Investigation of the rheological alterations in Iota carrageenan through the exchange of potassium with calcium ions.

Investigation of the rheological alterations in Iota carrageenan through the exchange of potassium with calcium ions. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Investigation of the rheological alterations in Iota carrageenan through the exchange of potassium with calcium ions. Shashank Kailkhura, Akanksha Bhatt, Priyank purohit This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3421472/v2 This work is licensed under a CC BY 4.0 License Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Abstract Carrageenan is a natural polymer extracted from the Red-Seaweeds that is important for the food and pharmaceutical industries. It is crucial to comprehend how the ion exchange affects the rheological properties of Iota carrageenan polymer, particular potential for gelation, which has several applications in the food and pharmaceutical industries. The current study focuses on analyzing the rheological changes brought on by the exchange of potassium ions with calcium ions in Iota carrageenan. Due to their distinct associations with the anion component, monovalent and divalent cations both have a varied impact on the gelation property of carrageenan. Polymer Science Carrageenan Polysaccharide conformational analysis cation gelation behavior Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 INTRODUCTION The gelling property of polymers and modified polymers[ 1 ], as well as other unique physicochemical qualities, have led to their employment in a variety of industries. A carrageenan, which has biological features such as antiviral, anticoagulant, immunomodulatory, and anticancer, which is derived from red algae and has a galactopyranosal residue with alpha 1–3 and beta 1–4 linkages and a sulfate group. There are fifteen different type of carrageenan categorized based on repeat of disaccharide in presence and absence of sulfate ester at free hydroxyl positions and so far only kappa, iota and lambda carrageenan has been explored consisting of 1,2 and 3 sulfate group respectively.[ 2 – 3 ]Cations play important role in gelling ability of carrageenan and from reported studies it was suggested that the double helix formation and gelatin is because of salt present with it or temperature based aggregation of helices. The gelling mechanism of carrageenan is mainly attributed to the interactions between the carrageenan polymer chains and cations in the surrounding environment. These cations can bridge the negatively charged sulfate groups on the carrageenan molecules, promoting the formation of double helices or helical junction zones. This cross-linking of carrageenan chains by cations results in the formation of a gel network (Fig. 1 ) [ 4 – 5 ] The gelation mechanism and rheological behavior of iota carrageenan has been studied by spectroscopic studies using IR, effects of monovalent and divalent on gelation property. [ 7 – 12 ]In the present research our aim is to demonstrate the vital role of cations and changing cation from K + to Ca 2+ (Mono valent to divalent) can lead to the different polymeric characters. However, with presence of specific cations like potassium ion in iota carrageenan is extensively studied in our lab and also in the research arena. In the case of K + linked carrageenan the dissociated ion of iota carrageenan gets its stable conformer in the normal water and form gel and in the hot water it forms water through staggered conformers (high energy conformers) through the di sulfate ion bridge. (Fig. 2 )[ 13 – 15 ] The conformation changes in it by initializing coil to helix transition and leads to gelation due to aggregation of these helices.[ 16 ] This study belongs to detail study about the replacement of k ions to Ca ions in carrageenan sample and observed physiochemical changes due to presence of Ca ion while comparing with potassium linked carrageenan and investigation of the molecular interactions between carrageenan and Ca + 2 ions using relevant experiments. It could be useful for the potential biological applications of Ca + 2 -linked carrageenan, such as drug delivery systems or wound healing agents.[ 17 ] MATERIAL AND METHOD The laboratory-grade Calcium Chloride, sliver nitrate and solvents like ethanol were bought from Aldrich (Merck), while compounds like K salt of iota carrageenan were procured from marine hydrocolloids (cochin) with a respectable purity profile (WATERGEL GU – 8684 is a pure refined Iota carrageenan). Glassware including beakers, glass rods, measuring cylinder, funnel and micropipettes were employed, along with equipment like magnetic stirrers (Adarsh Model Number (9001 − 2815)), Ultrasonicator [Zhart India (2018/108)], weighing machines (Electronic compact scale (SF-400C)).The research will involve a combination of experimental techniques, including spectroscopy (e.g., FTIR,), rheological analysis, and biological assays. The study will be conducted through a series of controlled experiments with varying concentrations of Ca + 2 ions and carrageenan, followed by comprehensive data analysis and interpretation. The various procedures were carried out in accordance with the information provided below with the relevant detail. 1. Reaction with CaCl 2 Here Reaction of potassium linked carrageenan is done with calcium chloride to get the Ca linked carrageenan by ion exchange method. Carrageenan's interactions with calcium ions (Ca + 2 ), which play a crucial role in its gelation and other functional properties. [ 18 – 19 ] Step I – Carrageenan's interactions with calcium ions (Ca + 2 ) was done by using calcium chloride and sliver nitrate. The procedure followed by adding 500 mg calcium chloride in 20 ml water by continuously stirring. After 10 minutes 3 ml of AgNO 3 was added to this solution drop wise, resulting settle down of AgCl 2 ppt. So separation of Ca containing layer was done by pouring it into another beaker carefully. Step II - In another beaker 500 mg carrageenan was mixed with ethanol (40 ml) and further with water (30 ml) and stir for 30 minutes Finally, combine the resulting solution obtained from Step I and II 2. Confirmation of Ca 2+ ion linkage: The NaOH was added to the water carrageenan (Ca linked carrageenan) solution to see the interference of OH ions to form calcium hydroxide on reaction and obtained in the form of ppt. Instrumental method to confirm the Ca 2+ ion : IR : The FTIR equipment utilized in this study was the Spectrum 2 model, with a serial number of 89258. The software employed for data analysis was NIOS2. KBr palates were employed as the sampling medium for the chemicals under investigation. MS : The instrument under consideration is a mass spectrometer manufactured by Shimadzu in Japan. The instrument utilized for the analysis was the Shimadzu QP 2010 Ultra Mass Spectrometry (MS) system. The apparatus is outfitted with a mass spectrometer (MS), an electron capture detector (ECD), and a flame ionization detector (FID). The aforementioned apparatus was employed for the purpose of analysing Mass spectra. Viscosity: The rotational viscometer was used to calculate the viscosity; the temperature was kept constant at 30 o C and 30 RPM during the process; additionally, the spindle L2 was utilized to analyze the sample's viscosity, and the regular gel and nano gel were examined, to check the easy applicability for topical route, Conductivity Test The conductivity of both nano and simple gel was measured by using conductivity meter, which was calibrated through distilled water solution. Computational method The bond length of the K + and Ca 2+ ion with carrageenan was calculated through chem 3D 16 ultra commercially available software. The energy minimization and bond were normalized through MM2 method and the ionic interaction distance was calculated. RESULT AND DISCUSSION The confirmation test of the solid product was done using 0.01N NaOH and the confirmation of Ca 2+ linkage was done, When NaOH reacts with Ca²⁺ ions, it forms calcium hydroxide (Ca (OH)₂), which is sparingly soluble in water and forms a white precipitate. The reaction is represented as follows: Ca²⁺ + 2NaOH ---Ca (OH)₂↓ + 2Na⁺ The formation of this