Pergamon

Materials Research Bulletin 36 (2001) 511519

Biopolymer composite lms based on -carrageenan and chitosan

Sun Y. Parka, Burtrand I. Leeb,*, Soon T. Jungc, Hyun J. Parkd
b

Currently at Department of Packaging Science, Clemson University, Clemson, SC 29634-0370, USA Department of Material Science & Engineering, Olin Hall, Clemson University, Clemson, SC 29634-0907, USA c Department of Food Engineering, Mokpo National University, Muan 534-729, Korea d Graduate School of Biotechnology, Korea University, Seoul 136-701, Korea and the Department of Packaging Science, Clemson University, Clemson, SC 29634-0370, USA (Refereed) Received 23 May 2000; accepted 18 September 2000

Abstract Bioploymer composite lms of -carrageenan and chitosan were prepared by co-dissolving -carrageenan and chitosan in several different organic acids. Tensile strength (TS), elongation (E), and water vapor permeability (WVP) were determined as a function of ascorbic acid added. Ascorbic acid tends to increase the properties of the lms from all acids. Malic acid exhibited largest increase in tensile strength and elongation at 2% ascorbic acid and lactic acid in water vapor permeability. The strongest acid, citric acid exhibits overall the worst properties regardless of the amount of ascorbic acid. The ascorbic acid generally showed increase in TS and %E but little effect on the WVP except that from acetic acid. The results of the properties of the composite lms indicated that there are interactions among the organic solvents, the plasticizer, -carrageenan, and chitosan in a complex manner. 2001 Elsevier Science Ltd. All rights reserved.
Keywords: A. Polymers; B. Sol-gel chemistry; D. Mechanical properties

1. Introduction Over the past several decades, several biopolymers have received increased attention for their applications in chemical, biomedical, and food industries [13]. For example, chitin
* Corresponding author. E-mail address: lburtra@clemson.edu (B.I. Lee).
0025-5408/01/$ see front matter 2001 Elsevier Science Ltd. All rights reserved. PII: S 0 0 2 5 - 5 4 0 8 ( 0 1 ) 0 0 5 4 5 - 1

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suture is resorbable in human tissues from which chitosan-collagen composites for an articial skin are produced commercially. These polymers are not only biodegradable, but also edible. Hosokawa et al. [4] have reported that the chitosan and cellulose composite lms have a low oxygen permeability (2 8 ml/m2dayatm), which is much lower than that of polyethylene lms but is compatible with that of nylon or polyester lms. Butler et al. [5] reported that chitosan-based lm has high mechanical strength (1530 M2Pa) and is an excellent oxygen barrier property (4.6 105 cm3/mdayatm). Bai et al. [6] have found that chitosan-acetic acid complex membrane showed high selectivity for oxygen and carbon dioxide permeation. The separation factors (CO2/O2) of these membranes were much smaller than unity, indicating possible applications for the preservations of fruits and vegetables. Chitosan is a polysaccharide obtained from the deacetylation of chitin, the second-most abundant biopolymer present in nature after cellulose. Chitin has repeating structure of poly--(134)-N-acetyl-D-glucosamine and is especially abundant in marine invertebrates, insects, fungi and yeasts. Chitosan is insoluble in water, but becomes soluble and cationic when it is dissolved in acidied solutions using acetic acid, citric acid, formic acid, or hydrochloric acid, while most other biopolymers exhibit anionic property in solvents. Since most other soluble biopolymers become anionic in water, chitosan cations exhibit good afnity for other biopolymers. From the excellent lm forming properties of chitosan and cellulose a novel degradable plastic from chitosan and cellulose has been developed in Japan [3,4]. Tensile strength (TS) of the composite plastic was 1000 kg/cm2, 5 times higher than that of polyethylene. Carrageenans are water soluble polymers extracted from red algae. They are used in food and pharmaceutical industries as gelling and stabilizing agents, microencapsulation and immobilization of drugs and enzymes. Carrageenans consist of alternating copolymers of -(133)-D-galactose and -(134)-3,6-anhydro-D- or L-galactose. Several isomers of carrageenan are known as -, -, and -carrageenan, and they differ in the number and position of the ester sulfate groups on the repeating galactose units [7]. -carrageenan has only one negative charge per disaccaharide with a tendency of forming a strong and rigid gel. -carrageenan has two and -carrageenan bears on the average 2.7 charges per disaccharide unit. The gelling power of -and -carrageenans impart excellent lm forming properties. From these properties composite lms utilizing polyelectrolyte behavior of -carrageenan and chitosan with opposite ionic charges were prepared in our laboratory. The mechanical and barrier properties of the oppositely charged composite biopolymer lms have further been improved by cross-linking among the biopolymer molecules by using enzymes [8 10], chemical treatment [1113], or heat curing method [14 16]. Lee et al. [17] have shown a strengthening effect in corn protein lms by incorporating polymeric silica gel on the expense of elongation properties. Ascorbic acid is known to have a protective effect against the browning developed in bioprocess because of its reducing power, and it could be used as a cross-linking agent between molecules of carrageenan and chitosan [18]. Our previous results on the effective afnity of ascorbic acid on the chitosan and -carrageenan separately are given in Table 2 [21]. TS of the chitosan-based lms cast from acetic acid was 3.2 4.7 MPa, increased with increasing concentration of ascorbic acid. E and WVP each 25.5, and 16.7% decreased increasing with concentration of ascorbic acid

