The study aimed to evaluate the addition of banana blossom (12.5, 25, 37.5, and 50% w/w) on nutritional quality, histamine content, bacterial count, and sensory characteristic in the fish floss prepared from little tuna (Euthynnus affinis). The crude protein content, essential amino acids, lipid, and polyunsaturated fatty acids (PUFA) steadily decreased (p <0.05), while the crude fibre, carbohydrate, and ash components of the tuna floss, increased significantly (p <0.05) with increasing levels of banana blossom. The contents of protein, fat, ash, fibre, carbohydrate, and moisture ranged 28.13 – 30.27%, 14.79 – 18.02%, 4.45 – 5.68%, 2.6 – 3.5%, 27.81 – 31.01, and 16.45 – 17.39%, respectively, and most of them met the Indonesian National Standard. For essential and non-essential amino acids, the level varied about 102.82 mg.g-1 to 206.76 mg.g-1 and 79.71 mg.g-1 to 138.76 mg.g-1, respectively in the treated tuna flosses. Moreover, ranging 13.72 – 16.29% of PUFA was found in all treated flosses. The most significant effect was found in the histamine levels of the tuna flosses, especially in the 50% added floss sample. Moreover, bacterial counts and heavy metals content were lower than the maximum limits regulated by the Indonesian National Standard. For sensory evaluation, the banana blossom-added samples significantly increased (p >0.05) the acceptability score for all attributes assessed. Taken together, the tuna floss added with 37.5% of banana blossom may be potentially developed as a low-histamine tuna-based product with high ffibre andEPA+DHA, as well as highly acceptable for consumers.
Keywords: floss; little tuna; banana blossom; incorporation; characterizations
INTRODUCTION
Banana blossom (Musa acuminata), also commonly called banana flower, is considered as a leftover product after cultivation, which is widely consumed as a vegetable in the Southeast Asian countries, including Indonesia, Malaysia, Thailand, and the Philippines (Liyanage et al.,2016;Wahab, Ismail and Abidin, 2020). Banana blossom contains high nutritional quality, especially dietary fibre. According to Sharma, Shukla and Golani (2019), the banana blossom (100g) contained dietary fibre (5.74g), protein (1.6g), fat (0.6g), carbohydrate (5.7g), calcium (73.3mg), phosphorous (56.4mg) and vitamin E (1.07mg). Dietary fibre plays an essential role in lowering serum cholesterol levels, preventing obesity, normalizing blood glucose and insulin levels (Bhaskar et al., 2012;Elaveniya and Jayamuthunagai, 2014). In addition, banana blossom possesses some bioactive components, such as vitamin C, saponin, tannin, flavonoid, alphatocopherol, and myoinositol phosphates, which could promote health benefits (Somsub et al., 2008;Sheng et al., 2010). Thus, due to being rich in a range of nutrients and biologically active components, the banana blossom has gained considerable attraction as an alternative source of functional food ingredients.
Abon, also known as shredded fish or fish floss, is one of the traditional dried meat products popular among Indonesia and the Asian community. It is recognized by different local names such as serunding in Malaysia, mahu in the Philippines, moo yong in Thailand, thitheotieu in Vietnam, and rousing in China (Hang, 2015). Popular raw materials for producing floss are beef and chicken; however, some fish species are also suitable materials for shredded meat processing. The processing of floss product generally begins with steaming of washed meat until it is tender. The steamed meat is then shredded finely and mixed with some formulated spices, followed by adding coconut milk. After that, the mixture is fried and stirred continuously under heat until the mixture is dry. Finally, excessive oil in the dried meat product is then separated and removed automatically (Huda et al., 2012). Shredded meat is consumed as part of a daily dish or served as filler of lemper (traditional food made of glutinous rice and wrapped in a banana leaf) (Suryani, Abdurrachim and Alindah, 2016). As a dried meat product, it contains a range of nutrients, including protein/amino acids, fat/fatty acids, and ash/mineral. Protein and fat found in meat floss products are generally high at around 23.99 – 32.93% and 18.31 – 32.30%, respectively (Huda et al., 2012;Wazir et al., 2019;Fahmi and Purnamayati, 2020). A high level of fat is mainly caused by cooking oil absorbed during frying. As reported by Wazir et al. (2019), the meat floss contained approximately 87.73 – 91.65% of saturated fatty acids. For ash value, it contains around 3.17 – 6.67%. Among raw materials, a meat floss made of fish is much preferred by many people following the study from Huda et al. (2012), which revealed that fish floss has the highest score in overall sensory parameters compared to other meat floss samples.
