The effect of temperature and chitosan concentration during storage on the growth of chitosan nanoparticle produced by ionic gelation method Wenny Rinda Handani, Wahyudi Budi Sediawan, Ahmad Tawfiequrrahman, Wiratni, and Yuni Kusumastuti
Citation: AIP Conference Proceedings 1840, 080001 (2017); doi: 10.1063/1.4982299 View online: http://dx.doi.org/10.1063/1.4982299 View Table of Contents: http://aip.scitation.org/toc/apc/1840/1 Published by the American Institute of Physics
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THE EFFECT OF TEMPERATURE AND CHITOSAN CONCENTRATION DURING STORAGE ON THE GROWTH OF CHITOSAN NANOPARTICLE PRODUCED BY IONIC GELATION METHOD Wenny Rinda Handani1, Wahyudi Budi Sediawan1, Ahmad Tawfiequrrahman1, Wiratni1,2 and Yuni Kusumastuti 1,2* 1 Chemical Engineering Department, Universitas Gadjah Mada, Yogyakarta Center of Advanced Material and Mineral Processing, Chemical Engineering Department, Universitas Gadjah Mada, Yogyakarta
2
*
Corresponding author:
[email protected]
ABSTRACT The objective of this research was to get the mechanism of nano size chitosan particle growth during storage by observing the effect of temperature and initial concentration of chitosan. The products were analyzed using PSA to have the average of particle radius. Nanochitosan solution was prepared by ionic gelation method. This method is described as an electrostatic interaction between positively charged amine with negatively charged polyanion, such as tripolyphosphate (TPP). Chitosan was dissolved in 1% acetic acid and was stirred for 30 minutes. Tween 80 was added to avoid agglomeration. TPP was prepared by dissolving 0.336 g into distilled water. The nano size chitosan was obtained by mixing TPP and chitosan solution dropwise while stirring for 30 minutes. This step was done at 15qC and ambient temperature (about 30qC) and chitosan concentration 0.2%, 0.4% and 0.6%. The results show that temperature during ionic gelation process (15qC and 30qC) does not affect the initial size of the nanoparticles produced as well as the growth of the nanoparticles during storage. On the other hand, initial chitosan concentration strongly affects initial size of the nanoparticles produced and the growth of the nanoparticles during storage. The concentration of chitosan at 0.2%, 0.4%, 0.6% gave initial size of nanoparticle chitosan of 175.3 nm, 337.9 nm, 643.3 nm respectively. On the other hand, the growth mechanism of chitosan nanoparticle depended on its radius(R). At R500 nm, it is controlled by diffusion in the liquid film around the particles. Keywords : crustacean waste, nano-chitosan, ionic gelation method, TPP, nanoparticle growth model
INTRODUCTION According to the Center of Statistics Data and Information, Department of Marine and Fishery in 2014, it was stated that shrimp and crab export volume reached 90,000 and 4,000 tons respectively. Approximately 80 – 90% of shrimp exported in the form of frozen shrimp without head and shell which is resulting waste with weight reached 25 – 30% of the whole shrimp (Fadli, 2015). On the other side, the waste could be used as raw material for producing chitin, chitosan, and its derivative which is useful and valuable. Another problem of preservative practice fish performed by traditional fishermen is the use of hazardous substances such as formaldehyde. The use of formaldehyde as a food preservative is strictly prohibited by the Regulation of the Minister of Health No. 722/Menkes/Per/IX/1988. Therefore, it is mandatory to find safer ways for maintaining the quality and freshness of the fish. Chitosan has a broad usability in daily life such as biochemistry, medicine, pharmacology, food and nutrition, agriculture, microbiology, waste water treatment, paper industry, membrane or film of textile, cosmetics, and so forth (Illum, 1998). Chitosan is naturally biopolymer which has advantages such as biocompatible, biodegradable, non-
International Seminar on Fundamental and Application of Chemical Engineering 2016 (ISFAChE 2016) AIP Conf. Proc. 1840, 080001-1–080001-7; doi: 10.1063/1.4982299 Published by AIP Publishing. 978-0-7354-1510-2/$30.00
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toxic, and antibacterial (Mardliyati et al., 2012). Physical modification with changing particle size of chitosan to become nanoparticle can be conducted to increase the effectiveness of the good properties of chitosan as anti-microbial (Komariah, 2013). Chitosan has the chemical name of Poly D-glucosamine (β (1-4) 2-amino-2-deoxy-D-glucose) as shown in Figure 1. Chitosan is produced from de-proteinase process, demineralization, color removal, and deacetylation of chitin (Uragami and Tokura, 2006). Nanoparticle is solid colloid that has a size in the range of 1-1000 nm (Tiyaboonchai, 2003).