S.Y. Park et al. / Materials Research Bulletin 36 (2001) 511519 Table 1 Physical constants and molecular formulae of acids used Acid pKa MW (g/mol) Molecular formula

513

MP (C)

Table 2 Mechanical properties and water vapor permeability of chitosan and -carrageenan lms as a function of ascorbic acid Films Chitosan 0 1 2 3 0 1 2 3
a,b,c

Ascorbic acid conc. (%)

TS (MPa)

E (%)

WVP (ng.m/m2sPa) 3.6a 0.2 3.4a 0.1 3.1a 0.1 3.0a 0.2 1.9a 0.03 1.8a 0.03 1.8a,b 0.02 1.6b 0.02

-Carrageenan

3.2c 0.7 3.8b 0.6 4.6a 0.9 4.7a 0.8 30.2b 13.1 34.2a,b 10.1 37.8a 9.8 37.8a 8.4

84.2a 9.7 75.3b 12.6 66.3c 9.1 62.7c 9.5 5.0b 3.2 4.6b 3.2 7.7a 4.0 5.6a,b 0.6

The values in same column with different superscripts are signicantly different at p 0.05 by Duncans multiple range test.

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increased. TS of -carrageenan lms was 30 38 MPa and had high mechanical and barrier properties as they are compared with TS of chitosan (3.2 4.7 MPa). Especially, E of -carrageenan lms increased as the concentration of ascorbic acid increased. WVP of carrageenan-based lms was 1.6 1.9 ngm/m2sPa, comparable with those of other biopolymer and plastic lms. However, WVP of both the chitosan and -carrageenan lms decreased, while increase in TS, with increasing ascorbic acid. Thus the objectives of this study were; (a) to prepare composite lms based on chitosan and -carrageenan to gain some of the high mechanical strength of carrageenan and some of the superior biological properties of chitosan, (b) to measure the water vapor barrier properties and mechanical properties (tensile strength and elongation) of the lms, and (c) to evaluate the effectiveness of ascorbic acid as a crossing-linking and a reducing agent on mechanical and barrier properties of the lms.