Little tuna (Euthynnusaffinis) belongs to the Scombridae family that is categorized as a medium-sized tuna (Ahmed et al., 2015). It is one of the most commercially important marine fish species in Indonesia. According to the Ministry of Marine and Fisheries Affairs (2021), the total production of little tuna significantly increased around 61.27%, from 366,900 tonnes in 2010 to 592,056 tonnes in 2019. In terms of nutritional value, little tuna is rich in protein and contains the amount of polyunsaturated fatty acids (PUFA). Kannaiyan et al. (2019) reported the protein content found in the white and dark muscle of little tuna was around 23.15% and 23.12%, respectively, while PUFA was around 51.86% and 55.87% in the white and dark muscle, respectively. Due to nutrients rich, little tuna is extensively employed as an essential raw material for sashimi and canned products in the seafood processing industry. Also, other diversifications from little tuna have been developed, including shredded meat, nugget, dumplings, fish ball, and fish cakes (Suprayitno, Adi and Sulistiyati, 2016;Hizbullah et al., 2020). Fish floss derived from little tuna or other fish species mainly uses muscle part with adding some condiments. As a result, it has high protein content, but low dietary fibre. Moreover, histamine content would be high in the end-product because of the raw materials prepared particularly from the dark muscle of tuna and it could give rise to food-borne poisoning (Lee et al., 2016;Colombo et al., 2018). Therefore, there has been increasing interest in formulating food products with functional ingredients to deal with the unbalanced nutrition and undesirable component. Previous studies reported different quality aspects of banana blossom added nugget, noodle, biscuit, and meat floss (Wahab, Ismail and Abidin, 2020;Elaveniya and Jayamuthunagai, 2014;Komal and Kaur, 2019;Novidiyanto et al., 2020;Puspita, Kartikaningsih and Dayuti, 2019). Among them, the added banana blossom could elevate the amount of dietary fibre, antioxidant, and other functional properties.
Scientific hypothesis
There is a lack of information regarding the use of banana blossom incorporated with tuna floss in increasing fibre content and decreasing histamine levels. Therefore, this study hypothesized that incorporating little tuna floss with banana blossom may give favorable nutritional features, particularly enhancing fibre content and lowering histamine levels. Banana blossom was added up to 500 g.kg-1 in the formulation of little tuna floss. Nutritional aspects such as chemical composition, amino acids, fatty acids, and dietary fibre contents of the formulated little tuna floss were analyzed. Heavy metals content as safely required in the food developments was evaluated. The histamine content and pathogenic bacteria test were also determined. In addition, the sensory properties of the little tuna floss were studied to provide a basis for consumer acceptance and commercial applications. This study may stimulate further study in developing healthy fish floss products for nutritional and functional applications.
MATERIAL AND METHODOLOGYSamples
Little tuna (E. affinis) used in this study was obtained from Sedang Biru fish market (Malang, East Java, Indonesia). The size of little tuna samples was approximately 202.8 ±3.3 g and 27.6 ±1.9 cm for weight and length, respectively. Banana blossom (M. acuminata) was purchased from a local market (Malang, East Java, Indonesia), and its weight was around 2.2 ±0.2 kg.
Chemicals
All chemicals and reagents used were of analytical grade. H2SO4, petroleum ether and nitric acid (Merck, Germany) were supplied from a local supplier. Kjeldahl catalyst selenium tablet (Fisher Chemical, USA).
Bacteria and biological material
Bacteria used, including Escherichia coli, Salmonella sp. and Staphylococcus aureus were obtained from the Indonesian Culture Collection (InaCC), Research Center for Biology, Indonesian Institute of Sciences (LIPI). Microbial media used in a recent study were Oxoid (Basingstoke, UK)-based Nutrient agar (NA), MacConkey agar (MCA), Rappaport-Vassiliadis broth (RV), Xylose Lysine Deoxycholate (XLD) agar and Baird Parker agar (BPA).
Instruments
The instrument used consists of the oven (M720, Germany), Soxtec 2050 (FOSS Analytical, Denmark), ultra-pressure liquid chromatography (UPLC) (Waters, US), gas chromatography (GC) (Agilent Technologies, California, US), High-Performance Liquid Chromatography (HPLC) (Agilent Technologies, California, US) and Atomic Absorption Spectrophotometer (AAS) (GBC Scientific Equipment, USA).
Description of the experimentSample preparation
Around 15 kg of little tuna samples were purchased from Sendang Biru fish market (Malang, East Java, Indonesia). Fish samples were kept under cold conditions in an insulated box with ice during transportation (around 2-3 h). Upon arrival, the little tuna samples were washed in running water and were then filleted manually using a sharp knife. The filleted samples, including white and dark muscles, were stored in a freezer until use. For banana blossom, about 5 pieces of samples used were was obtained from a local market (Malang, East Java, Indonesia). Due to fresh banana blossom samples used, the detailed preparation was described in the following procedure for addition into little tuna floss.
Preparation of little tuna floss incorporated with banana blossom
Table 1 presents the distinct formulations for all little tuna floss samples. The banana blossoms added with banana blossom in preparing little tuna floss products were 12.5% to 50% based on the weight of the whole little tuna sample (Figure 1). The little tuna meat used consists of white and dark muscles with distinct proportions i.e., 80% and 20%, respectively. These treatments were chosen in compliance with the formulated proportion for fish floss recommended by the previous study from Puspita, Kartikaningsih and Dayuti (2019). The control little tuna floss (0%) was used as a control and compared to the treated little tuna floss samples (12.5 – 50%) to evaluate the distinct characteristics of the formulation process and its corresponding attributes on the treated little tuna floss produced. During preparation, the thawed little tuna muscles were steamed at 100°C for 15 min (Halco, Indonesia). Separately, the banana flowers were cut into small sizes and then steamed at the same condition of fish treatment. After the steaming process, the steamed tuna was shredded using a fork and all ingredients used were thoroughly mixed into the pan. Afterward, the mixture samples were fried and stirred continuously under heat until the mixtures were dried. The excessive oil in the dried fish floss products was then separated and removed automatically using a spinner machine (Spiner Abon, Indonesia). All experimentations were carried out in triplicate.
