A Study On Encapsulating Biology Essay

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Abstract

Encapsulating compounds with sodium alginate and make granules of different compounds such as glucose, vitaminB12 and aspirin. Alginates and alginic acid have the gel forming property so that they have been most popularly used in many of food industry, pharmaceutical industries, biotechnology and in medicine. The alginate which has gel forming capacity has advantage low toxicity, biocompatible non-immunogenic and biodegradable biopolymer. Molecules which are low molecular weight diffuse more effectively from gel beads. For all the above mention compounds measure the diffusion in water and 0.9%saline at room temperature and at 370C for time intervals of 0,5,10,15,30,60,120,180 and 240. Measure the release with pre-gelatinised starch for the same above mention time intervals.

Introduction

Alginate is described as the linear polysaccharide of (1 -+ 4) linked a-L-guluronate (G) and p-D mannuronate (M) residues which are set in the form of irregular, block wise outline along with linear chain. The block wise pattern consist of three types of polymer segments mainly first one contains D-mannuronic acid units, the second consist of L-guluronic acid units and finally last is alternating D-mannuronic acid and L-guluronic acid residues (Velings and Mestdagh 1994). Alginate is characterized as biopolymer and polyelectrolyte and has properties of biodegradable, non-toxic, biocompatible and non-immunogenic (Yang et al.2007). The physical properties of alginate depend upon source of origin of seaweed, extracting alginate from a part of algae and time of harvesting in a season. The analysis study is not fully done or unknown because of its higher molecular weight and its complex nature. Hence due to growing use and application of alginate in the fields of biomedical and pharmaceutical, it has been important that to use sensitive methods for testing quantity of alginate present in products for quality control processes (Oztekin et.al, 2006).

Sodium alginate molecular structure

Alginate is used as most preferred as biomaterial because of its micro structural qualities such as biocompatible unbranched binary copolymers those are used in fields such as tissue engineering, controlled release, drug delivery, and cell immobilization, immobilization of micro organisms and in food applications. The most important practical application of alginate with surfactants interaction in aqueous solutions was studied (Yang et.al 2007). The physicochemical properties of mixture of alginate and surfactant have widely used in variety industrial fields such as biotechnology, medicine and food industry because of its gel forming characteristic (Oztekin et.al, 2006).

Alginate beads which are insoluble in water are formed by adding dropwise alginate solution into a solution containing divalent metal ions. The formed beads are thermo irreversible compounds and doesn't dissolve in water but they can able to dissolve in chemicals agents like EDTA which as cation squestring. The model developed by Grant et d4 is called as egg box which usually tells us that divalent metal ions bounds in interchain cavities in fundamentally polygulurinate sequences that will give a structure as rod like cross linked complex (Velings and Mestdagh 1994).

Encapsulation

Encapsulation is defined as it provides as shield, concentrate and protects molecules by providing microenvironment and forming layer with biopolymer and the facts for appearance of encapsulation was unclear.

According to the patent described in US 3645911 in Feb 26 1972 tells that solid layer of polymeric substance forms a microcapsules which contain hydrophilic substances inside capsule are formed by three different ways. They are defined as dispersing or emulsifying hydrophilic compounds into the solution of polymeric materials, then the solvent for a particular substance are being water- immiscible and having boiling point lower than 1000C.

Emulsifying the emulsion or dispersion obtained in an aqueous solution of hydrophilic colloid and removing solvent from polymeric material from the system so formed by evaporation.

The important aqueous phase which is used for filling inside capsule is defined as first aqueous phase and the biopolymer which is used for creating particular environment for which it is surrounded by the first aqueous phase in which the droplets used such phase is called as second aqueous phase.

It has been proved that first phase of aqueous dispersion of substance in the aqueous solution of hydrophilic colloid make the substance to move into first phase of aqueous solution to second phase occurs because of this transfer given concentration of encapsulation cannot be done. Because of this reason it has been found that encapsulation of aqueous solution without removal or washing out dissolved substance which is used to fill inside capsule during encapsulation process (Pat. No 3645911, 1972).

According type of compounds used for encapsulation its morphology physical and chemical properties have the significance effect on the type of methods which should be used for encapsulation. The most important techniques used for encapsulation are as follows they are solvent evaporation, interfacial polymerization and matrix entrapment.