2. Experimental The materials used to prepare carrageenan/chitosan composite lms were chitosan (red crab with degree of deacetylation, 95%; viscosity, 10 cP; from Biotech Company, Inc., Mokpo, Korea); -carrageenan (MSC Company, Yangsan, Kyungnam, Korea); polyethylene glycol, 400 (Junsei Chemical Co. Ltd., Tokyo, Japan); glycerin (USP grade Aldrich Chemical Company, Inc., Milwaukee, WI, USA); and organic acids such as acetic, lactic, citric, malic and ascorbic (Aldrich Chemical Company, Inc., Milwaukee, WI, USA). The molecular structure, pKa, molecular weight (MW), and melting pont (MP) of these acids are given in Table 1 [19,20]. The experimental ow diagram for fabrication of -carrageenan/chitosan composite lms is given in Fig. 1. -carrageenan (2% w/w) solutions were prepared by dissolving the carrageenan in distilled water and chitosan solutions by dissolving chitosan in a 2% acid solution. The composite lms were prepared in -carrageenan : chitosan 90:10 proportion with 50:50 (w/w) mixture of glycerin (GLY) and polyethylene glycol (PEG) as a plasticizer (0.75 g/sample g). Different amounts of the antioxidant acsorbic acid were added with continuous stirring. The composite lm solutions were poured onto a glass plate followed by drying for 12 hours at 70C in a drying oven. The dried composite lms were peeled from the plate and samples were cut into sizes for TS, E, and WVP. A hand-held micrometer (B. C. Ames Co., Waltham, MA) was used to measure lm thickness to an accuracy of 0.000127 cm. Five measurements were made on each test sample, and a mean thickness was calculated. Twenty specimen samples in 10 cm 2.54 cm cut from the cast lms on glass plates (28cm 28cm) were conditioned for 48 hrs at 25C and 50% relative humidity (RH) in an environmental chamber (Model 317332, Hotpack, Corp., Philadelphia, PA) before measuring TS and E. TS and E of samples were measured as quickly as possible. An Instron (Model 4210, Instron Engineering Corp., Canton, MA) was used to measure TS and E at break, according to the ASTM standard method D 882 88 [22]. Initial grip separation and crosshead speed were set at 50mm and 500mm/min, respectively. WVP of the lms was measured by the cup method, ASTM standard method E96-80 [23]. Cups made from polymethylmethacrylate (Piedmont Plastics, Inc., Greenville, SC) were

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Fig. 1. Flow diagrams for preparation of composite lms of -carrageenan and chitosan.

used to determine WVP. Cups consisted of cylindrical bottoms (ID 4.6cm, OD 8.7cm and depth 2.1cm) and sealing rings (ID 4.6cm, OD 8.7cm and thickness 0.5cm). Cups were lled to a depth of 0.9cm with distilled water and covered with a lm to be tested. Cups and sealing rings were tightened with four screws and placed in a chamber conditioned at 25C, 50% RH (Model 317332, Hotpack, Corp., Philadelphia, PA). An air ow meter (DatatrakTM, Model 635, Sierra Instrument Co., Carmel Valley, CA) was used to measure air ow rate inside the chamber. The air ow rate determined was 54.2ft/min. Weight change of the cups vs. time was measured and plotted. Linear regression was used to calculate the slope of a tted straight line. The WVP value corrected for the resistance of the stagnant air gap between underside of lm mounted on cups and surface of water inside the cups [24,25]. Duncans multiple range test was applied to compare the means for mechanical and barrier properties with a level of signicance of 0.05. The error bars in the gures represent the standard deviations for the plotted means.

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Fig. 2. Tensile strength of chitosan and -carrageenan composite lms prepared from different acid solvents as a function of ascorbic acid content. Acetic Acid, Citric Acid, Lactic Acid, } Malic Acid.

3. Results and discussion It is shown in Fig. 2 that all the organic acid solvents exhibited increasing tendency of TS as ascorbic acid additive increased. The trend was particularly remarkable for malic acid whose acidity expressed in pKa value is high among the acid solvents. However, citric acid whose acidity is the highest showed the least effect by adding ascorbic acid. The reactivity of ascorbic acid as a cross-linking agent for chitosan/carrageenan composite lms became ineffective. As the chitosan/carrageenan is bonded by electrostatic attraction between the negatively charged carrageenan and the positively charged chitosan, a high proton concentration from the acid solvent would protonate the sulfonate anionic groups of carrageenan. Then this will weaken the electrostatic bonding of the composite. This is shown by strongest acid citric acid exhibiting the lowest TS and the least cross-linking effect by ascorbic acid. The weakest acid acetic acid yielded the same TS as for malic acid within the experimental error in the absence of ascorbic acid but malic acid surpassed the TS of acetic acid at ascorbic acid above 1%. In Fig. 3 the elongation is also affected the most by the ascorbic acid addition in the composite lms made from the malic acid. The least effect by citric acid on E agrees with the least effect shown for TS by acid in Fig. 2. However, this is contrary to the usual behavior that TS and E are inversely related. This is also quite different from the trend of the data shown by Lee et al. [17] and Park et al. [26] in their corn zein and cellulose lms. Malic acid with high acidity not only increased the TS by aiding the role of ascorbic acid but also preserved the role of the plasticizer (GLY/PEG). Citric acid adversely affected the role of ascorbic acid as well as that of the plasticizer exhibiting both low TS and E. This indicate