Formulation for preparation of little tuna floss products.
Ingredients
Control little tuna floss
Banana blossom addition
12.5%
25%
37.5%
50%
Little tuna meat (g)
1000
870.5
750
620.5
500
Banana blossom (g)
0
120.5
250
370.5
500
Coconut milk (L)
1
1
1
1
1
Onion (g)
40
40
40
40
40
Garlic (g)
10
10
10
10
10
Ginger (g)
10
10
10
10
10
Sugar (g)
10
10
10
10
10
NaCl (g)
80
80
80
80
80
Candlenut (g)
5
5
5
5
5
Brown sugar (g)
100
100
100
100
100
Lemongrass leaves (10 g)
20
20
20
20
20
Lime leaves (8 pieces)
8
8
8
8
8
Preparation of tuna floss incorporated with banana floss.
Nutritional attributeProximate analysis
The proximate analysis of little tuna floss samples was determined according to the method of the Association of Official Analytical Chemists (AOAC, 2000). The Kjeldahl method was used to obtain the crude protein from all dried fish floss samples, while fat content was analyzed using the Soxhlet extractor method. The ash and moisture contents of the samples were examined by the gravimetric method. Carbohydrate content was measured by subtracting total crude protein, fat, ash, and moisture contents from 100%. For energy value (kcal.100g-1), the calculation used the following formula provided by AACC (2000):
Energy value = (4×carbohydrate content) + (4×proteincontent) + (9×lipid content)
Dietary fibre analysis
The dietary fibre analysis used in this study referred to the method of AOAC 985.29 (AOAC, 2000). The little tuna floss samples were dried at 100 °C to constant weight using air-oven (M720, Binder, Germany). Around 0.5 g of all samples were added α-amylase enzyme and kept in accordance with its optimum condition (viz. pH 6; at 100 °C, for 30 min). Afterward, the treated samples were added protease with incubating at 60 °C for 30 min at optimum pH (7.5). The protease-treated samples were then added amyl-glucosidase and incubated at pH 6 for 30 min at optimum temperature. Total dietary fibre of all treated samples was measured by precipitating using ethanol, followed by washing and drying. The obtained residues were then weighed manually.
Amino acids analysis
The amino acids composition was analyzed using ultrapressure liquid chromatography (UPLC) (Waters, US) according to the method of Nollet and Fidel (2015). About 5 mL of hydrochloric acid 6 N was mixed to 0.1 g of samples. The prepared samples were then hydrolyzed at 110 °C for 24 h (M720, Binder, Germany). The hydrolysed samples were transferred to prepared distilled water. After that, the mixtures were filtered with a 0.45 μm filter paper. The 500 μL filtrates were mixed with 40 μL of 2-Amino-4-boronobutanoic acid (ABBA) and 460 μLof double distilled water. Then, 10 μL of the solution was mixed with 70 μL of AccQ Fluorine Borate and 20 μL of fluorine. The homogenized solutions were incubated at 55 °C for 10 min. Finally, the solutions were injected into the UPLC system to calculate the amino acid composition.
Fatty acids analysis
The fatty acid composition of little tuna floss was analysed by PT. Saraswanti Indo Genetech, Bogor Indonesia. Fatty acid methyl esters (FAMEs) from extracted fat of tuna floss samples were prepared by basic transesterification following the official method (AOAC, 2000), using hexane and hydroxide potassium 2N.
FAMEs were analyzed by gas-chromatography (GC) (Agilent Technologies, California, US) described by Aquilani et al., (2018). The GC with a flame ionization detector (FID) was used to analyze the amount of fatty acids in the little tuna floss samples. The GC analysis was conducted using capillary column polyethylene glycol, equivalent variants CP-Wax 52 CB (30m × 0.25mm × 0.25μm film thickness). The carrier gas was nitrogen at a flow rate of 1 mL.min-1. The initial oven temperature was set at 120 °C with an increase of 5 °C per minute until reached 240 °C. The injector and detector temperatures were 260 °C. Individual peaks were measured by comparison of their retention times (RT) with those of standards. Peak areas were calculated, and FAMEs were expressed as the area percentage of total area FAMEs (%).
Heavy metal analysis
Heavy metals analyzed in this study were composed of cadmium (Cd), lead (Pb), arsenic (As), mercury (Hg), and tin (Sn). These microelements were conducted by the method of AOAC (2000) using the Atomic Absorption Spectrophotometer (AAS) (GBC Scientific Equipment, USA).
Histamine determination
The histamine content was carried out by High- Performance Liquid Chromatography (HPLC) (Agilent Technologies, California, US) according to the method from Muscarella et al., (2005). Around 10 g of homogenized little tuna floss was extracted by 20 mL of 5% trichloroacetic acid (TCA). After filtration, 1 mL of the extracted sample was purified with 2 mL of chloroform solution. The mixture was then centrifuged and 20 μL of the supernatant was analyzed by HPLC. The HPLC column used was a Supelcosil LC-ABZ 4.6 × 150 mm, 5 μm film thickness (Supelco, Bellefonte, PA, USA). The decansulphonate phosphate buffer pH 6.9/acetonitrile mixture (85/15) was used as a mobile phase. Isocratic elution was used; the flow rate was 1.2 mL.min-1. The UVDAD was regulated at the absorption wavelength of 210 nm; the injection loop was 20 μL.