white precipitate (Fig. 3 ) is a positive indication of the presence of calcium ions. Identification of Ca 2+ ion attachment through IR from K + linked carrageenan. Since calcium ions (Ca 2+ ) do not have any identifiable IR absorption bands, identifying Ca 2+ ion attachment using IR spectroscopy can be difficult. However, Ca 2+ ions have distinctive IR absorption bands that is utilized to distinguish them when they bind with another compounds. The comparison of the IR of K + and Ca 2+ ion attachment using IR spectroscopy have been done through some of the IR peaks given in the Fig. 4 . The peak observed at 1644 cm-1, corresponding to the O-H bending "deformation," provides insights into the presence of water bonded to the polymer. This peak has shifted to a higher infrared (IR) value and exhibits a higher intensity. These observations suggest that the K + ions have been replaced, likely due to the smaller size of Ca 2+ ions, which results in a stronger attraction towards water and sulfate ions. The O-S stretching frequency in the presence of Ca 2+ ions is measured at 1288 cm-1 with an absorbance of 85%. In contrast, when K + ions are present, the O-S stretching frequency is observed at 1304 cm − 1 with an absorbance of 70%.[ 20 ] Additionally, this observation indicates the presence of a strong interaction and an accompanying decrease in bond length. This phenomenon results in a drop in the S-O frequency, indicating that the S-O–Ca 2+ contacts are stronger compared to the S-O----K + interaction. The observed displacement of the peak and its corresponding intensity provides confirmation of the substitution of K + ions with Ca 2+ ions in the carrageenan, while leaving the fingerprint region unaffected (721cm − 1 , 1456 − 1300 cm − 1 two intense peaks of SO 2 , 1280–1300 cm − 1 of S-O bond.) [ 21 – 23 ] Mass spectroscopy. The mass spectra provide compelling evidence for the substitution of K + ions with Ca 2+ ions in the iota carrageenan. (Fig. 5 ) The presence of the fragment peak [136] + (477 − 341) provides confirmation of the existence of the [SO 4 Ca] + fragment. Similarly, the presence of the [154] + (221 − 77) fragment verifies the presence of [SO4Ca 2+ . H 2 O] + , indicating the polymer's association with water. Notably, the intensity of this peak is particularly high in the case of Ca 2+ due to its strong affinity for water and its network. Furthermore, the observed peak at 96 in the mass spectrum provides confirmation of the presence of the [SO 4 ] + fragment. The presence of [120] + (SO3Ca) + and the inference of (133 − 77) = [56] + provide evidence supporting the existence of the [OCa 2+ ] fragment. This comprehensive analysis verifies the identification of carrageenan as the ion carrying the Ca 2+ ion. Furthermore, the characteristic base peak observed at [207] + provides additional confirmation of the carrageenan skeleton. Optimization of the solvent : The choice of solvent can influence how carrageenan and interacts with other components. Common solvents include water and organic solvent were screened to analyse the gelling properties of carrageenan, The carrageenan with divalent cations like Ca²⁺ and its status in different solvent were noted as depicted in the below Table 1 . Table 1 Solvent Screening and Solubility Profile of carrageenan. S. No. Solvents Observation 1 Ethyl Acetate* Not Forming Gel and Insoluble 2 Ethanol Not Forming Gel and Insoluble 3 Dichloromethane (DCM) Not Forming Gel and Insoluble 4 N, N, dimethyl Formamide (DMF) * Not Forming Gel and partially soluble 5 Acetone Not Forming Gel & Insoluble 6 Toluene* Not Forming Gel and Insoluble 7 Acetone Not Forming Gel and Insoluble 8 Dimethyl sulfoxide (DMSO)* Not Forming Gel and Insoluble 9 Methanol Not Forming Gel and Insoluble 10 Acetonitrile* Not Forming Gel and Insoluble 11 Water Not Forming Gel and Insoluble 12 Water (Hot) Gel formation * Temperature was also enhanced till boiling point and similar observation was observed. The solubility of the Ca + 2 Carrageenan was tested using polar protic (Acetone, Alcohols), aprotic (DMF), high boiling polar solvent (DMSO), and non-polar (toluene) solvents; however, none of the solvents were successful in producing the carrageenan solution phase, Table 1 (Entry 1–10), even after raising the temperature to the boiling point of the corresponding solvent. It was anticipated that the polymer would remain in solution phase following dissociation, which was only made possible in water due to the solvent's high dissociation capacity. After screening with organic solvents, water (Table 1 entry 11) was utilized as an aqueous solvent. Surprisingly, it wasn't solvable, and the outcomes from entries 1 through 11 were comparable as before. The temperature of the carrageenan solution in water was gradually raised until it took the form of a gel; however, in the case of K + , the gel form was with water at a normal temperature and the solution phase was with a high temperature; the opposite result with Ca 2+ provided a reason to further optimize the temperature effect to see how the behaviour changed as the water temperature changed. Table :2 Effect of temperature in the water and carrageenan system: Ice-Water Normal Water Hot water 48-80 o C High Temperature 80 o C Not Forming Gel, Insoluble Not Forming Gel, Insoluble Gel formation Viscous solution, soluble The temperature-based analysis shows the pattern of Gel-sol property after the heating, however in case of K + the sol-Gel conversion was taking place with respect to the heating. The further investigation of the ion- effect was caried out, by keeping in the mind the Ca 2+ is smaller (2.4 Å) than the potassium (2.7Å) and the computational based study regarding the distance between sulphate ion and K + and Ca 2+ as depicted in Fig. 6 . The distance between Ca 2+ and carrageenan was less than the K + , it was also clue regarding the more attraction or more bond dissociation energy with the Ca 2+ ion. It was anticipated that the cold/normal water does not converts in to the ionic form, however the high temperature when matches with the dissociation energy gets in to the dissociation for and forms gel with stable 1 C 4 conformers for the formation of gel. It was also interesting to see that heating more than 80 o Cwas converting Gel to solution, because of its conformational change from 4 C 1 , A higher energy state for solution as it is related with the reported study regarding the role of conformational changes and its effect on phase, as depicted in the below Fig. 6 . Moreover, the network with H 2 O which was observe in the IR and Mass also suggest the close linkage of the network in undissociated form, which does not get ionized in the normal temperature so the higher temperature is require to form the gel in with stable 1 C 4 conformer as depicted in Fig. 7 . However, in the case of K + the bond attraction was less so it dissociated in normal water and forms gel, and in case of the hot water it forms solution because of the conformational change to 4 C 1 as solution phase. Figure 8 The dissociation theory for Ca 2+ was further analyse through the ionic content in the solution with respect to the temperature. The carrageenan and water solution were reacted with the NaOH solution in each temperature and the pattern of the reactivity was found different as per given Table 3 Table 3 Reaction of free calcium ion with NaOH in different temperature. Ice-Water Normal Water Hot water 48-80 o C High Temperature 80 o C After the addition of NaOH, the solution turns viscous. After the addition of NaOH, the solution turns more viscous After the addition of NaOH solution, it does not give any change After the addition of NaOH solution, it does not give any change Very light Precipitate forms after 15–20 mints with less yield Light precipitate forms in 15 minutes Precipitate in few minutes with high yield Precipitate in few minutes with high yield The reactivity pattern with NaOH clearly represent the availability of free calcium ion present in the solution, it also confirms the dissociation of the Ca 2+ is also increasing with high temperature and justifying our plausible mechanism regarding the undissociated form in cold water of Ca-linked carrageenan. The