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Slow Release Characteristics of β-Glucan

Liqing Lai, Vanja Perčulija, Jia Wu, Lan Zhao
Journal of Food and Nutrition Research. 2020, 8(2), 102-109. DOI: 10.12691/jfnr-8-2-5
Received December 25, 2019; Revised February 04, 2020; Accepted February 21, 2020

Abstract

β-glucan freeze-thaw gel was used as a carrier to release curcumin. The results showed that the freeze-dried gel had the ideal sustained release following 5% β-glucan solution by adding 1% β-glucan quality curcumin, repeated freezing and thawing 8 times. The Peppas equation can significantly describe the release mechanism of curcumin in the β-glucan gel. This study further demonstrated that the release of curcumin in the oat β-glucan gel is the Fick diffusion mechanism and coated barley β-glucan is a non-Fick spread.

1. Introduction

Curcumin is a plant polyphenolic substance, and it is a relatively small pigment with a diketone structure. It was initially extracted from ancient chinese medicine and has anti-oxidation, liver protection, and anti-inflammatory effects. It has anti-cancer, hypolipidemic, anti-thrombotic and anti-peroxide effects, and has no side effects on the body. It is considered to be one of the ideal natural anti-cancer compounds 1, 2, 3, 4, 5. However, due to its poor water solubility, it has shortcomings such as low absorption in the body, rapid metabolism, and low bioavailability, which limits its application. The sustained release dosage form has the characteristics of prolonging the action time of the drug, increasing the therapeutic index, reducing the dosage, reducing the toxicity, facilitating the administration of the drug and increasing the adaptability of the patient, so it has broad applications, prospects and development trends 6, 7, 8.

β-glucan can form a gel under certain conditions, but the microstructure of the β-glucan molecule, the influence of the high-order structure on the gel formation mechanism, and the gel properties and applications need further research. It provides a theoretical basis for the practical application of β-glucan in the food industry, provides a theoretical basis for the finishing of oats and barley, and contributes to the development of grain science 9, 10.

With the application of polymer materials in medicine and further research on the mechanism of action of drugs, sustained-release drug preparations have begun to be widely used in clinical practice. The use of gel sustained release agents has many advantages over conventional pharmaceutical preparations 11, 12. The treatment time for humans is relatively long such as the frequency of administration of drugs is relatively small, the stimulation to the gastrointestinal tract is shortand has no toxic side effects. In addition, the fluctuation of the peak period is short and the release of the oral drug in the stomach and the small intestine can be avoided. Likewise, the stimulation of the gastrointestinal mucosa caused by repeated use of the drug and the reduction of systemic side effects can be avoided. Similarly, the effectiveness and safety of the drug are greatly improved. The emergence of controlled release and sustained release drugs to a certain extent meet the needs of patients with chronic seizures. They not only can avoid side effects, but are also easy to take especially for patients with severe medications such as children and the elderly. This project intends to use β-glucan to encapsulate curcumin to study the sustained release process of curcumin in β-glucan gel, and use β-glucan as a completely natural carrier for β-glucan 13, 14, 15, 16, 17, 18.

2. Materials and Methods

2.1. Preparation of Curcumin Solution

A certain mass fraction of curcumin solution was prepared by using 90% aqueous ethanol solution, Tween-80 ethanol solution (Tween 80 mass fraction 10%) and Tween-80 MCT solution (Tween-80 mass fraction 10%). Unless specified in all the following experiments, curcumin was prepared with Tween-80 ethanol solution (Tween-80 mass fraction 10%), and then added to the β-glucan solution, the Tween-80 mass fraction in the final system 0.5%, ethanol volume fraction was 4.5%.

2.2. Preparation of Sustained Release Gel
2.2.1. Different Curcumin Dissolution Methods

Prepared OG0 and BG0 solutions with a mass fraction of 4%. Curcumin was dissolved in a mixed liquid of 90% aqueous ethanol solution, Tween-80 aqueous solution (mass fraction 10%) and Tween-80 MCT (Tween-80 mass fraction 10%). The β-glucan solution was magnetically stirred at 85 °C for 2.5 h, then added with 1% β-glucan containing curcumin, dissolved for 0.5 h, freeze-thawed 8 times, and then freeze-dried.


2.2.2. Different Dosage

Prepared 4% OG0 and BG0 solutions, magnetically stirred the β-glucan solution at 85 °C for 2.5 h, then added 0.5%, 1.0% and 1.5% (w/w) β-glucan to supplement turmeric, re-dissolved for 0.5 h, freeze-thaw 8 times, and freeze-dried.