Interfacial polymerization is characterized as polymerization at solvent interface and water of two different monomers. Monomers from two different monomers are dissolved in water phase and another is dispersed in organic solvent. This method encapsulates a liquid core which is formed as core capsule product. One of the cons of this method is denaturation of sensitive biomolecules is done with the help of aggressive chemicals and organic liquids.

Solvent evaporation method is characterized as physical method where there is no need of chemicals hence no chance chemical reactions. But have the same disadvantage as interfacial polymerization method.

Both methods contain about 30-75% of core product as encapsulated substance.

Matrix entrapment method is defined as biomolecules that are entrapped into a meshwork of a gel where the substances are fixed into a gel which have the pore size less than size of substance used. Substance that are used for this method are particles, macromolecules, organelles, cells and microbes. The main pros of this method are gel used in this method has low toxicity for entrapment of living organisims. But compounds used which are low molecular weight have chance of diffusion out of gel hence it can't be prevented. Entire volume of capsule is enclosed with gel matrix as significant part.

Other than the above discussed encapsulation methods, some of other methods depended on extruding of plasticized substances, transport of compounds into pre produced capsules or combination are used which were mainly viewed on application in biomedical and pharmaceuticals.

Hence in life sciences and in industrial application encapsulation or immobilization of biomaterial has raised higher interest. During earlier period biomaterial were encapsulated as a solution. It is described in general as biomaterials for encapsulation in solid form and immobilization strategy depended on biomaterial encapsulated. Water soluble biomaterials which are highly soluble such as enzyme glucose oxidase has preferred as biomaterial substance. (Trau and Renneberg 2003)

Glucose

Diffusion is often considered to be a rate-limiting step in many processes. It is thus important to determine the diffusion coefficient in order to estimate the total rate of mass transfer. A low mass transfer rate will usually be a drawback, except in some areas, such as slow and controlled drug release and in packaging technology (Anderson et.al 1997)

Methods and materials

Materials and chemicals:

Sodium alginate, 0.5M CaCl2, 0.9% saline, glucose kit, redistilled water, spectrophotometer, glucose, vitaminB12 (cyanocobalamin) , aspirin(acetyl acetylsalicylic acid).

Preparation of blank sodium alginate beads: To 100ml of redistilled water add about 3g of sodium alginate and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of sodium alginate with 3% glucose beads: To 100ml of redistilled water add about 3g of sodium alginate and 0.3g of glucose and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of sodium alginate with 3% vitaminB12 beads: To 100ml of redistilled water add about 3g of sodium alginate and 0.3g of vitamin B12 and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of sodium alginate with 3% aspirin beads: To 100ml of redistilled water add about 3g of sodium alginate and 0.3g of aspirin and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of blank sodium alginate beads with 3% pre gelatinized starch (ps): To 100ml of redistilled water add about 2.7g of sodium alginate with 0.3g of starch and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of 3% glucose sodium alginate beads with 3% pre gelatinized starch (ps): To 100ml of redistilled water add about 2.7g of sodium alginate with 0.3g of starch and to it add 0.3g of glucose and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of 3%vitaminB12 sodium alginate beads with 3% pre gelatinized starch (ps): To 100ml of redistilled water add about 2.7g of sodium alginate with 0.3g of starch and to it add 0.3g of vitaminB12 and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Preparation of 3% aspirin sodium alginate beads with 3% pre gelatinized starch (ps): To 100ml of redistilled water add about 2.7g of sodium alginate with 0.3g of starch and to it add 0.3g of aspirin and mix them for 60min by using magnetic stirrer until both mixes completely. By using syringe drop sodium alginate solution into 0.5M CaCl2solution.The CaCl2solution was stirred at uniform speed using magnetic stirrer while dropping alginate. Thus they formed granules were filtered by using filter paper into petri plates and allow them to dry under 370C oven for overnight.

Weigh 1g of each the sample granules and drop them into redistilled water and 0.9% saline at room temperature and 370C. By using spectrophotometer measure absorbance at 340nm for glucose, 360nm for vitaminB12 and 285nm for aspirin at time intervals of 0, 5, 10, 15, 30, 60, 120, 180 and 240minutes.

