79 Squalen Bull. of Mar. and Fish. Postharvest and Biotech. 13 (2) 2018, 79-84 www.bbp4b.litbang.kkp.go.id/squalen-bulletin Squalen Bulletin of Marine and Fisheries Postharvest and Biotechnology ISSN: 2089-5690 e-ISSN: 2406-9272 Copyright © 2018, Squalen BMFPB. Accreditation Number: 21/E/KPT/2018. DOI: http://dx.doi.org/10.15578/squalen.v13i2.297. PHYSICOCHEMICAL CHARACTERISTICS OF SODIUM ALGINATE EXTRACTED FROM Turbinaria sp. AND Sargassum sp. Rinta Kusumawati,1 Jamal Basmal1, and Bagus Bandol Utomo1 1)Research and Development Center for Marine and Fisheries Product Processing and Biotechnology Jl. KS. Tubun Petamburan VI, Jakarta 10260 Indonesia Abstract Characterization of sodium alginate extracted from two species of brown seaweeds (Turbinaria sp. and Sargassum sp.) harvested from Binuangeun Beach, Banten, has been conducted. The aim of the study was to evaluate physico- chemical characteristics (moisture, whiteness, viscosity, and functional groups) of sodium alginates extracted from Turbinaria sp. and Sargassum sp. Extraction was conducted in acid solution with the following steps: washing, acid extraction, bleaching, alginic acid conversion, sodium alginate conversion, dehydration, and drying. Each extraction was conducted in duplicate using approximately 1 kg of the dry raw materials (Turbinaria sp. and Sargassum sp.). Results of the analysis showed that the yield of sodium alginate powder extracted from Sargassum sp. was 24.56+0.56% (w/w) with moisture content of 12.69+1.24%; whiteness degree of 43.80+1.71%; and viscosity of 143.43+3.25 cPs, while the yield of sodium alginate powder extracted from Turbinaria sp. was 22.69+2.12% (w/w) with moisture content of 14.77+2.55%; whiteness degree of 23.77+0.68%; and viscosity of 133.67­+4.04 cPs. Meanwhile, the commercial sodium alginate was identified to have moisture content of 16.07+0.09%, whiteness degree of 29.37+0.55% and viscosity of 102.67+4.04 cPs. This indicates that physicochemical characteristics of sodium alginate extracted from Sargassum sp. is better than those extracted from Turbinaria sp. and commercial alginate since it had lower moisture content as well as higher whiteness degree and viscosity. Keywords: acid extraction, sodium alginate, Sargassum sp., Turbinaria sp. *Corresponding author. E-mail: tanjung.1979@gmail.com Article history: Received: 8 January 2018; Revised: 20 July 2018; Accepted: 25 August 2018 1. Introduction Alginate is a hydrocolloid extracted from brown seaweed. According to BPS (2018), Indonesia imported about 1,650 tons of alginate every year which was entirely supplied from imports. As much as 50% of the imported alginate was used in textile industry, 30% for food, 6% for paper production, 5% for welding rods production, and the other 5% for pharmaceutical purposes. Brown seaweed, as a raw material for alginate extraction, can be found in Indonesian waters. Two types of brown seaweeds which grow predominantly in Indonesia are Sargassum sp. and Turbinaria sp. However, not similar to Eucheuma cottonii which has been cultured successfully cultured in many areas in Indonesia, Turbinaria sp. and Sargassum sp. has not been cultured intensively (Purnomo et al, 2017). Therefore, in order to develop alginate industry in Indonesia, improving the method for alginate for alginate extraction to produce good quality product is very important, besides providing the high capital investment for the raw material sustainability. There had been several research reports on alginate extraction using acid followed by additional of formalin solution (Reyes-Tisnado, R.,Hernandez- Carmona, G., Montesinos, Y. E. R., Higuera, D. L. A., & Gutierrez, F.L. 2005, Jelynne, P., Tamayo, & Del Rosario, E.J., 2014, Viswanathan & Nallamuthu, 2014). Other methods were reported using acid solution for extraction and dehydrated using suitable solvent to form fibrous material (Murdinah et al, 2005; Husni, A., Subaryono, Pranoto, Y., Tazwir, & Ustadi. 2012, Latifi, A.M., Nejad, E.S., & Babavalian, H., 2015). Research for alginate production commonly used Sargassum sp. as a raw material, however, http://www.bbp4b.litbang.kkp.go.id/squalen-bulletin http://dx.doi.org/10.15578/squalen.v13i2.297. mailto:tanjung.1979@gmail.com 80 R. Kusumawati, J. Basmal, and B.S.B. Utomo/Squalen Bull. of Mar. and Fish. Postharvest and Biotech. 