European Polymer Journal 38 (2002) 13771381 www.elsevier.

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Xanthan gumgelatin complexes

C.-y. Lii
a b c

a,*

, S.C. Liaw a, V.M.-F. Lai b, P. Tomasik

a,c

Institute of Chemistry, Academia Sinica, Nankang, 11529 Taipei, Taiwan, ROC Department of Food and Nutrition, Providence University, Shalu, Taichung, Taiwan, ROC Department of Chemistry, University of Agriculture, Mickiewicz Avenue, 21, 31120 Cracow, Poland Received 30 May 2001; received in revised form 25 October 2001; accepted 15 November 2001

Abstract Gelatin (G) and xanthan gum (X) formed complexes either on bringing their blends to pH 2.3 or carrying out the electrode process in aqueous blends of X and G at pHs from 9 to 11. X carboxylic groups and G peptide moieties were involved in the interactions between the partners. Also non-coulombic interactions with involvement of NH and OH groups, as well as hydrophobic interactions were involved, as proved by FTIR and thermal analyses, respectively. The reaction yield declined with increase of concentration of G in the blends. Simultaneously, the thermal stability of the complexes slightly increased with increase in G content in the blend. The latter increased with its concentration in the reaction mixture. Slow formation by electrosynthesis provided more an organised matrix. 2002 Elsevier Science Ltd. All rights reserved.
Keywords: Coacervation; Electrosynthesis; Polysaccharideprotein complexes; Plant gums

1. Introduction For over two decades polysaccharideprotein complexes have received considerable attention for both theoretical and practical reasons [15]. Gelatin (G) is often used as protein component for complexes. It can complex to carboxymethyl cellulose (CMC) with involvement of electrostatic interactions [6]. Noncoulombic interactions were essential in the formation of complexes of G with pectins and alginates [7]. The character of the interactions was dependent on the reaction conditions, mainly on pH and ionic strength of the reaction mixture, as proved in complexes of G with CMC [8] and with gum Arabic [9]. Complexation of G with CMC was utilised in purication of these molecules [6] and complex formation with acacia gum was patented as a fat replacer in foodstus [10]. Complexes of G with acacia gum [1116], pectin [17], gellan gum [18], alginate [19], and CMC [19]

Corresponding author. Tel./fax: +886-2-2783-1237. E-mail address: cylii@chem.sinica.edu.tw (C.-y. Lii).

were considered as prospective sources for microcapsules. Xanthan gum (X) as a polysaccharide component of complexes with proteins was also subjected to numerous studies. In a complex with milk protein prepared in the pH range between 3.7 and 6.3, the components bound via hydrophobic interactions [20]. Complexes of X with milk proteins appeared to be interesting as potential shortening agents for bakery, a fat replacer [21], and an emulsier for food industry [22,23]. Complexes of X with soy protein [24,25], casein [24], ovalbumin [20,24], and whey protein [26] may be used as meat and fat replacers. Purication of whey proteins by complexation to X could be achieved [27]. In this paper, the synthesis and properties of the complexes formed from G and X are described. Simultaneously, the electrode process formerly applied for the synthesis of polysaccharideprotein complexes [2832] with covalent bonds between the partners is shown to be useful for slow coacervation, providing a more organised matrix of the resulting complexes. Although research on proteinpolysaccharide complexes has been addressed mainly in food science and

0014-3057/02/$ - see front matter 2002 Elsevier Science Ltd. All rights reserved. PII: S 0 0 1 4 - 3 0 5 7 ( 0 2 ) 0 0 0 0 8 - 3

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technology, the results of such studies might be potentially utilised for non-nutritional purposes. Such materials could serve as sizes for textiles, llers and thickeners for pulp and paper, coatings, microcapsules, and biodegradable constructing materials.

2. Results and discussion Electrosynthesis provided materials, which were 15 7% richer in G than the blends from which they were prepared. However, blends richer in G gave products with a lower excess of G. Generally, the yield of the products decreased with an increase in pH. High ratios of G in initial blends resulted in a decreased yield. The highest yield of the products separating on the anode was achieved from X:G 1:2 blends (Table 1). The current applied (Table 1) was automatically increased with pH. In the blends of 1:1 and 1:2 X:G ratios, the current rose with time. This might be an eect of a decrease in solution viscosity caused by the separation of the product. In the blends containing high ratio G, the current either remained unchanged, or decreased with time. The latter could result from the clathration of ions and/or water in these G-rich products.
Table 1 Composition and yields of XG complexes X:G ratio 1:1 Intended G, % 50 pH 9 10 11 9 10 11 9 10 11 9 10 11 Formed