Bacterial count
Bacterial evaluations in the tuna floss consist of total bacteria count, total coliform, Escherichia coli, Salmonella sp., and Staphylococcus aureus with the methods developed by Anang et al. (2018) with some modifications. For total bacteria count, serial dilutions of 10 – 1 to 10 – 5 were prepared by diluting around 1 g of the little tuna floss samples into 9 mL of distilled water. Approximately 0.1 mL of aliquots from 10 – 3 to 10 – 5 dilutions were inoculated into Petri dishes containing the Nutrient Agar media. The plates were then incubated at 37 °C for 24 h. After incubation, bacteria colonies formed were counted using the colony counter and recorded as total viable counts. Total coliform was analyzed by dropping 0.1 mL of diluted samples into MacConkey broth with Durham tubes and incubated at 37 °C for 24 h. After incubation, the inoculated tubes were identified as positive total coliform by changing color from purple to yellow, and gas was collected in the Durham tubes. The positive tubes were then transferred into a 5 mL test tube of tryptone solution and incubated at 44 °C for 24 h to test E. coli. Afterward, a drop of Kovacs’ reagent was added into the treated tubes. The suspected E. coli in each tube was shown by a red ring color development, indicating the presence of indole. Salmonella sp., the count was evaluated using pre-enrichment of bovine peptone water, followed by enrichment of Rappaport-Vassiliadis (RV) broth. After enrichment, around 0.1 mL of aliquots were inoculated into Xylose Lysine Deoxycholate (XLD) agar. The suspected Salmonella colonies appear colorless with a black center because of H2S production. For S. aureus count, about 1 g of fish floss samples were added into test tubes containing 9 mL of distilled water and the mixtures were serially diluted from 10 – 1 to 10 – 5. Around 0.1 mL of aliquots (10 – 3 to 10 – 5) were inoculated into plates prepared with Baird Parker Agar media. Afterward, the inoculated samples were incubated at 36 °C for 48 h. After the incubation period, colonies formed were identified with the suspected colonies were round, convex with a diameter of 2 – 3 mm, greyish black color with a clear circle (halo).
Sensory evaluation
Sensory parameters, including appearance, aroma, color, taste, and texture were evaluated using 30 untrained sensory panelists randomly selected among students and this method was adopted from Sęczyk, Świeca and Gawlik-Dziki (2016). The panelists were initially presented with the little tuna floss samples in identical but labeled containers with a three-digit code for their evaluation. Before the sensory session, the tuna floss samples were prepared in triplicate in a randomized permutation. This study used a hedonic test with a 9-point scale to obtain the acceptability score of the little tuna floss products. The likeness scale was arranged in accordance with the above sensory parameters was as follows: 1 = dislike extremely; 2 = dislike very much; 3 = dislike moderately; 4 = dislike slightly; 5 = neither like nor dislike; 6 = like slightly; 7 = like moderately; 8 = like very much; 9 = like extremely.
Statistical analysis
The experimental design applied in this study was a completely randomized design (CRD). All measurements were performed in triplicate. Data were expressed as the mean values ± standard deviation (SD). The differences were determined using a one-way analysis of variance (ANOVA), followed by Duncan’s test. The significant difference was established at p <0.05 using SPSS, Version 27, statistical software program (SPSS Inc., Chicago, Ill., USA).
RESULTS AND DISCUSSIONNutritional attributesProximate composition
The proximate composition of little tuna floss (as a control) and floss samples incorporated with banana blossom with different levels (12.5 – 50%) is presented in Table 2. Results showed a significant (p <0.05) decreased in protein and fat contents of treated tuna floss samples with the increase in the percentage of banana blossom.
Proximate composition (g.100g-1), energy value (Kcal) and histamine content (mg.kg-1) of the control sample and added little tuna floss with banana blossom.
Parameter
Control little tuna floss
Banana blossom addition
SNI*
12.5%
25%
37.5%
50%
Moisture
16.03±0.01a
16.45±0.38a
16.74±0.27a
17.23±0.16b
17.39±0.06b
<7
Crude protein
43.08±1.14d
30.27±1.41c
29.51±1.24b
28.68±0.30a
28.13±0.16a
>15
Crude fat
18.70±0.31c
18.02±0.07c
16.62±0.06b
15.20±0.18a
14.79±0.08a
<30
Ash
4.12±0.03a
4.45±0.11a
4.52±0.06a
4.78±0.22a
5.68±0.13b
>7
Crude fibre
1.8±0.42a
2.6±0.21a
2.8±0.50b
3.1±0.43b
3.5±0.59c
<1
Carbohydrate
15.07±0.58a
27.81±1.04b
29.61±0.43c
31.11±0.27d
31.01±0.20d
-
Energy
271.66±1.01a
303.69±1.84d
297.53±0.52c
289.92±0.25b
285.28±0.14b
-
Histamine
319.48±121e
144.16±1.18d
110.56±1.68c
85.09±1.68b
52.25±1.35a
<200**
Note: Values are given as mean ± standard deviation from triplicate determinations (n = 3). Different superscript letters in the same row indicate significant differences (p <0.05). * SNI 01-3707-1995 is used for fish floss specification. ** Histamine limit referred to FAO/WHO (2012).