2.2.3. Different Molecular Weight β-Glucans

Prepared OG0, OG30, OG60, OG90 and BG0, BG30, BG60, BG90 solutions with a mass fraction of 4%. The β-glucan solution was magnetically stirred at 85 °C for 2.5 h, then added 1% β-glucan contaning curcumin, redissolved for 0.5 h, freeze-thawed 8 times and freeze-dried.


2.2.4. Different Freeze-Thaw Cycles

Prepared 4% OG0, BG0 solution. The β-glucan solution was magnetically stirred at 85 °C for 2.5 h, then added 1% β-glucan to supplement curcumin, then dissolved for 0.5 h, freeze-thawed 4 times and freeze dried after 6, 8, and 10 times.


2.2.5. Different Mass Fraction of β-Glucan

Prepared OG0 and BG0 solutions with mass fractions of 3%, 4%, and5%. The β-glucan solution was magnetically stirred at 85 °C for 2.5 h, then added 1% β-glucan to supplement curcumin, then dissolved 0.5 h, freeze-thawed 8 times and freeze-dried.

2.3. Determination of the Sustained Release of Curcumin
2.3.1. Simulated Gastric Juice Preparation

The 0.1 g/L hydrochloric acid solution (containing 0.5% Tween-80) was used as a simulated gastric juice to study the release of curcumin from the β-glucan gel. According to relevant literature studies, curcumin is relatively stable in an acidic toa neutral environment. Under alkaline conditions, the hydroxyl group on the phenyl ring of curcumin is in the form of oxygen anion, which has strong electron donating ability, resulting in enhanced electrophilic reactivity of the carbon chain, so it is unstable under alkaline conditions, so it is mainly simulated sustained release


2.3.2. Curcumin Detection and Wavelength Selection

Weighed 0.01 g of curcumin sample, diluted to 100 mL with absolute ethanol, took 5 mL and diluted to 50 mL with simulated gastric juice to obtain 50 μg/mL curcumin solution. Then took 5 mL and diluted to 10 mL with simulated gastric juice. Baseline was calibrated with simulated gastric fluid, and UV scanning was carried out in the 300-600 nm wavelength range.


2.3.3. Drawing of Curcumin Standard Curve

The vacuum was weighed to a constant weight of curcumin to a 100 mL volumetric flask, dissolved in absolute ethanol and diluted to volume. Then accurately transferred 5 mL to 50 mL brown volumetric flask, and diluted to volume with simulated gastric juice to prepare 10.14 μg/mL of standard curcumin solution. Next, weighed 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0 and 5.0 mL in a 10 mL volumetric flask, diluted with simulated gastric fluid, simulated gastric juice as a blank, and used UV spectrophotometer. The absorbance was measured at the maximum absorption wavelength. Taking the mass fraction as the abscissa, the corresponding absorbance was plotted on the ordinate and regression analysis was performed to obtain the standard curve regression equation of curcumin in simulated gastric juice.


2.3.4. Determination of Curcumin Release Rate

The simulated gastric juice was used as the release medium. The curcumin-containing β-glucan sustained-release gel was placed in a simulated gastric environment (0.1 mol/L hydrochloric acid, containing 0.5% Tween-80), oscillated at 37 ° C (at a constant temperature) 50 r/min), and subjected to a release test. Dissolution medium of 5 mL was taken at a predetermined time, and the absorbance of the solution was measured at a wavelength of 430 nm with an ultraviolet spectrophotometer. Later, an equal volume of gastric juice was added to the release system. The mass fraction of curcumin contained in the solution was calculated according to the standard curve, and the cumulative release was calculated. The cumulative release was plotted on the ordinate and the time was plotted on the abscissa. Accumulation Release Percentage (ARP) was calculated according to formula.

3. Results and Discussion

3.1. Determination of Curcumin Detection Wavelength

The UV scan of curcumin in a simulated gastric juice at 300-600 nm is shown in Figure 1. It can be seen from the figure that curcumin has a maximum absorption peak at 430 nm, baseline at this wavelength is relatively flat, and the interference is very small which is suitable for the requirements of drug testing. Therefore, the detection wavelength of 430 nm as curcumin is determined in the simulated gastric juice which is consistent with the results reported in the literature.

3.2. Drawing of Curcumin Standard Curve

Figure 2 shows the UV absorption standard curve of curcumin in simulated gastric fluid. The linear regression equation is y=1.3443x+0.0026, R2=0.9999. Curcumin has a good linear relationship in the range of 0-0.5 μg/mL mass fraction.

