5 dL/g and a = 0.61 at 35 ◦ C). Table 3
Determination of water holding capacity (WHC), oil holding capacity (OHC), mois-
Particle size Degree of Intrinsic Molecular ture, ash and gel contents of synthesized CMC (DS = 2.41) and standard CMC
(␮m) substitution (DS) viscosity () weight (Da) (DS = 0.8).
1071 0.21 0.36 79,163
Samples name Moisture Ash WHC OHC Gel content
340 0.38 0.54 153,886
content (%) content (%) (g/g) (g/g) (%)
149 1.52 2.21 1,550,516
100 2.21 2.64 2,075,165 Standard CMC 2.22 14.23 4.12 1.58 99.06
74 2.41 3.00 2,558,967 Prepared CMC 2.21 18.05 5.11 1.59 99.96

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Table 4 Table 6
Physicochemical characteristics of corn husk CMC. CMC grades and typical applications (6).

Tests CMC Quality of CMC Examples of Content of Content of

application areas CMC, % salts, %
Organoleptic Odorless, white and tasteless with free flowing
powder Technical Detergents, mining <75 >25
Solubility Forms highly viscous solution with water but flotation
insoluble in ethanol Semi-purified Oil and gas drilling 75–85 15–25
Foam No layer of foam appears by shaking 0.1% muds
solution Purified Paper coating, >98 <2
pH in 1% solution 6.8 textile sizing and
Starch and dextrin’s No blue or reddish brown color with iodine printing, ceramic
solution glazing, oil drilling
Organic impurities No red color with acidified phlorogucinol muds
Film formability Able to form film Extra purified Food, toothpaste, >99.5 <0.5
(cellulose gum) pharmaceuticals

303 The physicochemical test results of prepared CMC obtained from

The culture responses of tested organisms in the prepared CMC 343
304 corn husk are listed in Table 4. Tested results in Table 4 show that
are given in Table 8. The total plate count is intended to indicate the 344
305 produced CMC is white in color, fine powdered, odorless, taste-
number of microorganisms in a product. No colonies were observed 345
306 less, freely soluble in water and insoluble in ethanol. The pH of 1%
in a sample containing cultured media after 48 ± 2 h incubation at 346
307 CMC solution was 6.80. No layer of foam appeared after shaking a
35 ± 1 ◦ C. Where no colonies are visible, the result is expressed as 347
308 0.1% solution of the CMC sample. This test distinguishes CMC from
less than 100 cfu/g [28]. 348
309 other cellulose ethers and from alginates & natural gums [32]. No
Yeasts grow as creamy-to-white colonies and molds grow as fil- 349
310 blue or reddish brown color of CMC developed in an iodine solution
amentous colonies of various colors on Potato Dextrose Agar (PDA) 350
311 or red color with acidified phlorogucinol. These tests confirm the
media. When a 1 g sample aliquot was poured on to a plate con- 351
312 absence of starch, dextrin and organic impurities in the prepared
taining PDA, no colonies were visible after 5 days incubation at 352
313 CMC. The Joint FAO/WHO Expert Committee on Food Additives [33]
20–25 ◦ C. This confirms the absence of yeasts and molds in the 353
314 has established some standard for the identification and purity of
sample. 354
315 CMC. The prepared CMC in this study meets these requirements.
E. coli grows as dark blue to violet colonies, Coliform as salmon to 355
316 Heavy metals are potential environmental contaminants, with
red colonies and Salmonella as a colorless colony. Results in Table 8 356
317 the capability to cause human health problems, due to their toxic
also shows that Coliform, Salmonella and E. coli either failed to grow 357
318 effects, even at very low concentrations [34]. Toxic metal content
or produced negative results, after 24 ± 2 h incubation at 35 ± 1 ◦ C, 358
319 in the prepared CMC was determined and compared with the rec-
in a plate containing a 1 g sample in 99 mL lactose broth. 359
320 ommended values shown in Table 5. From the results in Table 5,
321 it can be observed that the concentrations of Pb, Cd, Hg and As
3.1. FTIR analysis 360
322 are 0.0001, 0.0005, 0.001 and 0.0002 ppm, respectively. These val-
323 ues are within the World Health Organization’s (WHO) permissible
Characterization of extracted cellulose was done by FTIR spec- 361
324 limits for food additives and emulsifier.
troscopy. The differences in result between the raw corn husk 362
325 The purity of the CMC was measured after washing the prod-
powder and the extracted cellulose powder are shown in the 363
326 uct several times with 70% ethanol and absolute ethanol. Washing
FTIR spectra presented in Fig. 2. The main differences were found 364
327 removed the reaction by-products such as sodium chloride and
in the region from 1800 to 1000 cm−1 . The absorption bands at 365
328 sodium glycolate. Where the CMC product is intended for human
1740.13, 1638.49, 1605.18 and 1514.15 cm−1 were not observed 366
329 consumption, CMC must be purified to a level of 99.5% [35]. Table 6
in the extracted cellulose. Particularly two absorption bands must 367
330 shows the different grade of CMC with their applications and also
be emphasized: the bands at 1514 and 1249 cm−1 . The band 368
331 shows that only extra-purified CMC is used as food additive. The
at 1514 cm−1 was not present and the band at 1249 cm−1 was 369
332 percentages of CMC content and sodium salts in the prepared CMC
drastically reduced on the purified cellulose spectra. These two 370
333 were determined and compared to that of standard CMC and these
absorption bands were important because their absence in the 371
334 result are summarized in Table 7. It can be observed from Table 7
extracted cellulose spectra strongly indicates that most of the lignin Q6 372
335 that the percentages of sodium chloride and sodium glycolate in
had been removed [38,39] (Fig. 3). 373
336 the prepared CMC are very low, 0.009% and 0.001%, respectively.
337 Sodium glycolate is toxic in nature. It is reported in the literature
338 [27,33] that sodium chloride and sodium glycolate content in food Table 7
339 additives should not exceed 0.5% individually or in combination. Determination of CMC, NaCl and sodium glycolate contents.

