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Effect of Tetra-Alkyl Ammonium Bromide on the Rheological and Thermal Properties of Kappa-Carrageenan and Agarose Gels

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Effect of Tetra-Alkyl Ammonium Bromide on the Rheological and Thermal Properties of Kappa-Carrageenan and Agarose Gels Polymer Journal, Vol. 22, No. 11, pp 991-999 (1990) Effect of Tetra-Alkyl Ammonium Bromide on the Rheological and Thermal Properties of Kappa-Carrageenan and Agarose Gels Mineo WATASE*, Katsuyoshi NISHINARI,**·t Peter A. WILLIAMS,** and Glyn 0. PHILLIPS** * Faculty of Liberal Arts, Shizuoka University, Ohya, Shizuoka 422, Japan ** The North East Wales Institute of Higher Education, Deeside, Clwyd, CH5 4BR, UK (Received April 16, 1990) ABSTRACT: Dynamic viscoelastic measurements and differential scanning calorimetry were carried out for kappa-carrageenan gels containing tetra-alkyl ammonium bromide in order to clarify the structure and functional properties of kappa-carrageenan. The dynamic Young's modulus E' of the gels showed a maximum as a function of concentration of tetra-alkyl ammonium bromide at around 0.2 mol I- 1 . At any particular temperature E;,.., decreased in the following order: E;,..,(Me4 N-)>E;.• .(Et 4 N-)>E;..,(Pr4 N-)>E;,.• .(Bu 4 N-). The temperatures Tm of the endothermic peak accompanying the transition from gel to sol in the heating DSC curves shifted to higher temperatures with increasing salt concentration. The order of the melting tempera­ ture of these gels was the same as in E;,..,, however, the enthalpy of melting showed the reverse order. Cations with smaller dynamic hydration number noHN increase the elastic modulus and the melting temperature of kappa-carrageenan gels more than those with larger noHN· KEY WORDS Kappa-Carrageenan / Agarose /Gel/ Tetra-Alkyl Ammoni- um Bromide / Rheology / Differential Scanning Calorimetry / Dynamic hydration Number / Kappa-carrageenan and agarose are seaweed polysaccharides is considered as follows: (i) polysaccharides, and their chemical structures single or double helices are formed from resemble each other: Kappa-carrageenan con­ random coils by lowering the temperature of sists of o-galactose 4 sulfate and 3,6-anhydro­ the solution, (ii) these helices aggregate to form o-galactose, while agarose consists of o­ an ordered structure which is called the galactose and 3,6-anhydro-L-galactose. How­ junction zone or the crystalline region, and (iii) ever, their gelling properties are quite different: as a result, the three dimensional network is the gelling ability of kappa-carrageenan is formed. Therefore, these gels consists of increased by the addition of a small amount junction regions connected by long flexible of alkali metal ions, alkali earth ions 1 - 6 and chains. The junction zones are considered to guanidine hydrochloride. 7 The addition of be formed by hydrogen bonds. Kappa-carra­ these chemical reagents in excess quantity geenan contains sulfate esters, while agarose decreases the gelling ability of kappa-carra­ does not. Therefore, the above mentioned ex­ geenan. The gelling ability of agarose is not perimental findings were understood as fol­ influenced by alkali metal ions or alkali earth lows: The cations may shield the electrosta­ ions, but decreased by guanidine hydro­ tic repulsion between sulfate esters in kappa­ chloride. The gelling mechanism of these carrageenan molecules, and since the electro- t Permanent Address: National Food Research Institute, Tsukuba 305, Japan. 991 M. WATASE et a/. static repulsion inhibits the formation of helices et al. 11 observed the analogous transitions in of kappa-carrageenan molecules and their the presence of bromide and chloride co­ aggregation and tight binding, the introduction anions. The effect of alkali metal ions, 12 - 14•18 of cations promotes the aggregation of and poly-hydric alcohols15 on the gel forming kappa-carrageenan molecules, and this in­ ability ofkappa-carrageenan and agarose have creases the gelling ability. Since agarose has no been studied by rheological measurements and sulfate esters, the cations have less influence on differential scanning calorimetry (DSC). the gelling ability. This interpretation was In the present work, the effects of tetra-alkyl confirmed by the effect of guanidine hydro­ ammonium salts on the rheological and chloride (GuHCI) and urea on rheological and thermal properties of Kappa-carrageenan and thermal properties of these gels. 7 Both of these agarose gels were examined in order to compare chemical reagents are hydrogen bond breakers. with the effects of other chemical reagents on However, GuHCI is an electrolyte, and so the these properties. The gelling ability of kappa­ guanidinium ions shield the electrostatic carrageenan will be influenced by two factors: repulsion between sulfate