Glucose cannot be measured directly with spectrophotometer because of its low molecular weight rays were not able to detect so we used glucose kit. It consists of 3 solutions which are used according to procedure suggested along with kit.

Results:

Concentration of glucose in water and 0.9% saline at room temperature and 370C at 340nm.

Time

Conc.of glucose in water at 37

Conc. of glucose in saline at 37

Conc. glucose in water at room temp

Conc. glucose in saline at room temp

0

0.001727

0.002015

0.009214

0.002591

5

0.002593

0.019

0.056427

0.057289

10

0.06246

0.029075

0.07341

0.07197

15

0.06966

0.054122

0.075425

0.080607

30

0.07628

0.059876

0.082047

0.08073

60

0.07889

0.076577

0.084062

0.079168

120

0.0794

0.077728

0.084062

0.080895

180

0.08003

0.079737

0.08435

0.081471

240

0.08003

0.079737

0.084062

0.080893

Concentration of glucose in alginate plus 3% pre gelatinized starch at 370C

Time

conc. of glucose in water

conc. of glucose in saline

0

0

0

5

0.000863

0.040016

10

0.032813

0.061031

15

0.071395

0.06391

30

0.075713

0.068804

60

0.076001

0.069956

120

0.076289

0.069956

180

0.076577

0.070243

240

0.077153

0.070531

Absorbance of vitaminB12 in water and 0.9%saline at 360nm at room temperature and 370C.

Time in minutes

Absorbance in water at 370C

Absorbance in0.9% saline at 370C

Absorbance in water at room temp

Absorbance in 0.9% saline at room temp

0

0

0

0.002

0.001

5

0.242

0.239

0.183

0.164

10

0.329

0.345

0.327

0.321

15

0.352

0.37

0.383

0.37

30

0.389

0.407

0.44

0.429

60

0.397

0.425

0.473

0.465

120

0.391

0.419

0.481

0.478

180

0.373

0.401

0.472

0.469

240

0.371

0.383

0.457

0.457

Absorbance of aspirin with alginate and alginate plus 3%pre gelatinised starch in saline at room temperature and 370C at 285nm

Time

Absorbance at room temp

Absorbance at 37

Absorbance in at room temp ps

Absorbance at 37 ps

0

0.078

0.011

0

0.006

5

0.403

0.425

0.464

0.475

10

0.586

0.604

0.58

0.688

15

0.665

0.631

0.675

0.747

30

0.691

0.681

0.706

0.805

60

0.796

0.661

0.758

0.802

120

0.751

0.66

0.723

0.795

180

0.693

0.658

0.729

0.792

240

0.696

0.63

0.693

0.756

Conclusion:

The release of glucose is faster when compared with vitaminB12 and aspirin.

Reference :

1. Nicolas M. Velings & Michkle M. Mestdagh. Physico-Chemical Properties of Alginate Gel Beads. 1994, Source: Polymer Gels and Networks, Vol. 3, Issue 3, Pages 311-330.

2. Jisheng Yang, Jianyu Zhao and Yun Fang. Calorimetric studies of the interaction between sodium alginate and sodium dodecyl sulfate in dilute solutions at different pH values, 2007; Source: Carbohydrate Research,Volume 343, Issue 4, 17 March 2008, Pages 719-725

3. Nevin O¨ ztekin, Selda Bas¸kan, F. Bedia Erim. Determination of alginate copolymer in pharmaceutical formulations by micellar electrokinetic chromatography, 2006. Source: Journal of Chromatography B Volume 850, Issues 1-2, Pages 488-492

4. Jan Frans van Besauw, Brasschaat and Daniel Alois Claeys. US patent 3645911. Methods for encapsulating aqueous or hydrophilic material. 1972

5. Dieter Trau and Reinhard Renneberg. Encapsulation of glucose oxidase microparticles within a nanoscale layer-by-layer film: immobilization and biosensor applications 2003.Volume 18, issue 12, Pages 1491-1499.

6. Matz Andersson, Anders Axelsson , Guido Zacchi. Diffusion of glucose and insulin in a swelling N-isopropylacrylamide gel, 1997.Source :International Journal of Pharmaceutics

 

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