13 (2) 2018, 79-84 alginate can also actually be extracted from Turbinaria sp. Although the resource of Turbinaria sp. is still abundance in Indonesia, the experiments on alginate extraction in Indonesia from this species is still limited. Extraction using acid added with isopropyl alcohol (IPA) was chosen in in this study to avoid the use of formalin which is not recommended to be used in food processing. In addition, the use of IPA is able to speed up the drying process as well. Jothisaraswathi, S., Babu, B., and Rengasamy, R. (2006) reported that alginate extracted from ‘the leafy parts’ of the seaweed has the highest yield than that extracted from stem or entire thallus of Turbinaria conoides (J.Ag.) Kutz. However, it had the lowest viscosity, and molecular weight than alginate from S. binderi and S. baccularia (Chee, S., Wong, P., & Wong, C. , 2010). Viscosity and molecular weight are specific characteristics of alginate that can indicating the functional properties of alginate in commercial products. Beside the ratio of manuronate and guluronate, other properties such as moisture content, viscosity, whiteness, and functional groups are also important characteristics to determine the quality of alginate. The ai m of th e stu dy wa s to i dent i f y phy si co chem i cal chara cteri sti c s (m o i stu re, whiteness, viscosity, and functional groups) of sodium al gi nates ex tracted f rom Turbinaria sp. and Sargassum sp harvested from Binuangen Beach, Banten, Indonesia. 2. Materials and Method 2.1. Materials The study used Sargassum sp. and Turbinaria sp. collected from Binuangen beach, Banten, Indonesia, which were harvested during low tide condition. Other materials used were commercial sodium alginate (4mular Irving CA 92614), sodium alginate powder (Aldrich chemistry 9005-38-3), HCl, fresh water, Na2CO3, NaOCl, NaOH, C3H7OH, and chemicals for Fourier-transform Infrared (FTIR) spectroscopy analysis. 2.2. Method 2.2.1.Samples preparation The brown seaweeds (Sargassum sp. and Turbinaria sp.) were washed with fresh water followed by cleaning the samples. The samples were sundried and packed hermetically in plastic bag, then transported to laboratory and stored at room temperature for further experiment. 2.2.2.Sodium alginate extraction Sodium alginate extraction was conducted based on Murdinah et al (2005). Kg of dry seaweed, with having moisture content 11.96+0.03%, were soaked in 1% (v/v) HCl with ratio of dry seaweed to HCl solution was 1:30 (w/v), and suspended for 1 hour then neutralized by flowing fresh water. Sodium alginate extraction using neutral raw material was conducted in 2% (w/v) Na2CO3 at 60–70°C for 2 hour, with proportion of dry seaweed to Na2CO3 solution was 1:30 (w/v).The suspended material from previous extraction step was re-extracted, filtered and the supernatant was mixed with the first extract then bleached by adding 4% (v/v) NaOCl slowly for 30 minutes. The alginate solution obtained from extraction process was then acidified by adding 10% (v/v) HCl until the acidity of the solution reached a pH of 2 – 3 and then filtered by using vibrator screen. The solution was converted into sodium alginate by adding 10% (w/v) NaOH solution until the pH of the solution was 7 – 8. The sodium alginate was then dehydrated using C3H7OH with the proportion of sodium alginate to C3H7OH was 1:2 (v/v) followed by smooth stirring for 30 minutes until solid state of sodium alginate was formed. The solid state of the sodium alginate was dried at 50°C for 10 hours and then milled into powder using 100 mesh of sieve plate. This extraction procedure was applied for both Sargassum sp. and Turbinaria sp. 2.2.3. Mosture content analysis Mosture content was analyzed following SNI 2534- 1:2015 analysis procedure (BSN, 2015). As much as +2 g sodium alginate powder, were dried for 18-24 hours in the oven at 1050C, then cooled in a desiccator for 30 minutes and weighed after temperature of the samples reach 250C. The value of moisture content is the percentage of the weight ratio of the water evaporated to the initial sample weight. 2.2.4. Viscosity analysis Viscosity was measured based on Brookfiled viscometer manual book using Brookfiled LVT model viscometer. 7.5 grams of sodium alginate powder were dissolved in 500 ml of distilled water then stirred slowly at 800C for 30 minutes. Analysis was conducted using a spindle number 2 with 60 rpm speed rotation and keeping the temperature of the solution stable. The stable number indicated by the tool was then multiplied by the conversion factor to obtain a viscosity value. 81 2.2.5. Whiteness degree analysis W hiteness degree was measured based on whiteness meter manual procedure using Kett whiteness meter. Measurement using white paper as a standard and powder samples. After measuring absorption of the standard, absorption of the sample measured by filling the sample into whiteness meter chamber until full and compact. The whiteness value indicated by the tool is the percentage of the trend toward the white color of the sample. 