Potravinarstvo Slovak Journal of Food Sciences
Volume 15 846 2021



Potravinarstvo Slovak Journal of Food Sciences
vol. 15, 2021, p. 846-857
https://doi.org/10.5219/1657
Received: 25 June 2021. Accepted: 26 August 2021.
Available online: 28 September 2021 at www.potravinarstvo.com
© 2021 Potravinarstvo Slovak Journal of Food Sciences, License: CC BY 4.0
ISSN 1337-0960 (online)

THE NUTRITIONAL VALUE, BACTERIAL COUNT AND SENSORY
ATTRIBUTES OF LITTLE TUNA ( EUTHYNNUS AFFINIS) FLOSS
INCORPORATED WITH THE BANANA BLOSSOM

Hartati Kartikaningsih, Yahya, Yuniar Tri Hartita, Abdul Aziz Jaziri, Wahidu Zzaman,
Rovina Kobun, Nurul Huda

ABSTRACT
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, alpha-
tocopherol, 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

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 847 2021
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 METHODOLOGY
Samples
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 experiment
Sample 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

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 848 2021
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.

Nutritional attribute
Proximate 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 × protein
content) + (9 × lipid content) (1)
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 ultra-
pressure 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.

Table 1 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

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 849 2021



Figure 1 Preparation of tuna floss incorporated with banana floss.

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 850 2021
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 DISCUSSION
Nutritional attributes
Proximate 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.

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 851 2021
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 non-
essential 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
Table 2 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±0.01
a
16.45±0.38
a
16.74±0.27
a
17.23±0.16
b
17.39±0.06
b
<7
Crude protein 43.08±1.14
d
30.27±1.41
c
29.51±1.24
b
28.68±0.30
a
28.13±0.16
a
>15
Crude fat 18.70±0.31
c
18.02±0.07
c
16.62±0.06
b
15.20±0.18
a
14.79±0.08
a
<30
Ash 4.12±0.03
a
4.45±0.11
a
4.52±0.06
a
4.78±0.22
a
5.68±0.13
b
>7
Crude fibre 1.8±0.42
a
2.6±0.21
a
2.8±0.50
b
3.1±0.43
b
3.5±0.59
c
<1
Carbohydrate 15.07±0.58
a
27.81±1.04
b
29.61±0.43
c
31.11±0.27
d
31.01±0.20
d
-
Energy 271.66±1.01
a
303.69±1.84
d
297.53±0.52
c
289.92±0.25
b
285.28±0.14
b
-
Histamine 319.48±121
e
144.16±1.18
d
110.56±1.68
c
85.09±1.68
b
52.25±1.35
a
<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).

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 852 2021
University (FAO/WHO/UNU) for children and adult
humans (FAO/WHO/UNU, 2007).

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.

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
).


Table 3 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±0.04
e
10.43±0.03
d
6.92±0.03
c
6.43±0.01
b
6.27±0.02
a
-
Arg 1.36±0.01 32.81±0.05
e
26.73±0.07
d
16.05±0.04
c
14.68±0.29
b
11.13±0.03
a
20
Asp 1.58±0.00 34.90±0.13
e
28.94±0.07
d
20.54±0.10
c
19.65±0.16
b
17.71±0.01
a
-
Glu 2.08±0.02 55.88±0.17
e
46.82±0.12
d
32.25±0.08
c
30.86±0.05
b
28.80±0.06
a
-
Gly 1.05±0.01 24.43±0.05
e
20.34±0.03
d
12.76±0.01
c
11.82±0.23
b
10.17±0.02
a
-
His 0.62±0.01 25.31±0.10
e
19.71±0.09
d
13.22±0.06
c
10.43±0.14
b
8.55±0.02
a
15-16
Ile 0.94±0.01 24.29±0.08
e
19.81±0.05
d
11.93±0.04
c
11.55±0.11
b
10.46±0.02
a
30-31
Leu 1.11±0.01 40.84±0.15
e
33.57±0.08
d
20.25±0.08
c
19.23±0.03
b
18.08±0.02
a
59-61
Lys 0.78±0.01 32.12±0.10
e
26.41±0.08
d
18.57±0.06
c
17.96±0.01
b
17.26±0.03
a
45-48
Phe 0.76±0.01 26.69±0.15
e
21.02±0.03
d
13.63±0.00
c
11.09±0.57
b
8.63±0.03
a
38
Pro 0.90±0.01 15.14±0.05
d
12.76±0.04
c
8.11±0.03
b
7.40±0.02
a
7.38±0.02
a
-
Ser 1.40±0.01 24.42±0.08
e
19.47±0.06
d
11.53±0.03
c
10.62±0.08
b
9.37±0.02
a
-
Thr 0.86±0.01 28.99±0.09
e
23.60±0.09
d
14.09±0.06
c
13.63±0.06
b
10.80±0.03
a
23-25
Tyr 0.67±0.00 16.27±0.07
e
13.67±0.03
d
8.25±0.04
c
6.46±0.03
b
5.24±0.03
a
-
Val 0.94±0.01 27.32±0.09
d
22.24±0.09
c
13.98±0.02
b
12.84±0.06
a
12.67±0.01
a
39-40
TEAA*
*
8.05 254.56
e
206.76
d
129.97
c
118.35
b
102.82
a
-
TNEAA
***
7.89 167.37
e
138.76
d
92.12
c
86.27
b
79.71
a
-
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).

