acceptable and popular fish species ... Indonesia. Consequently the purpose of this research was to

International Journal of Bioscience, Biochemistry and Bioinformatics, Vol. 4, No. 2, March 2014
Quality Characteristics of Smoked Skipjack
(Katsuwonuspelamis) Using Different Liquid Smoke
Fronthea Swastawati, Y. S. Darmanto, L. Sya’rani, K. Rahayu Kuswanto, and K. D. Anthony Taylor

acceptable and popular fish species to be smoked in
Indonesia. Consequently the purpose of this research was to
determinate the effects of two liquid smoke flavourings on
the quality variables of smoked skipjack.
Abstract—The application of liquid smoke to improve
quality of smoked skipjack (Katsuwonuspelamis) was studied in
this research. Two different liquid smoke i.e. paddy chaff (PLS)
and coconut shell liquid smoke (CLS)were applied to smoked
fish. Both liquid smoke (PLS and CLS) were determined their
chemical compound such as phenol; pH; and Benzo (α) pyrene,
while the smoked skipjack were analysed in terms of sensory,
microbiological and chemical aspect. The result showed that
total phenolic derivatives found in PLS were 25,1%, lower
than that of CLS of 26,31%; The pH value of PLS was 3,2;
slightly higher than the CLS of 2,5; Benzo (α) pyrene was not
detected in both PLS and CLS; but were found in smoked
skipjack 9.55 and 8.72 ppm respectively. Sensory values of
smoked fish produced by PLS were 7.7 and CLS was 7.8.
Moisture content of the PLS smoked fish was 54.02% lower
than CLS smoked fish that was 54.34%; Examination of t-test
Independent Sample to the moisture content showed no
significantly different (P>0.05); TVC on PLS skipjack was 153
cfu/g very significantly higher (P<0.01) than smoked skipjack
using CLS that was 76 cfu/g.
II. MATERIALS AND METHODS
A. Production of Liquid Smoke Flavouring
The PLS and CLS were collected in around Semarang
City. They were dried for several hours prior to processing.
The pyrolysis process was started with smoke generator
controlled heating. The temperature was measured with a
thermostat positioned in the centre of the reactor; the
maximum temperature reached was 450ºC. The production
was stopped when the smoke materials was totally
pyrolysed.
The liquid smoke resulting from the process was filtered
through a paper filter and collected in some bottles.
Index Terms—Skipjack, paddy chaff (PLS) and coconut
shell (CLS) liquid smoke, quality.
B. Smoked Cured Skipjack Processing
The experimental of smoked skipjack were prepared in
Fish Processing Product Technology Laboratory following
the procedure of Indonesian National Standard/SNI 2006. A
total of 20 kg of skipjack were used in this experiment, and
the fish were collected from Hygienic Fish Market in
Semarang.
In the processing, the fish were eviscerated, cut, and
washed thoroughly, then divided into two groups. One
group was subjected to PLS while the other was treated by
CLS. The fish were dipped into 3% salt and 5% liquid
smoke solution for 15 minutes. Then the cured fish were
placed into an oven and cooked at 80ºC for ±3 hours.
I. INTRODUCTION
Smoking is one of the old food preservation techniques. It
is being more popular nowadays by the use of liquid smoke.
The utilization of liquid smoke offers some advantages
compared to traditional methods. Liquid smokes are easily
to applied, the concentration of liquid smoke can be
controlled, can result in more uniform products, be less
taxing on the environment and many smoke flavourings are
also free of harmful compounds such as polycyclic aromatic
hydrocarbons [1].
Suitable smoke condensates for smoking of particular fish
species could be evaluated by sensory value, as well as
microbiological, chemical and safety point of view. In
related to consumer preferences, it is found that not all
consumers like the same kind of products. For example
some people require a strong smoke odour and flavour,
others want a specific wood or smoke material [2], [3].
Possible materials for use in the production of liquid smoke
in Indonesia are paddy chaff and coconut shell, due to their
chemical composition [4]. Skipjack is one of the most
C. Phenol Analysis
Samples were homogenized in destilled water, placed into
iodide tubes 0,1 N bromine solutions and 5 ml of
concentrated HCl, added and left it for 30 minutes. After
that, 5 mL of 20% KI solutions and 3 mL of 5 % Amylop
indicator were added, and then titrated with Na-Tiosulfat 0,1
N sodium thiosulphate until the blue colour disappeared [5].