FIGURE 1. Chemical Structure of Chitosan
Ionic gelation method is interesting because the process is simple, does not use organic solvent, and can be controlled easily. Gelation or gel formation is a merger or cross-linking of polymer chains to form a continuous threedimensional network and can trap water inside resulting compact and rigid structure (Fardiaz, 1989) as shown in Figure 2.
FIGURE 2 Electrostatic charge of Chitosan with TPP
During the process of storage, the chitosan nano-particle grows. In general, the growth of chitosan nanoparticles that occurred can be categorized into three types of processes (Wen, 2014). Following the results of the calculation and simplification to get the mechanism of the growth process, brief description of Wen models is described as follows : 1. Model 1 : Diffusion controlling ୈ Diffusion process occurs when the kinetics of particle surface took place very rapid ( > R). Particle ୩ growth is controlled by monomer adsorption into nanoparticle surface and the equation applied is : ࢊࡾ ࢊ࢚
ࢇ
ൌࡰ ቀ
ࢀ ࡾࢉ
െ ቁ ࡾ
(2)
Equation (2) is used to represent the adsorption controlling on the process of the growth of chitosan nanoparticle and is referred as model 2. 3. Model 3 : Adsorption and Diffusion Controlling It happens when adsorption and diffusion took control on the nanoparticle growth. If there is a case like this , the equation used is : ࢊࡾ ࢇ ൌ ࡰ ቀ െ ቁ ࢊ࢚
ቀ ାࡾቁࢀ ࡾࢉ
ࡾ
(3)
Equation (3) can be applied to represent the adsorption and diffusion controlling on the process of the growth of chitosan nanoparticle and is referred as model 3.
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RESEARCH METHODOLOGY Materials The main material was chitosan, medical grade powder, bought from PT. Biotech Surindo, Cirebon, West Java with deacetylation degree above 90%. Acetic acid as diluent of chitosan was glacial acetic acid p.a. from Merck. Crosslinker was Sodium Tripolyphosphate (TPP) p.a. from SIGMA-ALDRICH. Tween 80 used as surfactant was from MEDIKA. Apparatus The apparatus was simply such as magnetic strirrer, beaker glass 2L, and micropipet.
Method a.
Preparation of Chitosan Solution Every sample of chitosan powder, 2, 4, and 6 grams, was added into acetic acid 1% (v/v) gradually resulting concentration of 0.2%, 0.4%, and 0.6% chitosan respectively. The solution was stirred using magnetic stirrer till chitosan completely dissolved and homogenous. b. Preparation of Tween 80 Solution Tween 80 concentration used in the research was 0.1% (v/v). Tween 80 of 100 μL was diluted into 100 mL of distillated water. Tween 80 was then added into chitosan 0.2%, 0.4%, and 0.6% w/w respectively in amount of 250μL, 500 μL, and 750 μL. c. Preparation of TPP Solution The ratio between TPP and chitosan solution is 2:5 (v/v) based on Tsai et al, (2011) thus TPP solution should be prepared in the amount of 400 mL. TPP of 0.336 gram was dissolved into 400 mL distilled water then it was stirred using magnetic stirrer till formed homogenous solution. d. Preparation of Nano-chitosan solution Tween 80 solution as much as 250 PL was added into beaker glass of 2 L containing chitosan solution, using micropipette. The mixture was then stirred continuously for 30 minutes. Subsequently, TPP solution as much as 400 mL was added into the solution gradually. The mixture was stirred for another 1 hour until the cross-linking process takes place. After cross-linking, 100 mL of sample was analyzed to get the particle sizes. The experiments were conducted at 15qC and 30qC, with chitosan concentration of 0.2%, 0.4%, and 0.6%. The size of the particle was measured by Particle Size Analyzer (PSA).
RESULT AND DISCUSSION The Effect of Temperature and Chitosan Concentration on Particle Size The effect temperature and chitosan concentration on particle sizes during storage are presented in Figure 3.