ionic content in hot water was also confirm by the conductivity as the io gets dissociated the conductivity increase and the similar result obtained in the test, and the gelling property correlation with ionized and unionized form was confirm through the experiment. Conductivity based on the principle by measuring the electric current flows in electrons present in the solution means if the solution is higher in ion concentration shows reading towards higher, which is observed in our result, as ca ion are in dissociated form in hot water so showing conductivity more than cold and normal water. However. The conductivity of the K + carrageenan remains same in all the condition, so the dissociation was same in the all cases, while with the Ca 2+ it increases with the temperature. Table 4 conductivity of the water carrageenan solution in different temperatures. Ca CG with water at different temperature Readings Hot water (50-100 0 C) 0.59–0.64 Water (RT) 0.01 Cold water (8 0 C) Normal 0.01 The investigation into the Ca 2+ linked carrageenan and its temperature-dependent role is supported by a number of chemical and experimental evidence, and it is suggested that because the Ca 2+ linked carrageenan has a higher dissociation energy than the K + linked carrageenan, it forms gel in hot water and is insoluble in cold water, supporting the role of cations and conformational changes. Viscosity: The viscosity of the K + linked gel was found as 649 centipoise and the gel with Ca 2+ has 728, (Table 5 ) which also justifies the higher cross linking with helical-to-helical linkage to form more stiff and stable gel. The intermolecular interactions (Fig. 9 ) were also observed by IR through the OH deformation peak, so the more interactions get more stiff gel. Table 5 Viscosity results of nano and simple gel Type of Gel Viscosity K + Carrageenan gel 649 Ca + Carrageenan gel 728 CONCLUSION Iota carrageenan plays important role either in food industry or for medical purposes due to their numerous applications. The effect of cations plays crucial role in case of carrageenan or we can say changes in their physiochemical behavior, that synergizes its effect and application. Also, the type of cation bivalent, monovalent is also responsible for it, affected the gelling property of carrageenan, with calcium here we find not forming gel in normal water while comparing to potassium because of the requirement of high energy for dissociation. Our study finds out the entire parameter proofing the higher energy required for Ca 2+ to dissociate and so forming gel in hot water. DECLARATIONS Acknowledge: All authors express their gratitude to Graphic Era Hill University for providing the initial funding to start the project work. Funding Declaration : Not any Conflict of Interest- Not any REFERENCES Purohit P, Bhatt A, Mittal RK, Abdellattif MH, Farghaly TA. Polymer Grafting and its chemical reactions. Frontiers in Bioengineering and Biotechnology. 2023 Jan 11;10:1044927. Evageliou VI, Ryan PM, Morris ER. Effect of monovalent cations on calcium-induced assemblies of kappa carrageenan. Food Hydrocolloids. 2019 Jan 1;86:141-5. Morris ER, Rees DA, Robinson G. Cation-specific aggregation of carrageenan helices: domain model of polymer gel structure. Journal of molecular biology. 1980 Apr 5;138(2):349-62. Janaswamy S, Chandrasekaran R. Heterogeneity in iota-carrageenan molecular structure: insights for polymorph II→ III transition in the presence of calcium ions. Carbohydrate Research. 2008 Feb 4;343(2):364-73. Morris ER, Rees DA, Robinson G. Cation-specific aggregation of carrageenan helices: domain model of polymer gel structure. Journal of molecular biology. 1980 Apr 5;138(2):349-62. van de Velde F, Rollema HS, Grinberg NV, Burova TV, Grinberg VY, Tromp RH. Coil–helix transition of ι‐carrageenan as a function of chain regularity. Biopolymers: Original Research on Biomolecules. 2002 Nov 15;65(4):299-312. Robal M, Brenner T, Matsukawa S, Ogawa H, Truus K, Rudolph B, Tuvikene R. Monocationic salts of carrageenans: Preparation and physico-chemical properties. Food Hydrocolloids. 2017 Feb 1;63:656-67. MacArtain P, Jacquier JC, Dawson KA. Physical characteristics of calcium induced κ-carrageenan networks. Carbohydrate Polymers. 2003 Sep 1;53(4):395-400. Van de Velde F, Knutsen SH, Usov AI, Rollema HS, Cerezo AS. 1H and 13C high resolution NMR spectroscopy of carrageenans: application in research and industry. Trends in Food Science & Technology. 2002 Mar 1;13(3):73-92. Leal D, Matsuhiro B, Rossi M, Caruso F. FT-IR spectra of alginic acid block fractions in three species of brown seaweeds. Carbohydrate research. 2008 Feb 4;343(2):308-16. Bongaerts K, Paoletti S, Denef B, Vanneste K, Cuppo F, Reynaers H. Light scattering investigation of ι-carrageenan aqueous solutions. Concentration dependence of association. Macromolecules. 2000 Nov 14;33(23):8709-19. Janaswamy S, Chandrasekaran R. Effect of calcium ions on the organization of iota-carrageenan helices: an X-ray investigation. Carbohydrate Research. 2002 Mar 15;337(6):523-35. Fontes-Candia C, Ström A, Lopez-Sanchez P, López-Rubio A, Martínez-Sanz M. Rheological and structural characterization of carrageenan emulsion gels. Algal Research. 2020 May 1;47:101873. Nickerson MT, Darvesh R, Paulson AT. Formation of Calcium‐Mediated Junction Zones at the Onset of the Sol‐Gel Transition of Commercial κ‐Carrageenan Solutions. Journal of food science. 2010 Apr;75(3):E153-6. Tarı Ö, Pekcan Ö. Comparison of cation effects on phase transitions of kappa and iota carrageenan. e-Polymers. 2010 Dec 1;10(1):083. Viebke C, Piculell L, Nilsson S. On the mechanism of gelation of helix-forming biopolymers. Macromolecules. 1994 Jul;27(15):4160-6. Neamtu B, Barbu A, Negrea MO, Berghea-Neamțu CȘ, Popescu D, Zăhan M, Mireșan V. Carrageenan-based compounds as wound healing materials. International Journal of Molecular Sciences. 2022 Aug 14;23(16):9117. Montero P, Pérez-Mateos M. Effects of Na+, K+ and Ca2+ on gels formed from fish mince containing a carrageenan or alginate. Food Hydrocolloids. 2002 Jul 1;16(4):375-85. Bui VT, Nguyen BT, Nicolai T, Renou F. Mixed iota and kappa carrageenan gels in the presence of both calcium and potassium ions. Carbohydrate polymers. 2019 Nov 1;223:115107. Seki T, Chiang KY, Yu CC, Yu X, Okuno M, Hunger J, Nagata Y, Bonn M. The bending mode of water: A powerful probe for hydrogen bond structure of aqueous systems. The journal of physical chemistry letters. 2020 Sep 15;11(19):8459-69. Ghani NA, Othaman R, Ahmad A, Anuar FH, Hassan NH. Impact of purification on iota carrageenan as solid polymer electrolyte. Arabian journal of chemistry. 2019 Mar 1;12(3):370-6. Rajasulochana N, Gunasekaran S. Analysis on the seasonal variations in carrageenans of Hypnea flagelliformis and Sarconema filiforme by FTIR spectroscopy. Asian Journal of Chemistry. 2009 Jul 1;21(6):4547. Jumaah FN, Mobarak NN, Ahmad A, Ghani MA, Rahman MY. Derivative of iota-carrageenan as solid polymer electrolyte. Ionics. 2015 May;21:1311-20. Additional Declarations The authors declare no competing interests. Supplementary Files SUPPLIMENTRYFILE.docx SI Cite Share Download PDF Status: Posted Version 2 posted You are reading this latest preprint version Show more versions Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3421472","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":239178497,"identity":"4360fc10-0e34-411f-8cc5-a002ae2db424","order_by":0,"name":"Shashank Kailkhura","email":"","orcid":"https://orcid.org/0009-0000-2410-7951","institution":"Graphic Era Hill University","correspondingAuthor":false,"prefix":"","firstName":"Shashank","middleName":"","lastName":"Kailkhura","suffix":""},{"id":239178498,"identity":"0130cb41-b353-420c-8de5-967698e854a8","order_by":1,"name":"Akanksha Bhatt","email":"","orcid":"","institution":"Graphic Era Hill University","correspondingAuthor":false,"prefix":"","firstName":"Akanksha","middleName":"","lastName":"Bhatt","suffix":""},{"id":239178499,"identity":"ff7559cd-a855-4c13-b759-2035a602ec4c","order_by":2,"name":"Priyank