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 853 2021

Table 4 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 ±0.000
b
0.03 ±0.001
a
0.04 ±0.000
b
n.d. n.d.
C8:0 n.d. 0.38 ±0.007
c
0.37 ±0.006
c
0.44 ±0.009
d
0.23 ±0.001
b
0.17 ±0.000
a

C10:0 n.d. 0.25 ±0.002
c
0.26 ±0.003
c
0.33 ±0.006
d
0.16 ±0.002
b
0.13 ±0.001
a

C12:0 n.d. 2.15 ±0.051
c
n.d. 2.68 ±0.063
d
1.35 ±0.005
b
1.14 ±0.000
a

C14:0 0.02 ±0.000 1.71 ±0.014
a
1.76 ±0.041
a
1.85 ±0.012
b
1.20 ±0.012
b
1.25 ±0.001
c

C15:0 n.d. 0.05 ±0.000
b
0.04 ±0.000
a
0.04 ±0.000
a
0.04 ±0.001
a
0.03 ±0.000
a

C16:0 0.92 ±0.010 32.52 ±0.056
c
31.84 ±0.433
c
28.83 ±0.115
b
27.47 ±0.224
a
27.19 ±0.016
a

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 ±0.004
e
4.31 ±0.061
d
3.93 ±0.004
c
3.68 ±0.024
b
3.47 ±0.002
a

C18:1n9 0.39 ±0.001 41.99 ±0.020
e
40.63 ±0.323
d
35.86 ±0.001
c
34.97 ±0.139
b
34.03 ±0.012
a

C18:2n6 1.28 ±0.000 14.02 ±0.008
d
13.30 ±0.106
c
12.42 ±0.001
b
11.25 ±0.042
a
11.20 ±0.000
a

C18:3n3 0.36 ±0.000 2.45 ±0.000
e
2.17 ±0.023
c
2.28 ±0.002
d
1.68 ±0.008
a
1.79 ±0.000
b

C20:0 0.24 ±0.003 0.38 ±0.006
d
0.35 ±0.007
c
0.30 ±0.002
b
0.29 ±0.007
b
0.27 ±0.001
a

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 ±0.001
c
0.10 ±0.002
b
0.12 ±0.002
d
0.12 ±0.003
d
0.08 ±0.001
a

C20:5n3 n.d. 0.20 ±0.003
c
0.18 ±0.004
a
0.19 ±0.002
b
0.21 ±0.003
c
0.17 ±0.000
a

C21:1 n.d. 0.18 ±0.001 n.d. n.d. n.d. n.d.
C22:6n3 n.d. 0.64 ±0.012
c
0.55 ±0.008
b
0.76 ±0.006
d
0.67 ±0.016
c
0.48 ±0.000
a

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
∑ SFA 1.38 ±0.005 42.25 ±0.031
e
41.44 ±0.443
d
38.63 ±0.030
c
34.65 ±0.197
b
33.84 ±0.027
a