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R(nm)
600 500 400 300 200 100 0
30 C 15 C
0
5
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20
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Time (Day)
R (nm)
(a) 800.00 700.00 600.00 500.00 400.00 300.00 200.00
30 C
0
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10
15
15 C
20
25
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Time (Day)
R (nm)
(b) 800.0 700.0 600.0 500.0 400.0 300.0 200.0 100.0 0.0
0.2%
0
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Time15(Day) (c)
0.4%
20
0.6%
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FIGURE 3. The effect of temperature on particle radius with the chitosan concentration (a) 0.2%, (b) 0.6% and (c) various chitosan concentration at 30oC
The interesting phenomena from Figure 3 is the similar range of nanoparticles size on temperature 15qC and 30qC within various concentration. It showed that temperature variance does not affect significantly the initial size of the nanoparticles produced as well as the growth of the nanoparticles during storage. On the other hand based on Figure 3 part c, initial chitosan concentration strongly affects initial size of the nanoparticles produced and the growth of the nanoparticles during storage.
The Mechanism of Nano-chitosan Particle Growth Process During storage, chitosan particles tend to grow and produce larger size. In this study particle growth occurred as indicated by changes in the size of the particle radius, but the growth is slow, so that the particle size is only slightly changed. To test the mechanism of nano size chitosan particle growth, the calculation results by model 1, 2 and 3 will compared with the experiment. The parameter value was calculated by comparing the results with experiment (visual ୈ inspection). Accordingly, it resulted the value of a = 150000 dan = 500. ୩
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300.0 R (nm)
200.0 R Data Model 1
100.0 0.0 0
5
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15 20 Waktu (Hari) (a)
25
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R (nm)
300.0 200.0 R Data
100.0 Note: Data on day 14 is not plotted caused by high deviation (outlier).
0.0 0
5
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15 20 Waktu (Hari) (b)
25
30
FIGURE 4 Comparison of the value of the radius (R) from experiment and simulation models (1, 2, and 3) at various time with 0.2% chitosan concentration at (a) 15qC and (b) 30qC
600.0
R (nm)
500.0 400.0 300.0 R Data Model 1 Model 2
200.0 100.0 0.0 0
5
10
15 20 Waktu (Hari)
25
30
FIGURE 5 Comparison of the value of the radius (R) from experiment and simulation models (1, 2, and 3) at various time with 0.4% chitosan concentration at room temperature
1000.0
R (nm)
800.0 600.0
R Data Model 1 Model 2 Model 3
400.0 200.0 0.0 0
5
10
15 Waktu (Hari) (a)
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20
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R (nm)
800.0 700.0 600.0 500.0 400.0 300.0 200.0 100.0 0.0
R Data Mod el 1 0
5
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20
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Time (Day) (b) FIGURE 6. Comparison of the value of the radius (R) from experiment and simulation models (1, 2, and 3) at various time with 0.6% chitosan concentration at (a) 15qC and (b) 30qC
Based on Figure 4 (a and b) and Figure 5 it was observed that simulation data are relatively close to the value of experiment data on every models (1, 2 and 3). However, if it is observed visually, model 2 and 3 are very close each other, ୈ while model 1 slightly deviates. Based on the value of obtained, model 2 is more appropriate to be used as the mechanism ୩
ୈ
of chitosan nanoparticle growth process. It is caused by the value of = 500 is greater than Rdata. Thus it can be concluded ୩ that adsorption controls the mechanism of chitosan nanoparticle growth. Figure 6 (a and b) give the results that are not much different with Figure 4, and Figure 5. Visually, simulation result for every models (1, 2, and 3) is close to the experiment. If it is examined closely, model 1 is more appropriate to be used. ୈ It is caused by the value of = 500 is smaller than Rdata. As a result, higher chitosan concentration which gave large ୩ initial particles sizes made the diffusion controls the mechanism of chitosan nanoparticle growth.
CONCLUSION AND ADVICE Based on the research and discussion above, it might be conclude and adviced that : The mechanism of chitosan nanoparticle growth process during storage on R < 500 nm is an adsorption controlling process, while on R > 500 nm is a diffusion controlling process. 2. Temperature variance (15qC and room temperature around 30qC) does not give significant effect on initial particle size and nanoparticle growth during storage. 3. The preparation of chitosan nanoparticle solution using ionic gelation method on 0.2%, 0.4%, and 0.6% chitosan concentration give the chitosan initial radius of 175.3 nm, 337.9 nm, and 643.3 nm respectively. ୈ 4. Curve fiting result gives the value of a = 150000 and = 500. ୩ 5. To obtain small particle size of chitosan nanoparticle, it recommended to use less chitosan concentration (0.2%) on the process of production using inonic gelation method. Nomenclature a = contants D = diffusivity coefficient (nm2/day) k = mass transfer coefficient (nm/day) Kb = Boltzman constants (J/molekul.K) R = particle radius (nm) Rc = nucleation radius (nm) T = temperature (K) t = time (day) 1.
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