3.3. Slow Release Characteristics Analysis

Unless otherwise specified in the following single factor tests, the preparation conditions of the sustained release gel were as follows: β-glucan samples were OG0 and BG0, β-glucan was added in an amount of 4%, and curcumin was dissolved in Tween-80 ethanol. The solution (Tween-80 mass fraction: 10%) was added to the β-glucan solution, and the amount of curcumin added was 1% of the amount of β-glucan added, and the polysaccharide gel was prepared by repeated freeze-thaw cycles. The Tween-80 mass fraction in the final system is 0.5%, and the ethanol volume fraction is 4.5%.


3.3.1. Effect of Different Curcumin Dissolution Methods

It can be seen from Figure 3 that when the curcumin dissolved in ethanol is added to the system, the release is the fastest and the release rate is the highest, and the ethanol and water can be completely mutually soluble, and the curcumin can be quickly released. The release of curcumin in Tween-80 ethanol is slower, mainly because of Tween-80 forms micelles in aqueous solution. According to the related literature, curcumin may be dispersed in the micelle formed by Tween-80, which has excellent stability. The presence of micelles slows the dissolution and diffusion of curcumin in the release medium. It can be seen from Figure 3 that the release of curcumin in the mixed solution of MCT and Tween-80 is the slowest, and Tween-80 causes the MCT to be dispersed in an aqueous solution to form an emulsified system, which increases the release of curcumin from the MCT to the aqueous phase. This results in a decrease in the rate of curcumin release. However, since MCT is an oily substance, it is not very compatible with the whole system, and the emulsion system formed by Tween-80 and MCT is not conducive to the stability of curcumin. Therefore, it is better to choose the Tween-80 ethanol solution as the solvent of curcumin.


3.3.2. Effect of Dosing

As shown in Figure 4, the release rate of the drug within 2 h of drug release is accelerated with the increase of drug loading. The main reason is that the drug was absorbed on the gel surface. Moreover, the curcumin which is initially released is adsorbed on the surface of the gel, so the release rate is fast, and it is known that the drug loading has a great influence on the initial release rate of the drug. The release profile of curcumin within 2 to 4 hours showed that the drug release rate did not change significantly when the drug loading increased from 0.5% to 1%. When the drug loading was increased to 1.5%, the drug release rate was accelerated. When the drug loading was 0.5% and 1%, the curcumin was mainly distributed in the Tween-80 micelles, and the maximum drug loading of the micelles was not reached. Therefore, the drug release rate was not much different, while the drug loading was 1.5%. At the time, the drug loading exceeds the maximum drug loading of the micelle, and more part of the curcumin is present in the gel structure outside the micelle. When the drug is released, it first appears as the release of the gel surface drug, followed by the diffusion of the curcumin encapsulated in the gel, which is manifested by an increase in the rate of drug release. In the later stage of drug release, the drug loading had no significant effect on the release rate of curcumin 19. Considering that 1.5% of the release of pre-release is too large, 0.5% of the release of the drug is too small, so the choice of 1% of curcumin is appropriate.


3.3.3. Effect of Molecular Weight of β-Glucan

As shown in Figure 5, for oat β-glucan or barley β-glucan alone, the higher the molecular weight, the slower the release of curcumin. It may be because the hydrophilic gel matrix material forms a gel layer due to hydration after contact with water, and curcumin is released by diffusion through the gel layer, and may also be released by gel dissolution.The quality of the gel layer formed depends mainly on the viscosity of the matrix material. The viscosity is large, the gel layer is thick, the diffusion of the drug and the dissolution of the skeleton are slow, and the sustained release effect is slow. The β-glucan gel is a gel-type skeleton, and the viscosity is also increased as the molecular weight is more substantial, which results in a greater hindrance to drug release and a lower release rate. From the theory of polymer structure, the larger the molecular weight, the longer the polymer chain link, the greater the attraction between the links (mainly van der Waals force), the greater the elastic shrinkage energy generated during swelling, and the restriction of hydrophilic condensation. The swelling of the glue. Therefore, the larger the molecular weight of the β-glucan gel, the greater the mechanical strength of the gel, the lower the degree of swelling of the gel matrix, and the slower the release of the drug.