purohit","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/ElEQVRIiWNgGAWjYFACxgYgkcDAwAziVAAxM3MDKVrOgLQwEtICBglQ7W1wQ3ADc+nDbRI/d6QxmLPzHnxcOa82mr8dqOVHxTacWiz7Etske8/kMFg28yUbnt12PHfGYcYGxp4zt3FqMTjD2CbB21bBYHCYx0yycdux3AagFmbGNvxaJP9CtJj/bJxzLHc+MVqkedtywLYwNjbU5G4gpMWyh7HZWrYtjcfgMF+yZMOxA7kbgVoO4vOLOQ/7w5tv25LlDM6fPfixoaYud975wwcf/KjA4zAGBhYJIM0DRgwMh8GiB3Cqh2hh/gBhgrXU4VM8CkbBKBgFIxQAAIKoWI/AMefrAAAAAElFTkSuQmCC","orcid":"https://orcid.org/0000-0002-1011-4736","institution":"Graphic Era Hill University","correspondingAuthor":true,"prefix":"","firstName":"Priyank","middleName":"","lastName":"purohit","suffix":""}],"badges":[],"createdAt":"2023-10-08 13:59:05","currentVersionCode":2,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-3421472/v2","doiUrl":"https://doi.org/10.21203/rs.3.rs-3421472/v2","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":49142815,"identity":"21928c87-d214-4f85-957a-c8ac16bd45f6","added_by":"auto","created_at":"2024-01-03 18:45:39","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":232650,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eCation’s capability to enhance gelation based on changes in their helical orientation [6]\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/ae9525e63cd35f08f6c7b300.jpg"},{"id":49142816,"identity":"9905b89d-333a-4761-a59c-5cc9a2eb0bd9","added_by":"auto","created_at":"2024-01-03 18:45:39","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":123194,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eConformational change of carrageenan in the cold and hot water\u003c/strong\u003e.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/a4bd3cddf837cf396c60c7cf.jpg"},{"id":49143608,"identity":"a874e33a-ab41-4e9f-be6d-d9ce3b51c494","added_by":"auto","created_at":"2024-01-03 18:53:39","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":189318,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eConfirmation of the Ca\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2+\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e Linkage with iota carrageenan.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/55e66e82883b35d6524cfaa8.jpg"},{"id":49142818,"identity":"7c889c8c-570e-466f-98e5-e6f492386a7b","added_by":"auto","created_at":"2024-01-03 18:45:39","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":607068,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eInterpretation of Ca+ and K+ ion linked carrageenan with IR.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/b688207afcc46211b5a45d3d.png"},{"id":49143610,"identity":"36b4b79f-886c-4f4f-bf57-381a60c9337e","added_by":"auto","created_at":"2024-01-03 18:53:39","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":252758,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eMass spectra of the Ca\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2+\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e lined carrageenan.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/69b0203530b72e20c475aae1.png"},{"id":49144366,"identity":"eecdf22e-6e5f-4b26-9045-974497e6946d","added_by":"auto","created_at":"2024-01-03 19:01:39","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":212926,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEstimation of the ionic bond length with K\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e and Ca\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e+2\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003e ions.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/75acb2f4ea83d860be1db38c.png"},{"id":49143609,"identity":"e5cae9dc-d7d4-448a-b709-8c075c46334a","added_by":"auto","created_at":"2024-01-03 18:53:39","extension":"jpg","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":320203,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of temperature in the undissociated and dissociated \u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/sub\u003e\u003cstrong\u003e and \u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e4\u003c/strong\u003e\u003c/sup\u003e\u003cstrong\u003eC\u003c/strong\u003e\u003csub\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003c/sub\u003e\u003cstrong\u003e conformers\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"7.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/4a1c23bad7461ee1d425195b.jpg"},{"id":49142824,"identity":"96899c34-579a-4132-8763-4d94695b197c","added_by":"auto","created_at":"2024-01-03 18:45:39","extension":"jpg","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":234000,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of temperature on K-linked carrageenan.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"8.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/d7dcbaa900cd0bc97c196479.jpg"},{"id":49142821,"identity":"dd8cf3df-9e62-40e6-943b-bc2fa50e01d7","added_by":"auto","created_at":"2024-01-03 18:45:39","extension":"jpg","order_by":9,"title":"Figure 9","display":"","copyAsset":false,"role":"figure","size":81287,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eGel with Ca2+ ion and Water intermolecular interaction\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"9.jpg","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/6072da7e8c446ea88f6c03be.jpg"},{"id":49144482,"identity":"4c4b6ca6-1c2c-46e2-9cf4-0623bf39d51b","added_by":"auto","created_at":"2024-01-03 19:09:40","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1850950,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/928f11d4-f94c-4b07-93c0-8d5ff17fb018.pdf"},{"id":49143607,"identity":"47d2f1b8-d665-4e2e-a449-4b17827f7f42","added_by":"auto","created_at":"2024-01-03 18:53:39","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":674181,"visible":true,"origin":"","legend":"\u003cp\u003eSI\u003c/p\u003e","description":"","filename":"SUPPLIMENTRYFILE.docx","url":"https://assets-eu.researchsquare.com/files/rs-3421472/v2/fd20cab1be58dbeec25d7e16.docx"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eInvestigation of the rheological alterations in Iota carrageenan through the exchange of potassium with calcium ions.\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eThe gelling property of polymers and modified polymers[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e], as well as other unique physicochemical qualities, have led to their employment in a variety of industries. A carrageenan, which has biological features such as antiviral, anticoagulant, immunomodulatory, and anticancer, which is derived from red algae and has a galactopyranosal residue with alpha 1\u0026ndash;3 and beta 1\u0026ndash;4 linkages and a sulfate group. There are fifteen different type of carrageenan categorized based on repeat of disaccharide in presence and absence of sulfate ester at free hydroxyl positions and so far only kappa, iota and lambda carrageenan has been explored consisting of 1,2 and 3 sulfate group respectively.[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]Cations play important role in gelling ability of carrageenan and from reported studies it was suggested that the double helix formation and gelatin is because of salt present with it or temperature based aggregation of helices. The gelling mechanism of carrageenan is mainly attributed to the interactions between the carrageenan polymer chains and cations in the surrounding environment. These cations can bridge the negatively charged sulfate groups on the carrageenan molecules, promoting the formation of double helices or helical junction zones. This cross-linking of carrageenan chains by cations results in the formation of a gel network (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e) [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/p\u003e \u003cp\u003eThe gelation mechanism and rheological behavior of iota carrageenan has been studied by spectroscopic studies using IR, effects of monovalent and divalent on gelation property. [\u003cspan additionalcitationids=\"CR8 CR9 CR10 CR11\" citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]In the present research our aim is to demonstrate the vital role of cations and changing cation from K\u003csup\u003e+\u003c/sup\u003e to Ca\u003csup\u003e2+\u003c/sup\u003e (Mono valent to divalent) can lead to the different polymeric characters. However, with presence of specific cations like potassium ion in iota carrageenan is extensively studied in our lab and also in the research arena. In the case of K\u003csup\u003e+\u003c/sup\u003e linked carrageenan the dissociated ion of iota carrageenan gets its stable conformer in the normal water and form gel and in the hot water it forms water through staggered conformers (high energy conformers) through the di sulfate ion bridge. (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e)[\u003cspan additionalcitationids=\"CR14\" citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]\u003c/p\u003e \u003cp\u003eThe conformation changes in it by initializing coil to helix transition and leads to gelation due to aggregation of these helices.