Functional Characteristics of Cookies Containing Snakehead (Ophiocephalus striatus) Fish Protein Concentrate Fortified with Chlorella sp.

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International Journal of Oceans and Oceanography ISSN 0973-2667 Volume 12, Number 1 (2018), pp. 43-52 Research India Publications http://www.ripublication.com Functional Characteristics of Cookies Containing Snakehead (Ophiocephalus striatus) Fish Protein Concentrate Fortified with Chlorella sp. Dewita*, Syahrul, and Desmelati Department of Aquatic Product Technology, Faculty of Fisheries and Marine Science, University of Riau *Corresponding author Abstract Microalgae, or better known as phytoplankton, have been introduced as a new food source, because it has excellent nutritional content which is even better than other foods which are commonly consumed by Indonesia people. Among any types of microalgae, Chlorella sp. is one of potential microalgae which is well known as a source of food nutrition. This study aimed to produce raw materials for food industries in the form of concentrates (powder) of Chorella sp. biomass, to analyze its characteristics, and to assess characteristics of cookies containing snakehead fish protein concentrate (Ophiocephalus striatus Bloch, 1793) fortified with Chlorella sp. Microalgae powder was applied in food product in the form of cookies, thus, suitable for children under five years old. This methods research in a several phase,, make a snakehead protein concentrate (Ophiocephalus striatus) fortified Chlorella sp, chemical compostition by AOAC Amino acids method by HPLC, vitamine and minerals by HPLC and AAS, and microbiology analysis by total plate count. Acceptance panelist about product by sensory test. The results showed that Chlorella sp. contained 44.67 % protein, 19.84 % fat, 21.00 % carbohydrates, 4.92 % fiber, and very low content of heavy metals (0.000 277 64 mg L 1 Pb and 0.000 234 12 mg L 1 Cd). Organoleptic test on the cookies shows the acceptance by panelists (82.50 % to 98.75 %) in which the cookies contained 21.65 % protein, 12.72 % fat, 54.83 % carbohydrates, 3.68 % fiber, and no heavy metals detected. Keywords: Chlorella sp. powder, functional cookies, snakehead fish protein concentrate

44 Dewita, Syahrul, and Desmelati INTRODUCTION Indonesian people have been familiar with utilizing natural food sources, especially those which are commonly known. On the other hand, they tend to be less interested in using food sources which people are not familiar with, or even a completely new food sources. Thus, the act of socialization and counseling to introduce new alternative food sources is needed. Microalgae, or better known as phytoplankton, has been introduced as a new food source, but people are not very enthusiastic to use this food source although microalgae nutritional value is very good, even better than other foods. Among the types of microalgae, Spirulina sp., Chlorella sp., and Dunaliella sp. are well-known as potential food source. Some research has been conducted on snakehead and microalgae, such as biomass utilization of Spirulina platensis powder in the manufacture of lozenges (Saputra et al., 2014), extraction of bioactive compounds as natural antioxidants from fresh Spirulina platensis using different solvents (Firdayani et al., 2015), growth and extracelluler polysaccaride production of Porphyridium cruentum (S.F. Gray) Nägeli, 1849 in various photoperiod (Prasetyo et al., 2015), phycocyanin extraction from Spirulina platensis and its antimalarial activity in-vitro (Wulandari et al., 2016). Previously, research report about Siam patin protein concentrate on amplang snack and instant sago noddle as leading product in Riau, and innovative cookies product from patin protein concentrate have been published (Dewita & Syahrul, 2014; Dewita & Syahrul, 2015). Suwandi et al., (2014) also published a report about proportion and chemical composition of snakehead. Information about microalgae processing technology to produce ready-to-eat food are still limited, thus, most resulting products do not meet consumer requirements as food source. Information about microalgae processing technology to produce ready-to-eat food are still limited, therefore, most resulting products do not meet consumer requirements as food source. Therefore, it is necessary to investigate the characteristics of cookies containing snakehead fish protein concentrate fortified with Chlorella sp. Microalgae Chlorella sp. has good potential as source of natural ingredients that can be used for groceries and also to increase the nutrients in food for humans. Utilization of microalgae processing technology to produce ready-to-eat food is still limited, therefore, most resulting products do not meet consumer requirements as food source, therefore, for further development, technology improvement is required to make Chlorella sp. as food fortification ingredient. Therefore, fortification of Chlorella sp. into cookies containing snakehead fish protein concentrate which was acceptable by panelists and analysis on the characteristics of the cookies were performed in this study. The purpose of this study was to produce food industry raw materials in the form of concentrates (powder) of the biomass of Chlorella sp. and snakehead fish protein concentrate and to investigate the characteristics of cookies containing snakehead fish protein concentrate fortified with Chlorella sp. Theoretically, this