2.2.6. Functional groups analysis The FTIR (Perkin-Elmer 577) was used to analyze of functional groups of the sample using procedure manual for KBr pellet method and M-80 spedcord. One milligram of sample was crushed with 200 mg KBr until it was homogeneous. The powder was pressed into a thin tablet and placed in a sample pan. Infrared uptake was measured at a waveleth of 400- 4,000 cm-1. 2.2.7. Data analysis The experimental design used in this study was Com pl ete l y R andom i zed Desi g n wi th th ree replications. Data obtained were analyzed using MS- excel program. 3. Result and Discussion The yield of sodium alginate extracted from Sargassum sp. was 24.56%, higher than Turbinaria sp. which was only 14.77%. It was probably due to the difference in the thallus hardness. The thallus of Sargassum sp. was softer than that of Turbinaria sp. Stiger, Deslandes, and Payr (2004) found that morphological point of view affected the yield of sodium alginate extract. Due to their thallus texture difference, extracting alginate from Sargassum sp is easier than from Turbinaria sp, thus the yield of alginate which can be extracted from Sargassum sp was also higher than that from Turbinaria sp, Stiger et al (2004) also found that morphologically, Sargassum sp has softer and more flexible thallus than Turbinaria sp. which was strongly embeded to corral reef. Extracting alginate from Turbinaria probably needs additional treatment to produce an optimal or even maximum yield. It could be done by using higher concentration of extracting solution or temperature. However, further study related to this needs to be conducted. For commercial production, Sargassum sp. as raw material has a better opportunity to be developed because this seaweed is more easily found in Indonesian coastal area. However, this type of seaweed has not been cultivated commercialy, even though the cul ti v ati on ex peri ment conducted i n 2000 at Bi n uang eun B each was qui t e suc cessf ul (Kusumawati, 2011). According to Soegiarto, A., Soelistijo, Atmadja, W.S. &Mubarak, H. (1978), yield of alginate is depent on the type of seaweed, the environmental condition and season, whereas Budiyanto and Djazuliu (1997) mentioned that the yield o alginate depended on light intensity, water current, nutritional condition, the age of the seaweed, method of handling as well as extraction techniques. Yulianto (1997) stated that pre-treatment which was conducted 24.56+0.56 12.69+1.24 43.80+1.71 143.43+3.25 22.69+2.12 14.77+ 2.55 23.77+0.68 133.67+4.04 16.07+0.09 29.37+0.55 102.67+4.04 0.00 20.00 40.00 60.00 80.00 10 0.00 12 0.00 14 0.00 16 0.00 Yield (%) W ater Content (%) W hiteness (%) Viscosity (cPs) Sargassum sp. Tu rbinaria sp. Alginate Comme rcial Figure 1. Yield and characteristics of alginate extracted from Sargassum sp. and Turbinaria sp. R. Kusumawati, J. Basmal, and B.S.B. Utomo/Squalen Bull. of Mar. and Fish. Postharvest and Biotech. 13 (2) 2018, 79-84 82 before extraction was v ery important factor in determining the quality of the extracted alginate. Water content affected the shelf life of the product. Nasir et.al (2013) reported that the shelf life of powder are depending very much on the water content which affects especially on microbiological activity and insect infestation. The higher the moisture content the faster is the growth of fungi and also insects infestation, means that the lower the moisture content of alginate powder the longer is its shelf life. Sodium alginate powder extracted from Sargassum sp. had moisture content of 12.69+1.24%, lower than that from Turbinaria sp., which was 14.77+2.55%. Since the structure of sodium alginate extracted from both brown seaweed might has some differences, the position of water trapped inside the polymer of alginate may also be different. It needs more research regarding the type and characteristics of the polymer in relation to water trapped in the products. Comparing to commercial sodium alginate powder which had moisture content of 16.07+0.09%, the al ginate extracted from Sargassum sp. and Turbinaria sp. have lower moisture content, meaning that those products might have longer shelf life due to lower microbial activity and growth of fungi. Whiteness degree of alginate powder is correlated with the use of alginate in food and pharmaceutical industry. The high degree of alginate powder ’s brightness or whiteness, would give browning effect to the natural color of the main material in formulation. In this study, the whiteness degree of sodium alginate from Sargassum sp. was higher than alginate from Turbinaria sp. (43.80+1.71 and 23.77+0.68%)as well as sodium alginate commercial (29.37+0.55%). This phenomenon showed similar contend to the moisture content. Similarly, viscosity of alginate