[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] This study belongs to detail study about the replacement of k ions to Ca ions in carrageenan sample and observed physiochemical changes due to presence of Ca ion while comparing with potassium linked carrageenan and investigation of the molecular interactions between carrageenan and Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e ions using relevant experiments. It could be useful for the potential biological applications of Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e-linked carrageenan, such as drug delivery systems or wound healing agents.[\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]\u003c/p\u003e"},{"header":"MATERIAL AND METHOD","content":"\u003cp\u003eThe laboratory-grade Calcium Chloride, sliver nitrate and solvents like ethanol were bought from Aldrich (Merck), while compounds like K salt of iota carrageenan were procured from marine hydrocolloids (cochin) with a respectable purity profile (WATERGEL GU \u0026ndash; 8684 is a pure refined Iota carrageenan). Glassware including beakers, glass rods, measuring cylinder, funnel and micropipettes were employed, along with equipment like magnetic stirrers (Adarsh Model Number (9001\u0026thinsp;\u0026minus;\u0026thinsp;2815)), Ultrasonicator [Zhart India (2018/108)], weighing machines (Electronic compact scale (SF-400C)).The research will involve a combination of experimental techniques, including spectroscopy (e.g., FTIR,), rheological analysis, and biological assays. The study will be conducted through a series of controlled experiments with varying concentrations of Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e ions and carrageenan, followed by comprehensive data analysis and interpretation. The various procedures were carried out in accordance with the information provided below with the relevant detail.\u003c/p\u003e\n\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\n \u003ch2\u003e1. Reaction with CaCl\u003csub\u003e2\u003c/sub\u003e\u003c/h2\u003e\n \u003cp\u003eHere Reaction of potassium linked carrageenan is done with calcium chloride to get the Ca linked carrageenan by ion exchange method. Carrageenan\u0026apos;s interactions with calcium ions (Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e), which play a crucial role in its gelation and other functional properties. [\u003cspan class=\"CitationRef\"\u003e18\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eStep I \u0026ndash;\u003c/strong\u003eCarrageenan\u0026apos;s interactions with calcium ions (Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e) was done by using calcium chloride and sliver nitrate. The procedure followed by adding 500 mg calcium chloride in 20 ml water by continuously stirring. After 10 minutes 3 ml of AgNO\u003csub\u003e3\u003c/sub\u003e was added to this solution drop wise, resulting settle down of AgCl\u003csub\u003e2\u003c/sub\u003e ppt. So separation of Ca containing layer was done by pouring it into another beaker carefully.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eStep II\u003c/strong\u003e- In another beaker 500 mg carrageenan was mixed with ethanol (40 ml) and further with water (30 ml) and stir for 30 minutes\u003c/p\u003e\n \u003cp\u003eFinally, combine the resulting solution obtained from Step I and II\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec4\" class=\"Section2\"\u003e\n \u003ch2\u003e2. Confirmation of Ca\u003csup\u003e2+\u003c/sup\u003e ion linkage:\u003c/h2\u003e\n \u003cp\u003eThe NaOH was added to the water carrageenan (Ca linked carrageenan) solution to see the interference of OH ions to form calcium hydroxide on reaction and obtained in the form of ppt.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eInstrumental method to confirm the Ca\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2+\u003c/strong\u003e\u003c/sup\u003e \u003cstrong\u003eion\u003c/strong\u003e:\u003c/p\u003e\n \u003col style=\"list-style-type: lower-alpha;\"\u003e\n \u003cli\u003e\n \u003cp\u003e\u003cstrong\u003eIR\u003c/strong\u003e: The FTIR equipment utilized in this study was the Spectrum 2 model, with a serial number of 89258. The software employed for data analysis was NIOS2. KBr palates were employed as the sampling medium for the chemicals under investigation.\u003c/p\u003e\n \u003c/li\u003e\n \u003cli\u003e\n \u003cp\u003e\u003cstrong\u003eMS\u003c/strong\u003e: The instrument under consideration is a mass spectrometer manufactured by Shimadzu in Japan. The instrument utilized for the analysis was the Shimadzu QP 2010 Ultra Mass Spectrometry (MS) system. The apparatus is outfitted with a mass spectrometer (MS), an electron capture detector (ECD), and a flame ionization detector (FID). The aforementioned apparatus was employed for the purpose of analysing Mass spectra.\u003c/p\u003e\n \u003c/li\u003e\n \u003c/ol\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec5\" class=\"Section2\"\u003e\n \u003ch2\u003eViscosity:\u003c/h2\u003e\n \u003cp\u003eThe rotational viscometer was used to calculate the viscosity; the temperature was kept constant at 30\u003csup\u003eo\u003c/sup\u003eC and 30 RPM during the process; additionally, the spindle L2 was utilized to analyze the sample\u0026apos;s viscosity, and the regular gel and nano gel were examined, to check the easy applicability for topical route,\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec6\" class=\"Section2\"\u003e\n \u003ch2\u003eConductivity Test\u003c/h2\u003e\n \u003cp\u003eThe conductivity of both nano and simple gel was measured by using conductivity meter, which was calibrated through distilled water solution.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eComputational method\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe bond length of the K\u003csup\u003e+\u003c/sup\u003e and Ca\u003csup\u003e2+\u003c/sup\u003e ion with carrageenan was calculated through chem 3D 16 ultra commercially available software. The energy minimization and bond were normalized through MM2 method and the ionic interaction distance was calculated.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"RESULT AND DISCUSSION","content":"\u003cp\u003eThe confirmation test of the solid product was done using 0.01N NaOH and the confirmation of Ca\u003csup\u003e2+\u003c/sup\u003elinkage was done, When NaOH reacts with Ca\u0026sup2;⁺ ions, it forms calcium hydroxide (Ca (OH)₂), which is sparingly soluble in water and forms a white precipitate. The reaction is represented as follows:\u003c/p\u003e\n\u003cdiv class=\"BlockQuote\"\u003e\n \u003cp\u003eCa\u0026sup2;⁺ + 2NaOH ---Ca (OH)₂\u0026darr; + 2Na⁺\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eThe formation of this white precipitate (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e) is a positive indication of the presence of calcium ions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eIdentification of Ca\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e2+\u003c/strong\u003e\u003c/sup\u003e \u003cstrong\u003eion attachment through IR from K\u003c/strong\u003e\u003csup\u003e\u003cstrong\u003e+\u003c/strong\u003e\u003c/sup\u003e \u003cstrong\u003elinked carrageenan.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSince calcium ions (Ca\u003csup\u003e2+\u003c/sup\u003e) do not have any identifiable IR absorption bands, identifying Ca\u003csup\u003e2+\u003c/sup\u003e ion attachment using IR spectroscopy can be difficult. However, Ca\u003csup\u003e2+\u003c/sup\u003e ions have distinctive IR absorption bands that is utilized to distinguish them when they bind with another compounds. The comparison of the IR of K\u003csup\u003e+\u003c/sup\u003e and Ca\u003csup\u003e2+\u003c/sup\u003eion attachment using IR spectroscopy have been done through some of the IR peaks given in the Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e.