Functional Characteristics of Cookies Containing Snakehead 45 research can be used as information and contribution to the development of cookies as a nutritious food product. MATERIALS AND METHODS Materials and Equipments The raw material used in the study was snakehead fish (Ophiocephalus striatus Bloch, 1793), wheat flour, refined sugar, butter, egg yolks and Chlorella sp. powder. Food grade chemicals, such as isopropyl alcohol and NaHCO3, were used for cookies production. Research Methods Cookies containing snakehead fish protein concentrate fortified with Chlorella sp. powder was made based on formula in Table 1. Quality parameters were tested by performing chemical analysis (proximate), microbiological analysis (total plate count, TPC) and organoleptic test (Test A) with 80 respondents as untrained panelists. This research was conducted through the following steps, i.e. processing snakehead fish protein concentrate (Ophiocephalus striatus), and processing cookies containing snakehead fish protein concentrate fortified with Chlorella sp. powder. TABLE 1 The formulations in the manufacture of cork-fortified cookies fish protein concentrate Chlorella sp. Composition Weight (g) Snakehead fish protein concentrate 10 Wheat flour 100 Sugar powder 120 Margarine 100 Yellow egg 4 Chlorella sp. powder 2 Production of Snakehead Fish Protein Concentrate Snakehead fish protein concentrate was produced based on previously developed method by Dewita et al. (2011). Fresh snakehead fish (1 kg to 1.5 kg) was collected and transported to laboratory of Fishery Product Technology, Faculty of Fisheries, University of Riau. The fish was cut into fillet, the fish scales were removed, then they were cut into small pieces, grinded with a meat grinding machine (food processor), and added with 0.5 % (w/w) salt. The fish was crushed, steamed for 30

46 Dewita, Syahrul, and Desmelati min, and pressed to reduce water and fat content. Next, in order to form a paste, 0.5 N of NaOHCO3 was added until isoelectric ph was achieved Solvent extraction was performed using isopropyl alcohol 1:3 (v/v) and followed by extraction for 10 h to form a precipitate or residue. Subsequently, the sample was dried at 40 C for 15 h in cabinet dyer, and then blended and sieved to obtain snakehead fish protein concentrate powder. Afterwards, the powder was packed in aluminum foil and stored in a cold storage. Snakehead Fish Protein Concentrate The steps of making cookies were performed as follows. Sugar powder and egg yolks were mixed slowly using mixer to form a smooth dough. Then, Chlorella sp. powder and snakehead fish protein concentrate were gradually added into the mixture to form a fine dough. Furthermore, the dough was molded into a uniform size using a mold. Molded dough was prepared on an aluminum pan and put in the oven with a temperature of 160 C for 10 min to 15 min until cooked. Eventually, cookies were cooled in room temperature to harden the texture due to the existence of sugar and fat. Data Analysis Data were tabulated, processed and analyzed descriptively. Data result were described and compared with literature. RESULTS AND DISCUSSION Chlorella Powder Chemical analysis result on Chlorella sp. powder can be seen in Table 2. Table 2 shows that the powder of Chlorella sp. contained a high nutrient content, especially protein, fat and carbohydrates. Moreover, an extremely low amount of heavy metal content was detected. Thus, Chlorella sp. powder is very safe for use as food ingredients. According to Kawaroe et al. (2012) besides having a high nutritional value, Chlorella sp. also contains chlorellin, which is an antibiotic compound that can inhibit the growth of pathogenic bacteria.

Functional Characteristics of Cookies Containing Snakehead 47 TABLE 2 The proximate composition of Chlorella sp. powder Composition Percentage (%) Protein Fat Carbohydrate Fiber Mineral and vitamin Heavy metal : Pb (mg L 1 ) Cd (mg L 1 ) 44.67 19. 84 21.00 4.92 957. 0.000 234 0.000 277 TABLE 3 The proximate composition of fortified cookies Composition Fortified Cookies PAG Standard (1972) Water (%) 7.12 5 to 10 Fat (%) 12.72 Min 10 Protein (%) 21. 65 Min 15 Abu (%) 3.65 Max 5 Fiber (%) 3.68 Max 5 Carbohydrates (%) 54.83 - Calories (cal) 4.960 1.100 to 1.300 Fortification of Chlorella sp. Powder to Cookies Containing Snakehead Fish Protein Concentrate Based on organoleptic test result, 97 % respondents which consisted of toddler at 4 yr to 5 yr of age were able to accept the cookies. This indicates that the cookies may be accepted or liked by toddlers. Proximate analysis result of fortified cookies is presented in Table 3. Table 3 shows that the value of the proximate composition fortified cookies meet the standards of Protein Advisory Group (PAG, 1972) for toddler food. Furthermore, the results of vitamin and mineral analysis of fortified cookies can be seen in Table 4. Vitamin A content