esters in kappa­ (i) tetra-alkyl ammonium salts will play a role carrageenan as alkali metal ions or alkali earth as the electrolyte and interact with kappa­ metal ions do. Therefore, the addition of carrageenan molecules, and (ii) they will change GuHCl below a certain concentration increases the structure of water as a solvent and therefore the gelling ability of kappa-carrageenan, but it will also affect the structure of the gels. decreases the gelling ability of agarose. Since urea is not an electrolyte, it only decreases the EXPERIMENTAL gelling ability of both polysaccharides. Tetra-alkyl ammonium salts have attracted Materials much attention from many investigators Kappa-carrageenan was extracted from because of their importance in the study of Chondrus crispus as used in the previous hydrophobic interaction and the iceberg report. 7 The intrinsic viscosity in 0.01 N formation. 8 NaSCN solution at 35°Cwas 14.0 (100ml g- 1). Grasdalen and Smidsrod2 •9 reported that the The potassium content in the specimen mea­ conformational transition of kappa-carrageen­ sured by atomic absorption spectrophotom­ an molecules detected by the change of specific etry was 3.24%. Agarose was extracted from optical rotation occurred by lowering the Gelidium amansii as used in the previous temperature in tetramethyl iodide but not in report. 7 It was pretreated with 3% NaOH, and tetramethyl chloride. This result together with then washed in flowing water. The intrinsic the invariance of molecular weights of viscosity in 0.01 N NaSCN at 35°C was 3.3 tetramethyl-ammonium kappa-carrageenate in (100 ml g- 1 ). The preparation methods used N(CH3) 4 salts led them to the adoption of a in this study were the same as reported pre­ single-stranded carrageenan-iodide complex viously_ 7, 12 - 1 s rather than the double-helix structure that had Extra pure reagent grade (CH3) 4 NBr, 1 previously been proposed by Morris and Rees. (C2 H 5) 4 NBr, (C 3 H 7) 4 NBr and (C4 H 9 ) 4 NBr Norton et al. 11 also observed the order­ (Wako Pure Chemical Co., Ltd.) were used disorder transition by optical rotation mea­ without further purification. surement for tetramethyl ammonium kappa­ carrageenate in the presence of (Me)4NC1, Measurements (Me)4 NBr and (Me)4 NI. Although Grasdalen The dynamic Young's modulus E' at 2.5 Hz and Smidsrod2•9 reported that this order­ was measured at various temperatures using a disorder transition is unique to iodide, Norton Rheolograph Gel CV-100 from Toyo Seiki 992 Polym. J., Vol. 22, No. II, 1990 Carrageenan and Agarose Gels with Tetra-Alkyl Ammonium Bromide Seisakusho Ltd. The temperature was con­ experimental methods were described pre­ trolled by a silicone oil bath, and the gel sample viously .12 -15 was kept at each measurement temperature for 20 min. Differential scanning calorimetry RESULTS AND DISCUSSION (DSC) was carried out with a highly sensitive DSC SSC580 from Seiko Electronics Ltd. The Figures 1 and 2 show the dynamic Young's heating rate was fixed as 2°C min - i throughout modulus E' of2% w/w kappa-carrageenan gels the present work. The details of these as a function of concentration of tetra-alkyl 15-----------~ ~E 10 'l'E z z 10 ...0 ''o X X w w 0 0 0,5 M~NBr cone. (mol/1) Et4 NBr cone. (mol /I) Figure I. The dynamic Young's modulus E' of 2% w/w kappa-carrageenan gels with and without tetra-alkyl ammonium bromide as a function of concentration of Me4 NBr (A) and Et4 NBr (B) at various temperatures: a, 15°C; b, 25°C; c, 35°C; d, 45°C. 10 8 (D) "''E "IE z z 5 "o "o 4 X >< UJ w 0 0.5 I 0 0.2 0~ Pr4 NBr cone. (mol/1) 81J4N8r cone. (mol/ ll Figure 2. The dynamic Young's modulus E' of 2° w/w Kappa-carrageenan gels with and without tetra-alkyl ammonium bromide as a function of concentration of Pr4 NBr (C) and n-Bu4 NBr (D) at various temperatures: a, 15°C; b, 25°C; c, 35°C; d, 45°C. Polym. J., Vol. 22, No. 11, 1990 993 M. WATASE et a/. 4~-------------, 4 ~------------, 3 3 ~E "''E z z ., "'o 2 0 2 X X ' w 'w o o.5 r 1.5 0 0.7 1.4 Me.iNBr cone. (mol/1) Et4 N Br cone. ( mol /I) Figure 3. The dynamic Young's modulus E' of 3% w/w kappa-carrageenan gels with and without tetra-alkyl ammonium bromide as a function of concentration of Me4 NBr (A) and Et4 NBr (B) at various temperatures: a, 15°C; b, 25°C; c, 35°C; d, 45°C, e, 55°C. 3 2 (D) "''E 2 "''E z z "'o "'o >< X I w ' w 0 0 0.4 0.8 B1.4NBr cone. (mol/1 ) Pr4 NBr cone. (mol/1 J Figure 4. The dynamic Young's modulus E' of 3% w/w kappa-carrageenan gels with and without tetra-alkyl ammonium bromide as a function of concentration of Pr4 NBr (C) and n-Bu4 NBR (D), at various temperature: a, 15°C; b, 25°C; c, 35°C; d, 45°C; e, 55°C; f, 65°C. ammonium bromide at various temperatures. slightly to higher concentrations with increas­ E' increased sharply up to 0.l--0.2mol 1- 1 ing temperature.
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