\u003c/p\u003e\n\u003cp\u003eThe peak observed at 1644 cm-1, corresponding to the O-H bending \u0026quot;deformation,\u0026quot; provides insights into the presence of water bonded to the polymer. This peak has shifted to a higher infrared (IR) value and exhibits a higher intensity. These observations suggest that the K\u003csup\u003e+\u003c/sup\u003e ions have been replaced, likely due to the smaller size of Ca\u003csup\u003e2+\u003c/sup\u003e ions, which results in a stronger attraction towards water and sulfate ions. The O-S stretching frequency in the presence of Ca\u003csup\u003e2+\u003c/sup\u003e ions is measured at 1288 cm-1 with an absorbance of 85%. In contrast, when K\u0026thinsp;+\u0026thinsp;ions are present, the O-S stretching frequency is observed at 1304 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e with an absorbance of 70%.[\u003cspan class=\"CitationRef\"\u003e20\u003c/span\u003e] Additionally, this observation indicates the presence of a strong interaction and an accompanying decrease in bond length. This phenomenon results in a drop in the S-O frequency, indicating that the S-O\u0026ndash;Ca\u003csup\u003e2+\u003c/sup\u003e contacts are stronger compared to the S-O----K\u003csup\u003e+\u003c/sup\u003e interaction. The observed displacement of the peak and its corresponding intensity provides confirmation of the substitution of K\u003csup\u003e+\u003c/sup\u003e ions with Ca\u003csup\u003e2+\u003c/sup\u003e ions in the carrageenan, while leaving the fingerprint region unaffected (721cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e, 1456\u0026thinsp;\u0026minus;\u0026thinsp;1300 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e two intense peaks of SO\u003csub\u003e2\u003c/sub\u003e, 1280\u0026ndash;1300 cm\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003eof S-O bond.) [\u003cspan class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan class=\"CitationRef\"\u003e23\u003c/span\u003e]\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMass spectroscopy.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mass spectra provide compelling evidence for the substitution of K\u003csup\u003e+\u003c/sup\u003e ions with Ca\u003csup\u003e2+\u003c/sup\u003e ions in the iota carrageenan. (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e) The presence of the fragment peak [136]\u003csup\u003e+\u003c/sup\u003e(477\u0026thinsp;\u0026minus;\u0026thinsp;341) provides confirmation of the existence of the [SO\u003csub\u003e4\u003c/sub\u003eCa]\u003csup\u003e+\u003c/sup\u003e fragment. Similarly, the presence of the [154]\u003csup\u003e+\u003c/sup\u003e (221\u0026thinsp;\u0026minus;\u0026thinsp;77) fragment verifies the presence of [SO4Ca\u003csup\u003e2+\u003c/sup\u003e. H\u003csub\u003e2\u003c/sub\u003eO]\u003csup\u003e+\u003c/sup\u003e, indicating the polymer\u0026apos;s association with water. Notably, the intensity of this peak is particularly high in the case of Ca\u003csup\u003e2+\u003c/sup\u003e due to its strong affinity for water and its network. Furthermore, the observed peak at 96 in the mass spectrum provides confirmation of the presence of the [SO\u003csub\u003e4\u003c/sub\u003e]\u003csup\u003e+\u003c/sup\u003e fragment. The presence of [120]\u003csup\u003e+\u003c/sup\u003e (SO3Ca)\u003csup\u003e+\u003c/sup\u003e and the inference of (133\u0026thinsp;\u0026minus;\u0026thinsp;77) = [56]\u003csup\u003e+\u003c/sup\u003e provide evidence supporting the existence of the [OCa\u003csup\u003e2+\u003c/sup\u003e] fragment. This comprehensive analysis verifies the identification of carrageenan as the ion carrying the Ca\u003csup\u003e2+\u003c/sup\u003e ion. Furthermore, the characteristic base peak observed at [207]\u003csup\u003e+\u003c/sup\u003e provides additional confirmation of the carrageenan skeleton.\u003c/p\u003e\n\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\n \u003ch2\u003e\u003cstrong\u003eOptimization of the solvent\u003c/strong\u003e:\u003c/h2\u003e\n \u003cp\u003eThe choice of solvent can influence how carrageenan and interacts with other components. Common solvents include water and organic solvent were screened to analyse the gelling properties of carrageenan, The carrageenan with divalent cations like Ca\u0026sup2;⁺ and its status in different solvent were noted as depicted in the below Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab1\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eSolvent Screening and Solubility Profile of carrageenan.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eS. No.\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSolvents\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eObservation\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthyl Acetate*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eEthanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDichloromethane (DCM)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eN, N, dimethyl Formamide (DMF) *\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and partially soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAcetone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel \u0026amp; Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eToluene*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAcetone\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eDimethyl sulfoxide (DMSO)*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eMethanol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAcetonitrile*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel and Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater (Hot)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGel formation\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003e* Temperature was also enhanced till boiling point and similar observation was observed.\u003c/p\u003e\n \u003cp\u003eThe solubility of the Ca\u003csup\u003e+\u0026thinsp;2\u003c/sup\u003e Carrageenan was tested using polar protic (Acetone, Alcohols), aprotic (DMF), high boiling polar solvent (DMSO), and non-polar (toluene) solvents; however, none of the solvents were successful in producing the carrageenan solution phase, Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e (Entry 1\u0026ndash;10), even after raising the temperature to the boiling point of the corresponding solvent. It was anticipated that the polymer would remain in solution phase following dissociation, which was only made possible in water due to the solvent\u0026apos;s high dissociation capacity. After screening with organic solvents, water (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e entry 11) was utilized as an aqueous solvent. Surprisingly, it wasn\u0026apos;t solvable, and the outcomes from entries 1 through 11 were comparable as before.\u003c/p\u003e\n \u003cp\u003eThe temperature of the carrageenan solution in water was gradually raised until it took the form of a gel; however, in the case of K\u003csup\u003e+\u003c/sup\u003e, the gel form was with water at a normal temperature and the solution phase was with a high temperature; the opposite result with Ca\u003csup\u003e2+\u003c/sup\u003e provided a reason to further optimize the temperature effect to see how the behaviour changed as the water temperature changed.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable :2\u003c/strong\u003e Effect of temperature in the water and carrageenan system:\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Taba\" border=\"1\"\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIce-Water\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNormal Water\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHot water\u003c/p\u003e\n \u003cp\u003e48-80\u003csup\u003eo\u003c/sup\u003eC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHigh Temperature 80\u003csup\u003eo\u003c/sup\u003eC\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel, Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eNot Forming Gel, Insoluble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eGel formation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eViscous solution, soluble\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eThe temperature-based analysis shows the pattern of Gel-sol property after the heating, however in case of K\u003csup\u003e+\u003c/sup\u003e the sol-Gel conversion was taking place with respect to the heating. The further investigation of the ion- effect was caried out, by keeping in the mind the Ca\u003csup\u003e2+\u003c/sup\u003eis smaller (2.4 \u0026Aring;) than the potassium (2.7\u0026Aring;) and the computational based study regarding the distance between sulphate ion and K\u003csup\u003e+\u003c/sup\u003e and Ca\u003csup\u003e2+\u003c/sup\u003e as depicted in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e.