340 The purity of prepared CMC was higher, at 99.99%, well above the Parameters Content of CMC Content of Content of sodium
341 purity of 99.5% required for standard CMC and 98% required for or purity, % NaCl, % glycolate (%)
342 commercial grade CMC. CMC prepared 99.99 0.009 0.001
CMC standard 99.95 0.03 0.02

Table 5
Concentration of heavy metals in the synthesized CMC. Table 8
Microbiological test of the prepared CMC.
Heavy metal Concentration in Concentration in ppm
ppm (studied (proposed permissible Tested organisms Cultural response
values) limit) [36,37]
Total plate count (cfu/g) 100
As 0.0001 3 Mold (cfu/g) 100
Pb 0.0005 1 or lower in case of Yeast (cfu/g) 100
high consumption Colliform (g) Negative
Cd 0.001 1 Salmonella (g) Negative
Hg 0.0002 1 E. coli (g) Negative

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Fig. 2. FTIR spectra of (a) raw corn husk powder and (b) purified extracted cellulose.

374 The infrared spectra of prepared CMC and standard CMC sam-
375 ples have been shown in Fig. 2. It is evident from Fig. 2b that the
376 broad absorption band at 3434.92 cm−1 is due to the stretching fre-
377 quency of the OH group and a band at 2924.07 cm−1 is attributable
378 to C H stretching vibration. The presence of a new and strong
379 absorption band at 1620.16 cm−1 confirms the stretching vibration
380 of carboxyl groups (COO− ) and 1423.81 cm−1 is assigned to car-
381 boxyl groups as salts [40,41]. The bands around 1329.32 cm−1 and
382 1112.65 cm−1 are assigned to -OH bending vibration and C O C
383 stretching, respectively. Wavelength 894 cm−1 detected 1,4-␤ gly-
384 coside of cellulose [42]. The IR spectra of the standard CMC procured
385 from the market are also recorded in Fig. 2a and the patterns of two
386 spectra, prepared CMC and standard CMC are almost similar.
Fig. 4. X-ray diffractogram of (a) pure husk cellulose and (b) carboxymethylated
cellulose (DS = 2.41).

387 3.2. XRD analysis

the small crystallites in cellulose granules, which conforms the 394

388 X-ray diffraction (XRD) analysis is a special technique for theory of XRD. This theory states that very small imperfect crystals 395

389 estimating the degree of crystallinity in polymer. Cellulose is give broadened diffractions. 396

390 semi-crystalline in nature. Fig. 4 shows the diffraction pattern The diffraction patterns of CMC show a destruction of the crys- 397

391 of pure husk cellulose and carboxymethylated cellulose. The talline structure of the original cellulose (Fig. 4). All characteristic 398

392 peaks correspond to the crystalline phase and the background peaks for native cellulose have almost disappeared and trans- 399

393 corresponds to the amorphous phase. The peaks are broad due to formed into an amorphous phase. Therefore CMC has excellent 400

Fig. 3. FTIR spectra of (a) standard CMC (DS = 0.8 and (b) synthesized CMC (DS = 2.41).

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Md.I.H. Mondal et al. / International Journal of Biological Macromolecules xxx (2015) xxx–xxx 7

Fig. 5. SEM image of (a) cellulose and (b) CMC at 500 magnification (200 ␮m size bar).

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402 ity [12]. During the carboxymethylation process, the cellulose [14] Z. Nagieb, M.E. Sakhawy, K. Samir, Int. J. Polym. Mater. 50 (2001) 163–173. 445
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