The solubility tests for X, G, and the products prepared either by simple coacervation or electrosynthesis (Table 2), particularly solubility in 7 M aq. urea, revealed that coulombic interactions and formation of hydrogen bonds between X and G were involved. Thus, electrosynthesis did not promote formation of covalent bonds between those partners and the process taking place at the anode and in its vicinity was also a coacervation process. Coacervation of X with G took place at a pH of 2.3. Dierential FTIR spectra of 1:1 XG complex (Figs. 1 and 2) and with the subtracted spectrum of X taken at pH 2.3 (Fig. 2) showed that although the spectral patterns resembled a simple combination of the spectra of G and X, the essential band shifts could be observed. Contrary to X CMC formed complexes with G in which both partners were covalently bound [33]. X is a polysaccharide, the anionic property of which results from the presence of uronic acid moieties. The carboxylic groups in X are directly bound to the pyranose ring, and anionic properties of CMC result from the presence of the carboxylic groups bound to the glucose units through the 6CH2 OCH2 linkage. The dierences in the bonding of the carboxylic groups in X and CMC should not be reected by any essential dierences in their ionisation. Therefore, availability of the carboxylic

Yielda , % G ,% 58.2 57.8 57.5 72.6 75.5 73.9 82.1 80.1 75.9 87.2 84.3 85.0 75 73 68 77 76 73 67 65 55 62 58 54
b

Current change, A 0:01 ! 0:03 0:01 ! 0:03 0:04 ! 0:05 0:01 ! 0:02 0:01 ! 0:02 0:04 ! 0:03 0:01 ! 0:01 0:02 ! 0:02 0:05 ! 0:03 0:01 ! 0:01 0:02 ! 0:01 0:05 ! 0:02

Nitrogen content, % 10.1 10.0 9.9 12.6 13.1 12.8 14.2 13.9 13.1 15.1 14.6 14.7

1:2

67

1:3

75

1:4

80

a b

The yields were calculated on the basis of the total parent polymer weight with standard deviations of 0.33.2%. The data were calculated from the nitrogen content in complexes and original G sample (17.29%).

Table 2 Solubility tests of G, X, and XG complexes Sample GX complexes G X

a
a

Solvent H2 O 5% Na2 CO3 1% NaOH 7 M Urea DMSO 5% HCl

All complexes regardless the preparation method, initial composition of the blend and its pH.

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Table 3 FTIR spectral patterns of G (at pH 2.3), X (at pH 2.3) and 1:1 GX complex and the band assignments m cm1 1:1 GX complex 3416 s 3368 s 3081 2928 1719 1652 sh w m s 3081 w 2947 w 1652 s 1447 w 1404 w 1380 m 1333 w 1242 m 1152 w 1066 s 1033 s 1333 w 1252 m 1238 w 1152 m 1080 w 1061 s 1028 s G X 3416 s mOH intramolecular hydrogen bond mNH intramolecular hydrogen bond mOH polymeric association mCH mCOOH mC@O , dOH mCOO dCH dCH mCOO dNH dCH dNH ,mCN mC@O glucose units dCOH mCO , mCC , dCOH , C1H mCO , mCC , dOH , C4O Band assignment

2918 1733 1642 1538

w m m w

Fig. 1. FTIR dierential spectrum of the 1:1 GX product from simple coacervation (below). The spectrum of X taken at pH 2.3 was subtracted from the spectrum of coacervate and spectrum of G taken at pH 2.3 was added for comparison (above).

1447 w 1404 w

Fig. 2. FTIR dierential spectrum of the 1:1 GX product from simple coacervation (below). The spectrum of G taken at pH 2.3 was subtracted from the spectrum of complex and spectrum of X taken at pH 2.3 was added for comparison (above).

group for the complexing partner, which in X was much more obstructed by steric hindrances seemed to be crucial for the mode in which partners interact with one another. Electrosynthesis of GX complexes proceeded in the reaction mixtures of pH 9, 10 or 11 but the local pH around anode could be dierent. Indeed, the FTIR spectra of the 1:1 GX complexes from both processes were identical. Table 3 lists the wavelengths of the spectral bands and their assignments. The band shifts conrmed the involvement of the carboxylic groups of X and peptide bond moieties of G in the GX interactions. Simultaneously, the shifts of the vibration bands of the OH and NH groups showed that