However, the protein and fat contents of all little tuna floss samples agreed with the Indonesian National Standard (SNI) values and were even much better than a limit recommended by the standard (SNI 7988, 2009) and for protein content, all treated samples were in accordance with other fish flosses (Huda et al., 2012;Wijaya et al., 2016;Romadhon, Amalia and Anggo, 2019;Fahmi and Purnamayati, 2020).
The higher protein indicates the developed tuna floss samples in this study are potentially used as a filler for low-protein food products, such as glutinous rice rolls, bread, and pastel products. The most significant effect of the formulation is the improvement in crude fibre content of the control little tuna flosses, which increased by 44.44 – 94.44% with increasing the percentage of banana blossom from 12.5 – 50%. The increase in crude fibre content of the control tuna floss with the increase in the percentage of banana blossom was similar to the results obtained for floss incorporated with other fibre sources (Bujang et al., 2016;Candra and Tunoq, 2018;Poonsri et al., 2019) and its fibre content was higher than the values determined by SNI and other fish floss products. The higher fibre content is an indication of the better quality of the products with the potential health benefits as above mentioned. The contents of ash, carbohydrate, and moisture increased significantly (p <0.05) with incorporating banana blossom in the little tuna flosses. In terms of ash content, the formulated floss samples agreed with the SNI requirements (>7) for the standardized fish floss product. Moreover, the energy values of treated samples (285.28 – 303.69 kcal) were higher than the control (271.66 kcal); but these values were slightly lower than the dietary requirements recommended by the FAO/WHO/UNU (2007) for children and young adult foods. The high energy level and protein content in the tuna floss added with banana blossom indicates its suitability as a supporting diet for children and young adults.
The effect of the addition of banana blossom into a tuna floss formula on histamine content is also depicted in Table 2. Interestingly, the histamine contents of treated tuna floss samples decreased significantly (p <0.05) with increasing the level of banana blossom. Compared to the control sample (319.48 mg.kg-1), around 2-fold (144.16 mg.kg-1) to 6-fold (52.25 mg.kg-1) of histamine content decreased with the increment of at around 12.5% to 50% into tuna floss samples, respectively. The reduction of histamine in the treated tuna floss might be due to the ratio of banana blossom and tuna as main materials in the development of little tuna floss products, in which a higher banana blossom was added, a lower content of histamine found in the tuna floss samples. These results were lower than found in the tuna dumpling (1608 mg.kg-1) studied by Chen et al. (2008), while the level of histamine in the present study showed a higher compared to the study from Peivasteh-Roudsari et al. (2020) in the canned tuna (34.46 mg.kg-1). Nonetheless, the histamine concentration in all treated flosses was below the 200 mg.kg-1 allowable limitation suggested by the FAO/WHO (2012), which would not cause an adverse effect. This histamine limit is also under the European Union Regulation (EC) No 2073/2005 (EC, 2005) for fishery products. It can be inferred that all treated tuna floss samples are safe as a food product.
Amino acids composition
Table 3. presents the profile of amino acids found in the control sample and the tuna floss incorporated with different concentrations of banana blossom. Results exhibited that glutamic acid (Glu) was the most dominant amino acid in all little tuna flosses. However, 15-amino acid analyzed in the control sample was a higher amount of total amino acids both essential and non-essential amino acids than found in the treated tuna floss products. This is due to the lower amino acids content observed in banana blossom incorporated with tuna floss samples (Table 3). In terms of essential amino acids, the control and treated samples showed higher values compared to the nonessential amino acids. Furthermore, the content of essential amino acids, including arginine, histidine, isoleucine. leucine, lysine, phenylalanine, threonine, and valine, found in the treated tuna floss samples did not meet the standard recommended by Food and Agriculture Organisation/World Health Organisation/United Nations University (FAO/WHO/UNU) for children and adult humans (FAO/WHO/UNU, 2007).
Amino acids composition (mg.g-1 protein) of control sample and tuna floss incorporated with banana blossom.