\u003c/p\u003e\n \u003cp\u003eThe distance between Ca\u003csup\u003e2+\u003c/sup\u003e and carrageenan was less than the K\u003csup\u003e+\u003c/sup\u003e, it was also clue regarding the more attraction or more bond dissociation energy with the Ca\u003csup\u003e2+\u003c/sup\u003eion. It was anticipated that the cold/normal water does not converts in to the ionic form, however the high temperature when matches with the dissociation energy gets in to the dissociation for and forms gel with stable \u003csup\u003e1\u003c/sup\u003eC\u003csub\u003e4\u003c/sub\u003e conformers for the formation of gel. It was also interesting to see that heating more than 80\u003csup\u003eo\u003c/sup\u003eCwas converting Gel to solution, because of its conformational change from \u003csup\u003e4\u003c/sup\u003eC\u003csub\u003e1\u003c/sub\u003e, A higher energy state for solution as it is related with the reported study regarding the role of conformational changes and its effect on phase, as depicted in the below Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e. Moreover, the network with H\u003csub\u003e2\u003c/sub\u003eO which was observe in the IR and Mass also suggest the close linkage of the network in undissociated form, which does not get ionized in the normal temperature so the higher temperature is require to form the gel in with stable \u003csup\u003e1\u003c/sup\u003eC\u003csub\u003e4\u003c/sub\u003e conformer as depicted in Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e.\u003c/p\u003e\n \u003cp\u003eHowever, in the case of K\u003csup\u003e+\u003c/sup\u003e the bond attraction was less so it dissociated in normal water and forms gel, and in case of the hot water it forms solution because of the conformational change to \u003csup\u003e4\u003c/sup\u003eC\u003csup\u003e1\u003c/sup\u003e as solution phase. Figure\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e\u003c/p\u003e\n \u003cp\u003eThe dissociation theory for Ca\u003csup\u003e2+\u003c/sup\u003e was further analyse through the ionic content in the solution with respect to the temperature. The carrageenan and water solution were reacted with the NaOH solution in each temperature and the pattern of the reactivity was found different as per given Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab2\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eReaction of free calcium ion with NaOH in different temperature.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eIce-Water\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNormal Water\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHot water\u003c/p\u003e\n \u003cp\u003e48-80\u003csup\u003eo\u003c/sup\u003eC\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eHigh Temperature 80 \u003csup\u003eo\u003c/sup\u003eC\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter the addition of NaOH, the solution turns viscous.\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter the addition of NaOH, the solution turns more viscous\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter the addition of NaOH solution, it does not give any change\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAfter the addition of NaOH solution, it does not give any change\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eVery light Precipitate forms after 15\u0026ndash;20 mints with less yield\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eLight precipitate forms in 15 minutes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePrecipitate in few minutes with high yield\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePrecipitate in few minutes with high yield\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eThe reactivity pattern with NaOH clearly represent the availability of free calcium ion present in the solution, it also confirms the dissociation of the Ca\u003csup\u003e2+\u003c/sup\u003e is also increasing with high temperature and justifying our plausible mechanism regarding the undissociated form in cold water of Ca-linked carrageenan. The ionic content in hot water was also confirm by the conductivity as the io gets dissociated the conductivity increase and the similar result obtained in the test, and the gelling property correlation with ionized and unionized form was confirm through the experiment. Conductivity based on the principle by measuring the electric current flows in electrons present in the solution means if the solution is higher in ion concentration shows reading towards higher, which is observed in our result, as ca ion are in dissociated form in hot water so showing conductivity more than cold and normal water. However. The conductivity of the K\u003csup\u003e+\u003c/sup\u003e carrageenan remains same in all the condition, so the dissociation was same in the all cases, while with the Ca\u003csup\u003e2+\u003c/sup\u003eit increases with the temperature.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab3\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003econductivity of the water carrageenan solution in different temperatures.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCa CG with water at different temperature\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eReadings\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eHot water (50-100\u003csup\u003e0\u003c/sup\u003eC)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.59\u0026ndash;0.64\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWater (RT)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCold water (8 \u003csup\u003e0\u003c/sup\u003eC) Normal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eThe investigation into the Ca\u003csup\u003e2+\u003c/sup\u003e linked carrageenan and its temperature-dependent role is supported by a number of chemical and experimental evidence, and it is suggested that because the Ca\u003csup\u003e2+\u003c/sup\u003e linked carrageenan has a higher dissociation energy than the K\u003csup\u003e+\u003c/sup\u003e linked carrageenan, it forms gel in hot water and is insoluble in cold water, supporting the role of cations and conformational changes.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\n \u003ch2\u003eViscosity:\u003c/h2\u003e\n \u003cp\u003eThe viscosity of the K\u0026thinsp;+\u0026thinsp;linked gel was found as 649 centipoise and the gel with Ca\u003csup\u003e2+\u003c/sup\u003e has 728, (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e) which also justifies the higher cross linking with helical-to-helical linkage to form more stiff and stable gel. The intermolecular interactions (Fig.\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e9\u003c/span\u003e) were also observed by IR through the OH deformation peak, so the more interactions get more stiff gel.\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\n \u003ctable id=\"Tab4\" border=\"1\"\u003e\n \u003ccaption\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eViscosity results of nano and simple gel\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eType of Gel\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eViscosity\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eK\u003csup\u003e+\u003c/sup\u003eCarrageenan gel\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e649\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eCa\u003csup\u003e+\u003c/sup\u003eCarrageenan gel\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e728\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eIota carrageenan plays important role either in food industry or for medical purposes due to their numerous applications. The effect of cations plays crucial role in case of carrageenan or we can say changes in their physiochemical behavior, that synergizes its effect and application. Also, the type of cation bivalent, monovalent is also responsible for it, affected the gelling property of carrageenan, with calcium here we find not forming gel in normal water while comparing to potassium because of the requirement of high energy for dissociation. Our study finds out the entire parameter proofing the higher energy required for Ca\u003csup\u003e2+\u003c/sup\u003eto dissociate and so forming gel in hot water.