non-electrostatic interactions might participate in coacervate formation. The pH at which the syntheses were carried out had no eect on the FTIR spectra, and varying proportions of X and G in the initial blends were reected exclusively by relative peak intensities. X was less thermally stable than G as shown by thermogravimetric (TG) and dierential thermogravimetric (DTG) analyses (Table 4). Thermal stability of the products from electrosynthesis slightly increased with an increase of G in product. The eect of pH upon the thermal stability of resulting products could be interpreted in terms of the eect of pH upon the content of G in the product. At higher pH, G content in the products decreased (Table 1) and, therefore, the latter slightly less thermally stable. The processes of thermal decomposition for 1:1 products prepared by coacervation alone and by electrosynthesis were practically identical (Fig. 3). Slightly higher water content in the electrosynthesised product might be a result of the slower and, perhaps, more ordered building up of the matrix. The result of the DTG analysis (Table 4) indicated that both preparation methods produced products of dierent macrostructure. Water in the product formed by coacervation was held more strongly than in the product prepared by electrosynthesis. It is suggested that the latter procedure provided a more compact and, possibly, a better organised matrix. TG and DTG analyses showed that the XG complexes decomposed in three steps. The third step took

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Table 4 TG and DTG analysis of G, X as well as 1:1 XG product obtained by simple coacervation and by electrosynthesis Sample Analysis TG Temperature of the eect (C) G 156.4 227.3 227.4 144.2 206.7 324.5 124.3 180.0 240.0 270.0 370.0 124.6 180.0 240.0 270.0 370.0 Associated weight lossb (%) 10.2 11.4 52.8 18.0 19.3 56.7 9.2 10.2 17.5 27.5 60.0 10.1 11.2 18.0 28.0 60.0 DTGa (C)

sules, and in application as sizes and biodegradable materials.

3. Experimental procedures
75.7 307.7c 56.5 278.9c 66.7 234.9 293.8c 337.1 60.9 237.5 296.4c 333.7

3.1. Materials X (G1253) and G (type A from porcine skin, G2500) were purchased from Sigma Chemical Co., St. Louis, MO, USA. 3.2. Electrosynthesis A 400-cm3 beaker equipped with two Pt electrodes situated at a distance of 2.5 cm from one another was used as an electrolytic cell. It was connected to a power supply (model P-3003D, Taiwan). The beaker was lled with 250 ml of aqueous solution containing 0.125 g of X and 0.125, 0.250, 0.375 g of G. These concentrations corresponded to X:G ratios of 1:1, 1:2, 1:3, 1:4 (w/w), respectively. The polymer solution was completely dissolved in distilled water under conditions of mild heating ($40 C) and stirring, then cooled to room temperature. The pH was adjusted to 9, 10 and 11 with 0.01 M NaOH. The electrosynthesis was conducted at a constant potential dierence of 12 V for 1.5 h. A layer of white, gummy product was collected from the surface of anode, centrifuged to remove liquid, and dried in vacuum at room temperature. 3.3. Sample coacervation Aqueous blends of X and G as described in Section 3.2 were adjusted to a pH 2.3 with 0.1 M hydrochloric acid whilst being agitated. The resulting precipitates were centrifuged to remove liquid, and dried in vacuum at room temperature. 3.4. Combustion analysis

1:1 GX from simple coacervation

1:1 GX from electrosynthesis

All eects were endothermic. The weight loss from the origin. c The dominating eect.
b

Fig. 3. The TG curve of the 1:1 GX product obtained by simple coacervation () and by electrosynthesis ( ).

Combustion analysis was carried out with a Perkin Elmer 2400 CHN elemental analyzer (Norwalk, CT, USA). Analyses were duplicated. They varied by up to 0.25% of recorded values. 3.5. Solubility tests The solubility of starting materials and the products in water, 5% hydrochloric acid, 5% Na2 CO3 , 0.11 M NaOH, DMSO, and 7 M aq. urea were examined at 25 C.

place at temperature above the decomposition point of X and G and this thermal stabilisation might be accounted for as a result of hydrophobic interactions. Coacervation of X with G from solutions of X and G suggested that such a process might be useful in recovery of X from aqueous solution. Moreover, the increased stability of the complexes under acidic conditions could be of practical signicance in formation of microcap-

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3.6. FTIR analysis The infrared spectra of X, G, and their complexes were run in KBr discs using a Perkin Elmer FTIR spectrometer Paragon 1000 (Norwalk, CT, USA), in the frequency range of 4000500 cm1 . Dierential spectra of XG complexes and pure X or G were also recorded. 3.7. Thermal analysis The thermal characteristics of pure X, G, and XG complexes were determined under a nitrogen stream using a Du Pont TGA 951 system (Wilmington, DE, USA) scanned from 25 to 500 C at 10 C/min.

4. Conclusions Regardless of preparation methods, X and G formed complexes with the involvement of weak electrostatic and non-electrostatic interactions between X and G. The matrix of the product formed from electrosynthesis could be more regularly in structure. The role of the electrode process carried the formation around the anode of a concentration of protons suitable for coacervation of G and X.

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