Amino acids
Banana blossom
Control tuna floss
Banana blossom addition
RDA*
12.5%
25%
37.5%
50%
Ala
0.88±0.00
12.60±0.04e
10.43±0.03d
6.92±0.03c
6.43±0.01b
6.27±0.02a
-
Arg
1.36±0.01
32.81±0.05e
26.73±0.07d
16.05±0.04c
14.68±0.29b
11.13±0.03a
20
Asp
1.58±0.00
34.90±0.13e
28.94±0.07d
20.54±0.10c
19.65±0.16b
17.71±0.01a
-
Glu
2.08±0.02
55.88±0.17e
46.82±0.12d
32.25±0.08c
30.86±0.05b
28.80±0.06a
-
Gly
1.05±0.01
24.43±0.05e
20.34±0.03d
12.76±0.01c
11.82±0.23b
10.17±0.02a
-
His
0.62±0.01
25.31±0.10e
19.71±0.09d
13.22±0.06c
10.43±0.14b
8.55±0.02a
15-16
Ile
0.94±0.01
24.29±0.08e
19.81±0.05d
11.93±0.04c
11.55±0.11b
10.46±0.02a
30-31
Leu
1.11±0.01
40.84±0.15e
33.57±0.08d
20.25±0.08c
19.23±0.03b
18.08±0.02a
59-61
Lys
0.78±0.01
32.12±0.10e
26.41±0.08d
18.57±0.06c
17.96±0.01b
17.26±0.03a
45-48
Phe
0.76±0.01
26.69±0.15e
21.02±0.03d
13.63±0.00c
11.09±0.57b
8.63±0.03a
38
Pro
0.90±0.01
15.14±0.05d
12.76±0.04c
8.11±0.03b
7.40±0.02a
7.38±0.02a
-
Ser
1.40±0.01
24.42±0.08e
19.47±0.06d
11.53±0.03c
10.62±0.08b
9.37±0.02a
-
Thr
0.86±0.01
28.99±0.09e
23.60±0.09d
14.09±0.06c
13.63±0.06b
10.80±0.03a
23-25
Tyr
0.67±0.00
16.27±0.07e
13.67±0.03d
8.25±0.04c
6.46±0.03b
5.24±0.03a
-
Val
0.94±0.01
27.32±0.09d
22.24±0.09c
13.98±0.02b
12.84±0.06a
12.67±0.01a
39-40
TEAA**
8.05
254.56e
206.76d
129.97c
118.35b
102.82a
-
TNEAA***
7.89
167.37e
138.76d
92.12c
86.27b
79.71a
-
Note: Values are given as mean ± standard deviation from triplicate determinations (n = 3). Different letters in the same row indicate significant differences (p <0.05). * RDA: Recommended dietary allowance for children and adult humans by FAO/WHO/UNU (2007). ** TEAA: total essential amino acids (arginine, histidine, isoleucine. leucine, lysine, phenylalanine, threonine and valine). *** TNEAA: total non-essential amino acids (aspartic acid, glutamic acid, serine, tyrosine, glycine and alanine).
Nevertheless, fish floss is a non-staple food product like bread, noodle, and rice, which is routinely consumed in large quantities to provide adequate energy, but it is generally combined with other staple foods to enhance nutritional values.
Fatty acids composition
The effect of the formulation of tuna floss with banana blossom on the fatty acid composition is presented in Table 4. The content of fatty acids in the tuna floss samples, both control and treated samples with different levels of banana blossom, varied considerably (p <0.05). The most abundant fatty acids in all treated floss samples were oleic acid (C18:1) (34.03 – 40.63%), followed by palmitic acid (C16:0) (27.19 – 31.84%) and linoleic acid (C18:2) (11.20 – 13.30%), but these fatty acids were slightly lower compared to the control sample’s fatty acids. It might be due to a low-fatty acids content observed in the banana blossom (0.39 – 1.28%). In addition, all tuna floss incorporated with banana blossom had lower content of saturated (33.84 – 41.44%), monounsaturated (34.37 – 41.00%), and polyunsaturated (13.72 – 16.29%) fatty acids than those contained in the control. However, among the identified fatty acids, omega-3 (n-3) and omega-6 (n-6) fatty acids seem to be the most important, due to their multiple biological roles, such as reducing oxidative stress, influencing the inflammatory cascade, presenting neuroprotection, and cardiovascular protection. The total n-6 and n-3 in the treated samples ranged at around 11.28 – 13.40% and 2.44 – 3.23% respectively, with an n-6/n-3 ratio of about 3.89 – 4.62%. These values within the range (1 – 5) of omega-6 and omega-3 ratio per day recommended by some food experts that should be consumed to prevent undesirable diseases related to the lack of essential fatty acids intake (EFSA, 2010). Moreover, amongst n-3 fatty acids, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are required essentially by the human body and approximately 0.2-2.0 g/day recommended by most health organizations (Desai et al., 2018), and all formulated tuna floss samples met the requirements.
Fatty acids profile (%) of control tuna floss and treated tuna floss with addition of banana blossom.
Fatty acids
Banana blossom
Control tuna floss
Banana blossom addition
12.5%
25%
37.5%
50%
C6:0
n.d.
0.04 ±0.000b
0.03 ±0.001a
0.04 ±0.000b
n.d.
n.d.
C8:0
n.d.
0.38 ±0.007c
0.37 ±0.006c
0.44 ±0.009d
0.23 ±0.001b
0.17 ±0.000a
C10:0
n.d.
0.25 ±0.002c
0.26 ±0.003c
0.33 ±0.006d
0.16 ±0.002b
0.13 ±0.001a
C12:0
n.d.
2.15 ±0.051c
n.d.
2.68 ±0.063d
1.35 ±0.005b
1.14 ±0.000a
C14:0
0.02 ±0.000
1.71 ±0.014a
1.76 ±0.041a
1.85 ±0.012b
1.20 ±0.012b
1.25 ±0.001c
C15:0
n.d.