48 Dewita, Syahrul, and Desmelati TABLE 4. Vitamin and mineral content of fortified cookies Nutrition Fortified cookies Nutritional adequacy rate* Vit. A (UI) 223 160 Vit. C (mg) 16.70 17 Calcium (mg) 317 190 Fe (mg) 3.48 7 Zinc (mg) 2.67 2.7 *Ministry of health TABLE 5. The rate of consumer acceptance of cookies fish protein concentrate cork fortified Chlorella sp. Criteria Parameter Look Smell Taste Texture Amount % Amount % Amount % Amount % Very Like 30 37.5 36 45 27 33.75 35 43,75 Like 47 58.75 43 53.75 39 48.75 42 52.52 Dislike 3 3.75 1 1.25 4 5 3 3.75 Hate 0 0 0 0 0 0 0 0 of the fortified cookies was higher than in standard cookies. On the contrary, vitamin C and zinc content in the fortified cookies meet the standard of nutritional adequacy rate. According to Erick (2008), vitamin C is useful to increase the absorption of iron and repair tissue integrity. On the other hand, calcium content of the fortified cookies was higher than nutritional adequacy rate. However, Fe content shows different result in which Fe content in the fortified cookies was lower than nutritional adequacy rate (< 7 mg). Ca content assists in the formation of bones and teeth of the fetus, while iron is an essential mineral for the process of neurogenesis, myelination nerve and brain cell differentiation of the fetus during pregnancy. Zn content in the fortified cookies meet the nutritional adequacy rate. Zn acts as an enzyme cofactor for protein production amino acid essential) and transferrin which brings Fe and Zn into the blood and then distributes to the bone marrow as well as the production blood cell (Erick, 2008).

Functional Characteristics of Cookies Containing Snakehead 49 Sensory Test Sensory test was performed using 80 panelists. The result is presented in Table 5. In Table 5 can be seen cookies value for really like 96.20 %, odor by 98.75 %. Taste (82.50 %) and texture (96.25 %). Thus the test as a favorite of the panelists can be like (82.50 % to 98.75 %), caused sugar formula gave a sweet taste of eggs and butter add texture. In addition become to crunchy and soft (Sudha et al., 2007). Microbiology Analysis Microbiological parameter is one of the tools to determine the condition of sanitation and hygiene in cookies production. Based on the result in Table 6, the fortified cookies meet the requirement from Food and Drug Supervisory Agency (BPOM). TABLE 6. Microbological test cookies fish protein concentrate snakehead Parameter Fortified cookies BPOM Total bacteria Aerobic (ALT) 4.5 10 3 1.0 10 5 Escherichia coli < 2 < 3 Staphylococcus aureus < 2 < 2 Salmonella Negative Negative Therefore, as snack food, the fortified cookies is safe for children under 5 yr old. Moreover, during socialization of cookies production, audience was able to consume all cookies. Amino Acid Profile The content of essential amino acids of fortified cookies is presented in Table 7. Table 7 shows that the content of essential amino acids in fortified cookies meet AAE standard on essential amino acids intake per day. All essential amino acids contained in the cookies have their respective functions. As stated by Erick (2008), methionine is useful to help eliminate toxic substances in the liver, stimulate liver regeneration and reduce blood cholesterol levels; whereas leucine helps the breakdown of muscle protein and recovery of broken bones.

50 Dewita, Syahrul, and Desmelati TABLE 7. Essential amino acids content Essential amino acids Fortified cookies (%) AAE (%) Threonine 2.21 18. Methionine 0.96 1.44 Valine 3.26 2.7 Phenilalanine 2.23 1.8 Isoleusine 3.28 2.7 Leusine 4.42 3.06 Lysine 3.22 2.7 Furthermore, it is stated that isoleucine is needed in the production and storage of protein by the body and the formation of hemoglobin. It also improves the function of thymus and pituitary gland for optimal growth, maintains nitrogen balance in the body, and stabilizes blood sugar levels. The formation of other non-essential amino acids and hemoglobin is also assisted by isoleucine. Phenylalanine is needed to improve mood, endocrine levels and aphrodisiac (Beard, 2008). Other essential aminco acids like valine is also useful for growth, nervous system and digestion. Valine is also good to manage neuromuscular disorder, mental and emotional disorder, insomnia and anxiety. It also stimulates muscle coordination, repairs damaged tissues, and maintains nitrogen balance in the body (Saputra, 2014). CONCLUSION The fortified cookies was round in shape, and the sensory parameter such as aroma, taste, appearance, color resembled cookies in general. Organoleptic test results shows acceptance of the cookies by proximate composition of the cookies met the requirement for toddler food from PAG standard, as well as content of vitamin A and C, calcium and zinc which met the requirement of cookies Recommended Dietary Allowance (RDA). The result form microbiological test and essential amino acids profile also met the requirement from BPOM and AAE, respectively. ACKNOWLEDGEMENT Support from Taufik Hidayat.is greatly acknowledged.

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