\u003c/p\u003e"},{"header":"DECLARATIONS","content":"\u003cp\u003e\u003cstrong\u003eAcknowledge:\u0026nbsp;\u003c/strong\u003eAll authors express their gratitude to Graphic Era Hill University for providing the initial funding to start the project work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Declaration\u003c/strong\u003e: Not any\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest-\u003c/strong\u003e Not any\u003c/p\u003e"},{"header":"REFERENCES","content":"\u003col\u003e\n \u003cli\u003ePurohit P, Bhatt A, Mittal RK, Abdellattif MH, Farghaly TA. Polymer Grafting and its chemical reactions. Frontiers in Bioengineering and Biotechnology. 2023 Jan 11;10:1044927.\u003c/li\u003e\n \u003cli\u003eEvageliou VI, Ryan PM, Morris ER. Effect of monovalent cations on calcium-induced assemblies of kappa carrageenan. Food Hydrocolloids. 2019 Jan 1;86:141-5.\u003c/li\u003e\n \u003cli\u003eMorris ER, Rees DA, Robinson G. Cation-specific aggregation of carrageenan helices: domain model of polymer gel structure. Journal of molecular biology. 1980 Apr 5;138(2):349-62.\u003c/li\u003e\n \u003cli\u003eJanaswamy S, Chandrasekaran R. Heterogeneity in iota-carrageenan molecular structure: insights for polymorph II\u0026rarr; III transition in the presence of calcium ions. Carbohydrate Research. 2008 Feb 4;343(2):364-73.\u003c/li\u003e\n \u003cli\u003eMorris ER, Rees DA, Robinson G. Cation-specific aggregation of carrageenan helices: domain model of polymer gel structure. Journal of molecular biology. 1980 Apr 5;138(2):349-62.\u003c/li\u003e\n \u003cli\u003evan de Velde F, Rollema HS, Grinberg NV, Burova TV, Grinberg VY, Tromp RH. Coil\u0026ndash;helix transition of \u0026iota;‐carrageenan as a function of chain regularity. Biopolymers: Original Research on Biomolecules. 2002 Nov 15;65(4):299-312.\u003c/li\u003e\n \u003cli\u003eRobal M, Brenner T, Matsukawa S, Ogawa H, Truus K, Rudolph B, Tuvikene R. Monocationic salts of carrageenans: Preparation and physico-chemical properties. Food Hydrocolloids. 2017 Feb 1;63:656-67.\u003c/li\u003e\n \u003cli\u003eMacArtain P, Jacquier JC, Dawson KA. Physical characteristics of calcium induced \u0026kappa;-carrageenan networks. Carbohydrate Polymers. 2003 Sep 1;53(4):395-400.\u003c/li\u003e\n \u003cli\u003eVan de Velde F, Knutsen SH, Usov AI, Rollema HS, Cerezo AS. 1H and 13C high resolution NMR spectroscopy of carrageenans: application in research and industry. Trends in Food Science \u0026amp; Technology. 2002 Mar 1;13(3):73-92.\u003c/li\u003e\n \u003cli\u003eLeal D, Matsuhiro B, Rossi M, Caruso F. FT-IR spectra of alginic acid block fractions in three species of brown seaweeds. Carbohydrate research. 2008 Feb 4;343(2):308-16.\u003c/li\u003e\n \u003cli\u003eBongaerts K, Paoletti S, Denef B, Vanneste K, Cuppo F, Reynaers H. Light scattering investigation of \u0026iota;-carrageenan aqueous solutions. Concentration dependence of association. Macromolecules. 2000 Nov 14;33(23):8709-19.\u003c/li\u003e\n \u003cli\u003eJanaswamy S, Chandrasekaran R. Effect of calcium ions on the organization of iota-carrageenan helices: an X-ray investigation. Carbohydrate Research. 2002 Mar 15;337(6):523-35.\u003c/li\u003e\n \u003cli\u003eFontes-Candia C, Str\u0026ouml;m A, Lopez-Sanchez P, L\u0026oacute;pez-Rubio A, Mart\u0026iacute;nez-Sanz M. Rheological and structural characterization of carrageenan emulsion gels. Algal Research. 2020 May 1;47:101873.\u003c/li\u003e\n \u003cli\u003eNickerson MT, Darvesh R, Paulson AT. Formation of Calcium‐Mediated Junction Zones at the Onset of the Sol‐Gel Transition of Commercial \u0026kappa;‐Carrageenan Solutions. Journal of food science. 2010 Apr;75(3):E153-6.\u003c/li\u003e\n \u003cli\u003eTarı \u0026Ouml;, Pekcan \u0026Ouml;. Comparison of cation effects on phase transitions of kappa and iota carrageenan. e-Polymers. 2010 Dec 1;10(1):083.\u003c/li\u003e\n \u003cli\u003eViebke C, Piculell L, Nilsson S. On the mechanism of gelation of helix-forming biopolymers. Macromolecules. 1994 Jul;27(15):4160-6.\u003c/li\u003e\n \u003cli\u003eNeamtu B, Barbu A, Negrea MO, Berghea-Neamțu CȘ, Popescu D, Zăhan M, Mireșan V. Carrageenan-based compounds as wound healing materials. International Journal of Molecular Sciences. 2022 Aug 14;23(16):9117.\u003c/li\u003e\n \u003cli\u003eMontero P, P\u0026eacute;rez-Mateos M. Effects of Na+, K+ and Ca2+ on gels formed from fish mince containing a carrageenan or alginate. Food Hydrocolloids. 2002 Jul 1;16(4):375-85.\u003c/li\u003e\n \u003cli\u003eBui VT, Nguyen BT, Nicolai T, Renou F. Mixed iota and kappa carrageenan gels in the presence of both calcium and potassium ions. Carbohydrate polymers. 2019 Nov 1;223:115107.\u003c/li\u003e\n \u003cli\u003eSeki T, Chiang KY, Yu CC, Yu X, Okuno M, Hunger J, Nagata Y, Bonn M. The bending mode of water: A powerful probe for hydrogen bond structure of aqueous systems. The journal of physical chemistry letters. 2020 Sep 15;11(19):8459-69.\u003c/li\u003e\n \u003cli\u003eGhani NA, Othaman R, Ahmad A, Anuar FH, Hassan NH. Impact of purification on iota carrageenan as solid polymer electrolyte. Arabian journal of chemistry. 2019 Mar 1;12(3):370-6.\u003c/li\u003e\n \u003cli\u003eRajasulochana N, Gunasekaran S. Analysis on the seasonal variations in carrageenans of Hypnea flagelliformis and Sarconema filiforme by FTIR spectroscopy. Asian Journal of Chemistry. 2009 Jul 1;21(6):4547.\u003c/li\u003e\n \u003cli\u003eJumaah FN, Mobarak NN, Ahmad A, Ghani MA, Rahman MY. Derivative of iota-carrageenan as solid polymer electrolyte. Ionics. 2015 May;21:1311-20.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Graphic Era Hill University","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Carrageenan, Polysaccharide, conformational analysis, cation, gelation behavior","lastPublishedDoi":"10.21203/rs.3.rs-3421472/v2","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3421472/v2","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eCarrageenan is a natural polymer extracted from the Red-Seaweeds that is important for the food and pharmaceutical industries. It is crucial to comprehend how the ion exchange affects the rheological properties of Iota carrageenan polymer, particular potential for gelation, which has several applications in the food and pharmaceutical industries. The current study focuses on analyzing the rheological changes brought on by the exchange of potassium ions with calcium ions in Iota carrageenan. Due to their distinct associations with the anion component, monovalent and divalent cations both have a varied impact on the gelation property of carrageenan.\u003c/p\u003e","manuscriptTitle":"Investigation of the rheological alterations in Iota carrageenan through the exchange of potassium with calcium ions.","msid":"","msnumber":"","nonDraftVersions":[{"code":2,"date":"2024-01-03 18:45:35","doi":"10.21203/rs.3.rs-3421472/v2","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}},{"code":1,"date":"2023-10-10 18:40:45","doi":"10.21203/rs.3.rs-3421472/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"525a6b51-1064-47a0-9310-2fcc58e433b4","owner":[],"postedDate":"January 3rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":25292806,"name":"Polymer Science"}],"tags":[],"updatedAt":"2023-10-10T18:40:46+00:00","versionOfRecord":[],"versionCreatedAt":"2024-01-03 18:45:35","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v2","identity":"rs-3421472","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3421472","identity":"rs-3421472","version":["v2"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}