0.05 ±0.000b
0.04 ±0.000a
0.04 ±0.000a
0.04 ±0.001a
0.03 ±0.000a
C16:0
0.92 ±0.010
32.52 ±0.056c
31.84 ±0.433c
28.83 ±0.115b
27.47 ±0.224a
27.19 ±0.016a
C16:1
0.02 ±0.000
0.19 ±0.000
0.17 ±0.004
0.16 ±0.001
0.16 ±0.001
0.15 ±0.001
C17:0
n.d.
0.12 ±0.000
0.10 ±0.001
0.10 ±0.000
0.10 ±0.001
0.09 ±0.000
C17:1
n.d.
0.04 ±0.001
0.03 ±0.000
0.03 ±0.001
0.02 ±0.000
0.03 ±0.000
C18:0
0.12 ±0.000
4.53 ±0.004e
4.31 ±0.061d
3.93 ±0.004c
3.68 ±0.024b
3.47 ±0.002a
C18:1n9
0.39 ±0.001
41.99 ±0.020e
40.63 ±0.323d
35.86 ±0.001c
34.97 ±0.139b
34.03 ±0.012a
C18:2n6
1.28 ±0.000
14.02 ±0.008d
13.30 ±0.106c
12.42 ±0.001b
11.25 ±0.042a
11.20 ±0.000a
C18:3n3
0.36 ±0.000
2.45 ±0.000e
2.17 ±0.023c
2.28 ±0.002d
1.68 ±0.008a
1.79 ±0.000b
C20:0
0.24 ±0.003
0.38 ±0.006d
0.35 ±0.007c
0.30 ±0.002b
0.29 ±0.007b
0.27 ±0.001a
C20:1
0.14 ±0.000
n.d.
0.17 ±0.004
0.16 ±0.000
0.16 ±0.002
n.d.
C20:4n6
n.d.
0.11 ±0.001c
0.10 ±0.002b
0.12 ±0.002d
0.12 ±0.003d
0.08 ±0.001a
C20:5n3
n.d.
0.20 ±0.003c
0.18 ±0.004a
0.19 ±0.002b
0.21 ±0.003c
0.17 ±0.000a
C21:1
n.d.
0.18 ±0.001
n.d.
n.d.
n.d.
n.d.
C22:6n3
n.d.
0.64 ±0.012c
0.55 ±0.008b
0.76 ±0.006d
0.67 ±0.016c
0.48 ±0.000a
C24:0
0.02 ±0.000
0.13 ±0.002
0.10 ±0.001
0.10 ±0.002
0.12 ±0.000
0.10 ±0.001
Σ SFA
1.38 ±0.005
42.25 ±0.031e
41.44 ±0.443d
38.63 ±0.030c
34.65 ±0.197b
33.84 ±0.027a
Σ MUFA
0.62 ±0.001
42.40 ±0.019e
41.00 ±0.323d
36.21 ±0.001c
35.31 ±0.139b
34.37 ±0.012a
Σ PUFA
1.64 ±0.000
17.42 ±0.009d
16.29 ±0.143c
15.77 ±0.008b
13.93 ±0.073a
13.72 ±0.001a
Σn6
1.28 ±0.000
14.13 ±0.004d
13.40 ±0.004c
12.54 ±0.001b
11.37 ±0.004a
11.28 ±0.001a
Σn3
0.36 ±0.000
3.29 ±0.007e
2.90 ±0.011c
3.23 ±0.005d
2.56 ±0.009b
2.44 ±0.000a
PUFA/SFA
6.54 ±0.025
2.27 ±0.001b
2.16 ±0.042a
2.25 ±0.003b
2.21 ±0.024ab
2.23 ±0.002ab
EPA+DHA
n.d.
0.84 ±0.015c
0.73 ±0.012b
0.95 ±0.009d
0.88 ±0.019c
0.65 ±0.000a
n6/n3
3.56
4.29
4.62
3.89
4.44
4.62
Note: Values are given as mean ± standard deviation from triplicate determinations (n = 3). Different superscript letters in the same row indicate significant differences (p <0.05).
Heavy metal content
Heavy metals analyzed in this study are cadmium (Cd), lead (Pb), arsenic (As), mercury (Hg), and tin (Sn). These are classified as toxic metals and endanger human health if the total content of the metals exceeds the recommended exposure limits (Sajib et al., 2014;Lukáčová et al., 2014;Timoracká, Vollmannová and Ismael, 2017). As presented in Table 5, the formulation of tuna floss with different levels (12.5 – 50%) of banana blossom had a wide range of metal element compositions such as Cd (<16×10 – 4 mg.kg-1), Pb (<20×10 – 4 mg.kg-1), As (8×10 – 4 mg.kg-1), Hg (1×10 – 4 mg.kg-1) and Sn (36.15 – 37.67 mg.kg-1).
Heavy metals content (mg.kg-1) of control and added tuna floss samples with banana blossom.
Elements
Control little tuna floss
Banana blossom addition
SNI*
12.5%
25%
37.5%
50%
Cd
<0.0016
<0.0016
<0.0016
<0.0016
<0.0016
<0.50
Pb
<0.0020
<0.0020
<0.0020
<0.0020
<0.0020
<2.00
As
<0.0008
<0.0008
<0.0008
<0.0008
<0.0008
<1.00
Hg
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
<0.05
Sn
27.53±0.41a
36.15±0.120b
34.21±0.10b
37.67±0.20b
37.3±0.20b
<40.00
Note: Values are given as mean ± standard deviation from triplicate determinations (n = 3). Different superscript letters in the same row indicate significant differences (p <0.05). *SNI 01- 3707-1995 is used for fish floss specification.