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Fig. 3. %elongation of chitosan and -carrageenan composite lms prepared from different acid solvents as a function of ascorbic acid content. Acetic Acid, Citric Acid, Lactic Acid, Malic Acid.

the action of acid solvents and ascorbic acid are different for the carrageenan and chitosan as well as for the plasticizer. WVP of the biopolymer-based lms are generally 102103 higher than those of the typical plastic lms. In our previous study [27], various concentration and kinds of plasticizers greatly affect the WVP of biopolymer-based lms, i.e., WVP of both wheat gluten and corn-zein lms decreased as the increase in plasticizer. Fig. 4 shows the effect of different acid solvents on WVP of carrageenan/chitosan composite lms. However, the different acids showed little difference in WVP as ascorbic acid addition increased. The lowest WVP for malic acid, before ascorbic acid addition, agrees well with the highest TS in Fig. 2. WVP decreased by addition of ascorbic acid to chitosan and carrageenan lms (Table 2), yet ascorbic acid did not affect much the WVP in the composite lms. Park et al. [27] measured the MW of chitosan dissolved in different organic acid solutions by the light scattering method using an Otsuka light scattering instrument (DLS-7000). They found that the MW and molecular dimension of chitosan were different in the various acidic solutions. Therefore, matrix formation of the lm depends on the solvent system. It may be noted that the lm formed from acetic acid solution has tighter structure than those of the lms formed from the other acids. Gontard et al. [28] showed WVP increase as the plasticizer increase in gluten lms. At a constant plasticizer concentration for carrageenan/chitosan composite lms in this study, the lowest WVP was shown for malic acid, but as ascorbic increased, the WVP of the lm from malic acid increased about 40%. This agrees well with the explanation for E that malic acid preserved the role of the plasticizer, i.e., the more plasticizer, the higher the WVP. Lactic

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Fig. 4. The water vapor permeability of chitosan and -carrageenan composite lms prepared from different acid solvents as a function of ascorbic acid content. Acetic Acid, } Citric Acid, Lactic Acid, Malic Acid.

acid show the highest WVP with linear increase in of WVP as ascorbic acid increase. This, too, agrees well with E data in Fig. 3 with relatively low TS. Citric acid that exhibits the lowest TS and the lowest E, exhibited the intermediate WVP. The largest effect of lowering WVP, as the amount of ascorbic acid increased, came from acetic acid. The acetic acid as being the weakest acid with the smallest in molecular size and the lowest melting point, i.e., the most volatile, did not affect the role of ascorbic acid in all, TS, E, and WVP. Rather the WVP decreased generally by the ascorbic acid when the corresponding TS and E increased. This must mean that the ascorbic acid was not interfered by the solvent molecules. However, the increase in TS accompanied by increase E and decrease in WVP suggest that the ascorbic acid in acetic acid behaves the role of plasticizer in different way from the GLY/PEG as well as the role of the cross-linking agent.

4. Conclusions Controlling mechanical and barrier properties is essential to biotechnological use of biopolymer-based lms and plastics. Factors such as concentration, solvent system, and cross-linking agent can affect the biopolymer structure and, in turn, affect the mechanical and barrier properties. The mechanical strength, E, and WVP of -carrageenan/chitosan composite lms exhibited dependency on the organic acid solvent and ascorbic acid additive. A high acidity with high melting point malic acid exhibited superior overall properties: the highest TS and E, and low WVP. On the other hand, the strongest acid, citric acid, exhibited

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overall the worst properties regardless of the amount of ascorbic acid. The ascorbic acid generally caused increase in tensile strength and elongation of the lms, but little effect on the water vapor permeability except that in the presence of acetic acid.

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