3.3.4. Effect of Molecular Weight of β-Glucan

As shown in Figure 6, within a certain number of freeze-thaw cycles, the sustained release rates of oat β-glucan and barley β-glucan gels slowed down as the number of freezes increased. Mainly because when the number of freeze-thaw cycles increases, the formation of cross-linking structure liquid increases, and the network structure of the gel is more compact which reduces the network gap per unit volume and reduces the size of the pore structure, so turmeric diffusion and release of the hormone will be slower. However, as the number of freeze-thaw cycles increases, the number of times exceeds a certain value, the gel structure is destroyed by ice crystals, the gel mesh is wider and larger, and curcumin is more easily released into the sustained-release medium, further accelerating the release rate. Combined with the previous PCA analysis, it can be seen that when the OG0 and BG0 gel states of the freeze-thaw 8 times have been stabilized, the gel has completely formed. Therefore, the β-glucan gel wrapped with curcumin 8 times of freeze-thaw is selected as a sustained-release preparation


3.3.5. Effect of β-Glucan Mass Fraction

As shown in Figure 7, in general, in the early stage of drug release, since the curcumin adsorbed on the surface of the gel is first released, it is expressed as a diffusion-based release method, so the release is faster. In the later stage of release, curcumin is mainly present in the Tween-80 micelles. As the dextran gel swells and the Tween-80 micelles diffuse from the inside to the outside of the gel, the drug is released, so the release is slower. The higher the mass fraction of oat β-glucan and barley β-glucan, the slower the release of curcumin in the gel formed in the simulated gastric juice may be due to the increase in the mass fraction of β-glucan leading to the gel. The increase in the degree of physical cross-linking results in a denser gel network that hinders the diffusion of Tween-80 micelles containing curcumin and the external curcumin of micelles, It results in a lower rate of drug release. In addition, the sample after lyophilization of the low-mass fraction β-glucan sustained-release gel is loose and partially dispersed in the solution, so that the release of curcumin is accelerated. During the preparation of the gel, it was found that low mass fractions of OG0 and BG0 were difficult to form a solid gel, and the system had a certain solution, not a whole gel. OG0 and BG0 with higher mass fractions are more likely to form a whole gel due to increased cross-linking with each other and have a denser network structure. Curcumin is more difficult to release from the mesh, so the release is slower. The same mass fraction of OG0 is released faster than BG0 because BG0 has a larger molecular weight. Furthermore, the gel structure formed is denser, the degree of cross-linking is higher, and it is difficult to swell, resulting in a slower release of curcumin.

3.4. Slow Release Mechanism Analysis
3.4.1. Slow Release Data Model Fitting

According to the regulations of the State Food and Drug Administration, the development of any sustained-release drugs must be carried out in vitro to determine the sustained release characteristics in vivo. At present, the Chinese Pharmacopoeia's criteria for evaluating the sustained-release model are based on the fact that the correlation coefficient (R) is the largest and the mean square error (MSE) is the best, but in practical applications, the R-value and the model are found in the number of parameters in the middle. When a parameter is added to the model, the R becomes larger, and the model at the moment is not necessarily better. To solve this problem, a widely used information volume criterion, the Akaike Information Criterion (abbreviated as AIC), has been introduced. This criterion is particularly prominent in the selection of statistical models. The AIC minimization method, for a set of data, when the model is fitted to the same extent, with as few parameters as possible 21. The formula for AIC is:

Where RSS is the sum of squared residuals, n is the sample size, and P is the number of independent variables in the regression equation.

According to the samples of 5% OG0 and BG0, the curcumin of 1.0% polysaccharide was added. After 8 freeze-thaw cycles, the cumulative release rate of the gel obtained by freeze-drying in simulated gastric juice was measured by zero-order slow release model. The slow release model, the Higuchi equation, and the Peppas equation were used to fit the sustained release data to establish a sustained release model. The results of the fitting are shown in Table 1 and Table 2.

It can be seen from Table 3 and Table 4 that the data fitting is performed by the Peppas equation, the regression coefficient R of 5% OG0 is 0.95 or more, and the regression coefficient R of 5% BG0 is 0.99 or more. Whether OG0 or BG0, peppas fits the smallest AIC value, so the peppas equation is used to fit the in vitro release data.


3.4.2. Peppas Equation Fitting of Sustained Release Data

There are three main mechanisms for the release of drugs from hydrophilic gel matrix tablets 22: (1) diffusion through the gel layer; (2) dissolution of the gel skeleton; (3) interaction of diffusion and dissolution. Mainly through the diffusion and dissolution methods to achieve the purpose of release. The method gradually dissolves the drug by diffusion of the active ingredient and erosion of the gel matrix layer. When the drug is difficult to dissolve in water, it appears as the erosion of the gel layer. In general, the drug release mechanism can be expressed by the peppas formula. In this paper, the peppas empirical formula was used to fit the release data of β-glucan sustained-release gel in simulated gastric juice, as shown in Table 3 and Table 4.