Compared to the control treatment, all identified elements were relatively the same in the contents except amount of Sn, which the content of Sn increased significantly (p <0.05). However, all heavy metals contained in the treated samples agreed with the Indonesian National Standard for fish floss specification (SNI 01-3707, 1995). It indicates that the tuna floss incorporated with a banana blossom is potentially safe for human beings.
Bacterial test
Table 6. shows the bacterial counts for all tuna floss samples evaluated with different tests, including total count, Salmonella sp., E. coli, S. aureus, and total coliform. For total bacterial count, a non-selective media was used, and the results presented that no significant difference (p >0.05) was exhibited in the total bacterial count and slightly higher compared to scad fish (Decapterus sp.) floss (Kasmiati et al., 2020). However, all treated samples did not exceed the maximum permissible limits of microbial contamination in food products regulated by the Indonesian National Standard (SNI 7988, 2009). The identification of Salmonella sp., E. coli, S. aureus in the tuna floss samples using specific media showed no growth (negative) after incubation and these results agreed with the requirement for microbial contamination in the dried fish product (SNI 7988, 2009). Furthermore, all treated samples, both augmented and unaugmented with banana blossom, showed a similar count (<3 MPN.g-1) to the total coliform. These results revealed that the total coliform in all tested samples were within the acceptable microbial quality of the Indonesia National Standard. From those microbial evaluations, the tuna floss product used in the present study agrees with the SNI 7988: 2009.
Bacterial count of control tuna floss and tuna flosses added with banana blossom.
Bacterial test
Control tuna floss
Banana blossom addition
SNI*
12.5%
25%
37.5%
50%
Total count (cfu.g-1)
3.4 × 104
1.6 × 104
3.3 × 104
5.2 × 104
1.1 × 104
<1× 105
Salmonellasp. (cfu.g-1)
negative
negative
negative
negative
negative
negative
E. coli (cfu.g-1)
negative
negative
negative
negative
negative
< 3
S. aureus (cfu.g-1)
negative
negative
negative
negative
negative
<1×102
Total coliform (MPN.g-1)
<3
<3
<3
<3
<3
<10
Note: *SNI 7988: 2009: maximum permissible limits of microbial contamination in food products.
Sensory attribute
Sensory evaluation is a suitable tool for developing food products with assessing consumer’s acceptance (Anang et al., 2018;Fiorentini, Kinchla and Nolden, 2020; Witczak, Jaworska and Witczak, 2020).
Assessment of sensory attributes (appearance, aroma, color, taste, texture, and overall quality) of tested tuna floss by the panelist is depicted in Figure 2. Generally, the addition of banana blossom at the level from 12.5% to 50% to tuna flosses had a significant influence (p <0.05) on its sensory characteristics and consequently on consumer acceptance. Amongst these formulations, the 37.5% added floss showed the highest scores in almost assessed attributes compared to other treatments and control samples.
Physical appearance and sensory attributes of control tuna floss and tuna floss incorporated with different levels (12.5 – 50%) of banana blossom. The same letters denote the lack of statistically significant differences between the results at p <0.05 (n = 30).
In addition, the mean score of overall acceptability in the tested floss samples was approximately 7.11, which implies that the treatment of tuna flosses is preferred moderately among the consumers. This acceptability value agreed with the requirement recommended by the Indonesian National Standard (5) (SNI 01-3707, 1995), and even much higher than the score accepted by the SNI. Also, these results are in accordance with the studies from Candra and Tunoq (2018), Puspita, Kartikaningsih and Dayuti (2019) and Novidiyanto et al., (2020) used the addition of banana flowers into snakehead (Channa striata), Indian scad (Decapterusrusselli) and chicken floss products, respectively.
CONCLUSION
Taken together, the addition of tuna floss with different levels (12.5% – 50%) of banana blossom reduced histamine level and increased the dietary fibre content in the little tuna floss samples. In addition, microbial and toxic elements were permissible limits regulated by the Indonesian National Standard (SNI) in all the treated tuna flosses. Our results suggested that 37.5% of banana blossom incorporated into the tuna flosses was selected as an appropriate formula due to its attributes, such as being more acceptable for overall sensory evaluation, high contents of EPA+DHA, and dietary fibre, as well as a low histamine content. However, our further research concerns the nutritional value of the selected tuna floss sample, especially on the enrichment of essential amino acids composition through fortification strategy. Also, functional properties like antioxidant and antimicrobial activities will be further studied.
Acknowledgments:
The authors are thankful for the financial support provided by the Universitas Brawijaya, Indonesia through the Research Grant for Professor and Doctor 2020, grant number 35, 2020. Also, we are grateful for the payment of article processing charge (APC) of the Universiti Malaysia Sabah, for joining publication.
Conflict of Interest:
The authors declare no conflict of interest.
Ethical Statement:
This article does not contain any studies that would require an ethical statement.
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