powder is associated with its function as emulsifier or binder in product formulation in food and pharmaceutical industry. Until recently, Indonesia is still importing alginate for this purpose because it requires a specific viscosity of alginate. Production of alginate powder that meeting the specification needed can support the industrial demand and reduce alginate imports. The viscosity of sodium alginate from Sargassum sp (143.43+3.25 cPs) was higher than alginate from Turbinaria sp. (133.67+4.04 cPs). Both of sodium alginates have hi gher v iscosi ty v alues than sodium al ginate commercial (102.67+4.04 cPs). Viscosity is important phy si cal cha racte ri sti c f o r al g i nate , si nce concentrations of alginate in water causing thickening effect during stirring process (McHugh, 1987). In order to get a thickening effect in a product a small amount of high viscosity alginate can simply be added into formulation. Quantitative information to describe the functional groups and composition of sodium alginate is very important. This prarameter was analyzed using FT-IR spectroscopy (Figure 2). A band at around 3,400 cm- 1 was attributed to O-H stretching vibration, a band at around 2,900 and 1,600 cm-1 was attributed to C-H stretching to carboxylate group, while a band at around 1,400 and 1,600 cm-1 indicated the presence of carbonyl group (Daemi & Barikani (2012), Gholamipoor et al. (2013), and Liu et al. (2016)). the absorbance values of sodium alginate extracted from Sargassum sp. and Turbinaria sp. were not at the same band, indicating that they had different ability in absorbing Figure 2. FT-IR spectrum of sodium alginic standard, and sodium alginic extracted from Sargassum sp. and Turbinaria sp. R. Kusumawati, J. Basmal, and B.S.B. Utomo/Squalen Bull. of Mar. and Fish. Postharvest and Biotech. 13 (2) 2018, 79-84 83 infrared light. Table 1 is the summary of the absorbance band characteristics around 3400 – 1000 cm1. Based on FTIR analysis, alginate extracted from Sargassum sp. and Turbinaria sp. had a characteristic of high absorbance intensity at specific frequency. These specific frequency was very similar to those of the alginate standard used in the experiment, ie. 3,446.38; 2,928.41; 1,628.41; 1.628; 1,420.78, and 1.033.18 cm-1. Some frequency were not exactly the same with the standard, they were slightly different (a little bid higher or lower) due to atomic bounding interaction. Pereira, L, Gheda, S.F., & Ribeiro-Claro, J.A. (2013) reported that manuronate and guluronate ratio (M/G) of alginate can be tentatively estimated at band value around 1,030 to 1,080 cm-1 in infrared spectra. The result showed that intense broad band of sodium alginate extracted from Turbinaria sp. centered at around 1,030 cm-1 of band absorbance, might indicate that the sample contain a lot of guluronate acid (Pereira et al, 2013). The band shifting seems to happen on sodium alginate extracted from Sargassum sp. which may be caused due to the existence of impurities. To determine which type and quantity of impurities affected the band shifting, further study need to be conducted. 4. Conclusion Sodium alginate extracted from Sargassum sp. has yield of 24.56+0.56%, moisture content of Sodium Alginate Sodium Alginate extracted Sodium Alginate extracted Assignments Standard From Sargassum sp. From Sargassum sp. (Fertah et al, 2017) (cm-1) (cm-1) (cm-1) 1 3,446.38 3,446.08 3,446.70 Hydrogen bonded O-H stretching vibration 2 2,928.41 2,921.50 2,928.81 C-H stretching vibration 3 1,628.00 1,636.21 1,630.35 Asymmetric stretching of carboxylate O-C-O vibration 4 1,420.78 1,417.22 1,404.81 Symmetric stretching vibration of the carboxylate group 5 1,033.18 1,034.44 1,115.37 Due to C-O stretching vibrations No Table 1. Absorbance band of sodium alginate standard, sodium alginate extracted from Sargassum sp. and Turbinaria sp. 12.69+1.24%, whiteness degree of 43.80+1.71% and viscosity of 143.43+3.25 cPs; while sodium alginate ex t racte d f ro m Tur binari a sp. has yi el d of 22.69+2.12%, water content of 14.77+2.55%, whiteness degree of 23.77+0.68% and viscosity of 133.67+4.04 cPs. Meanwhile, the commercial sodium alginate was identified to have moisture content of 16.07+0.09%, whiteness meter of 29.37+0.55% and viscosity of 102.67+4.04 cPs The physicochemical characteristics of sodium alginate extracted from Sargassum sp. is better than those extracted from Turbinaria sp. and commercial alginate since it had lower moisture content as well as higher whiteness degree and viscosity. References Badan Pusat Statistik (2018). Expor dan Impor: Tabel Impor Menurut Komiditi, Tahun 2017. Badan Standardisasi Nasional. (2015). 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