∑ MUFA 0.62 ±0.001 42.40 ±0.019
e
41.00 ±0.323
d
36.21 ±0.001
c
35.31 ±0.139
b
34.37 ±0.012
a

∑ PUFA 1.64 ±0.000 17.42 ±0.009
d
16.29 ±0.143
c
15.77 ±0.008
b
13.93 ±0.073
a
13.72 ±0.001
a

∑n6 1.28 ±0.000 14.13 ±0.004
d
13.40 ±0.004
c
12.54 ±0.001
b
11.37 ±0.004
a
11.28 ±0.001
a

∑n3 0.36 ±0.000 3.29 ±0.007
e
2.90 ±0.011
c
3.23 ±0.005
d
2.56 ±0.009
b
2.44 ±0.000
a

PUFA/SFA 6.54 ±0.025 2.27 ±0.001
b
2.16 ±0.042
a
2.25 ±0.003
b
2.21 ±0.024
ab
2.23 ±0.002
ab

EPA+DHA n.d. 0.84 ±0.015
c
0.73 ±0.012
b
0.95 ±0.009
d
0.88 ±0.019
c
0.65 ±0.000
a

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).


Table 5 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±0.41
a
36.15±0.120
b
34.21±0.10
b
37.67±0.20
b
37.3±0.20
b
<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.

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 854 2021
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 un-
augmented 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.
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).
Table 6 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 × 10
4
1.6 × 10
4
3.3 × 10
4
5.2 × 10
4
1.1 × 10
4
<1× 10
5

Salmonella sp. (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 <1×10
2

Total coliform (MPN.g
-1
) <3 <3 <3 <3 <3 <10
Note: *SNI 7988: 2009: maximum permissible limits of microbial contamination in food products.


Figure 2 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).

Potravinarstvo Slovak Journal of Food Sciences
Volume 15 855 2021
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.
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.

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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.

Contact Address:
Hartati Kartikaningsih, Department of Fishery Product
Technology and Bioseafood Research Group, Faculty of
Fisheries and Marine Science, Universitas Brawijaya, Jl.
Veteran 65145, Malang, Indonesia, Tel.: +62341551611,
E-mail: [email protected]
ORCID: https://orcid.org/0000-0002-5124-1512
Yahya, Department of Fishery Product Technology and
Bioseafood Research Group, Faculty of Fisheries and
Marine Science, Universitas Brawijaya, Jl. Veteran 65145,
Malang, Indonesia, Tel.: +62341551611,
E-mail: [email protected]
ORCID: -
Yuniar Tri Hartita, Department of Fishery Product
Technology, Faculty of Fisheries and Marine Science,
Universitas Brawijaya, Jl. Veteran 65145, Malang,
Indonesia, Tel.: +62341551611,
E-mail: [email protected]
ORCID: https://orcid.org/0000-0002-6349-6774
Abdul Aziz Jaziri, Department of Fishery Product
Technology and Bioseafood Research Group, Faculty of
Fisheries and Marine Science, Universitas Brawijaya, Jl.
Veteran 65145, Malang, Indonesia, Tel.: +62341551611,
E-mail: [email protected]
ORCID: https://orcid.org/0000-0001-7121-4055
Wahidu Zzaman, Department of Food Engineering and
Tea Technology, Shahjalal University of Science and
Technology, Sylhet-3114, Bangladesh,
E-mail: [email protected] / [email protected]
ORCID: https://orcid.org/0000-0003-1513-7301
Rovina Kobun, Faculty of Food and Nutrition, Universiti
Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah,
Malaysia, Tel.: +6088320000,
E-mail: [email protected]
ORCID:https://orcid.org/0000-0002-4985-9145
*Nurul Huda, Faculty of Food and Nutrition, Universiti
Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah,
Malaysia, Tel.: +6088320000; Department of Food
Science and Technology, Faculty of Agriculture,
Universitas Sebelas Maret, Jl. Ir. Sutami, Surakarta,
57126, Central Java, Indonesia,
E-mail: [email protected]
ORCID: https://orcid.org/0000-0001-9867-6401

Corresponding author: *