It can be seen from Table 3 and Table 4 that under the same influencing factors, the release rate is related to the k of the fitting equation, and the faster the release rate, the smaller the k. In contrast, the value of n is consistent with the actual release rate, and the value of n with a fast release rate is also large. According to theory, in cylindrical preparations, when n<0.45, the sustained release mechanism is Fick diffusion; when 0.45 < n < 0.89, it is non-Fick diffusion (drug diffusion and skeleton dissolution together); when n>0.89, it is the framework dissolution mechanism . Therefore, it can be obtained from the table that the n-values of the fitting equations of the in vitro release data obtained by all the factors in the OG are all less than 0.45, and the n values in the BG are mostly between 0.45 and 0.89, and the n values in the lower molecular weight BG are less than 0.45. The release mechanism of OG and BG with lower molecular weight is Fick diffusion, and the release mechanism of BG is non-Fick diffusion, that is, drug diffusion and skeleton dissolution.

4. Conclusions

By single-factor test, 5% β-glucan solution was prepared with OG0 and BG0, and then curcumin dissolved in Tween-80 ethanol solution (Tween-80 mass fraction 10%) was added in an amount of β-glucan 1% of the amount of dextran added. The freeze-thaw method was repeatedly frozen and thawed 8 times, and finally, the dry gel sample was obtained by vacuum freeze-drying. The sample prepared under the conditions had an ideal sustained release effect. Peppas empirical formula Q=ktn can describe the sustained release mechanism. The faster the release rate, the smaller the k value and the larger the value of n. The sustained-release data can be obtained by the Fick diffusion of the oat β-glucan gel encapsulated with curcumin, and the non-Fick diffusion of the cyanine β-glucan cogel containing the curcumin.

Conflict of Interest

The authors declare that there are no conflicts of interest.

Acknowledgements

This work was supported by the Education Department Foundation of Fujian Province of China (JT180081) and FujianProvincial Department and Bureau Project (PP201803). The authors would like to thank AiMi () for providing linguistic assistance.

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Published with license by Science and Education Publishing, Copyright © 2020 Liqing Lai, Vanja Perčulija, Jia Wu and Lan Zhao

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Cite this article:

Normal Style
Liqing Lai, Vanja Perčulija, Jia Wu, Lan Zhao. Slow Release Characteristics of β-Glucan. Journal of Food and Nutrition Research. Vol. 8, No. 2, 2020, pp 102-109. http://pubs.sciepub.com/jfnr/8/2/5
MLA Style
Lai, Liqing, et al. "Slow Release Characteristics of β-Glucan." Journal of Food and Nutrition Research 8.2 (2020): 102-109.
APA Style
Lai, L. , Perčulija, V. , Wu, J. , & Zhao, L. (2020). Slow Release Characteristics of β-Glucan. Journal of Food and Nutrition Research, 8(2), 102-109.
Chicago Style
Lai, Liqing, Vanja Perčulija, Jia Wu, and Lan Zhao. "Slow Release Characteristics of β-Glucan." Journal of Food and Nutrition Research 8, no. 2 (2020): 102-109.
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[1]  Chen F, Liu J, Ye F, et al. Synthesis and characterization of fatty Acid oat β-Glucan ester and its structure–curcumin loading capacity relationship[J]. Journal of Agricultural and Food Chemistry, 2014, 62(50): 56-64.
In article      View Article  PubMed
 
[2]  Jianhua Ming, Jinyun Ye, Yixiang Zhang, Qiyou Xu, Xia Yang, Xianping Shao, Jun Qiang, Pao Xu. Optimal dietary curcumin improved growth performance, and modulated innate immunity, antioxidant capacity and related genes expression of NF-κB and Nrf2 signaling pathways in grass carp ( Ctenopharyngodon idella ) after infection with aeromonas hydrophila[J]. Fish and Shellfish Immunology, 2020, 97.
In article      View Article  PubMed
 
[3]  Rohman A, Sudjadi, Devi, et al. Analysis of curcumin in curcuma longa and curcuma xanthorriza using FTIR spectroscopy and chemometrics[J]. Research Journal of Medicinal Plant, 2015(3): 155-160.
In article      
 
[4]  Hrmova M, Varghese J N, Høj P B, et al. Crystallization and preliminary x-ray analysis ofbeta-glucan exohydrolase isoenzyme exoI from barley (Hordeum vulgare)[J]. Acta Crystallographica, 1998, 54(Pt 4): 687-689.
In article      View Article  PubMed
 
[5]  Anitha A, Maya S, Deepa N, et al. Efficient water soluble o-carboxymethyl chitosannanocarrier for the delivery of curcumin to cancer cells[J]. Carbohydrate Polymers, 2011, 83(2): 452-461.
In article      View Article
 
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