D. Determination of pH
20 mL of liquid smoke samples were put into a beaker
glass. Then the pH meter sensor was put into the liquid
smoke sample and the pH value was recorded when the
reading was constant [6]
Manuscript received August 14, 2013; revised November 24, 2013.
Fronthea Swastawati, Y. S. Darmanto, and L. Sya’rani are with
Department of Fisheries, Faculty of Fisheries and Marine Science,
Diponegoro University, Jl. Prof Soedarto, SH, Kampus UNDIP Tembalang,
Semarang (e-mail: fronthea_thp@ undip.ac.id).
K. Rahayu Kuswanto is with the Department of Food Technology from
GadjahMada University (e-mail: kapti_rk@yahoo.com).
K. D. Anthony Taylor is with the Department of Food Science and
Chemistry, University of Lincolnshire, UK (e-mail: ttaylor@lincoln.ac.uk).
DOI: 10.7763/IJBBB.2014.V4.318
E. Moisture Analysis
Fish samples were dried using an oven for ±24 hours at
1000 50C until the weight of samples remained constant.
The percentage of moisture content was calculated based on
wet weight and multiplied by 100% [7].
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International Journal of Bioscience, Biochemistry and Bioinformatics, Vol. 4, No. 2, March 2014
The temperature in the smoke generator was reached
450ºC. At 488ºC the pyrolysis process is fairly complete [9].
The completion of pyrolysis is depending on the smoke
sources. PLS produced a higher percentage liquid smoke
than CLS, due to the biological characteristics. These liquid
smokes are dark brown in colour, and their odour has been
described as very aromatic and pleasant smoky with
significant floral notes. Study by [9] showed that 100 g of
thyme (Thymus vulgariis) L powder plants produced 173 ml
of liquid smoke. They alsofound that coconut shell pyrolysis
produces 52,8% liquid smoke, 31,7% charcoal and ash and
15,4% of volatile gasses such as: CO2, CO, CH4 and
hydrocarbons.20 major carbonyl derivatives from cellulose
and hemicellulose thermal degradation were detected in the
volatile fraction the liquid smokes (Table II).[10] found that
liquid smoke compounds were dominated by: toluene, 2prophenol, methoxybenzena, 1,2-dichlorobenzene; 1,1bicyclohexyl; 2,4-bis (1,1-dimethylethyl) phenol, 1,2benzenedicarboxylic acid diisooctyl ester, 4-nonylphenol
and is oxazole. The chemical compound of liquid are vary
based on their specific characteristics, although the major
component are usually groups of phenols, acids, aldehydes,
ketones, furanes, etc.
Major component found in PLS were phenol; 2furanmethanol; 1,2-benzenediol; phenol 2-methoxy; and 2cyclopenten-1-one,2-hydroxy-3-methyl. Meanwhile, CLS
were dominated by phenol,2-methoxy; phenol,2,6,dimethoxy;
pyrazole,1,4-dimethyl;
phenol,4-ethyl-2methoxy; and 1,2-benzenediol,3-methyl. [9] reported that
the main components thyme liquid smoke were 2furancarboxaldehyde;
2-furanmethanol;
3-methyl-1,2cyclopentanedione; 1-acetoxy-propan-2-one and 3-methyl2butanone. Which are different from those in PLS and
CLSother compounds in smaller concentrations contribute
with similar flavour notes, such as: cyclopentanone;2methyl-2-cyclopenten-1-one;3-methyl-2cyclopenten-1-one
and 3-hydroxy-2-methyl-4H-pyan-4-one.
F. Benzo (a) Pyrene Analysis
5g of sample was destructed by sodium sulphate and
chloroform until homogenious. The sample was then
extracted into propylene carbonat and saponificated with
NaOH solution. The extract was filtered with calcium
chloride, celite and florisil. The filtrate then evaporated to 1
ml using a water bath at 45oC, and passed a glass column
(clean up) containing 10% of aluminium oxide in the lower
part and 10 g of Na2S04 in the upper partuntil the component
was elured. Samples were then fractionated in the column in
silica 60 reins column, 70-230 mesh and added by 10%
diethylether and evaporated. Finally, the sample was
injected into Gas Chromatography to measure benzo ()
pyrenecontent [8].
G. Microbial Analysis
Fish samples were diluted into concentration of 10-2, 10-3,
10-4, and 10-5. Petri dishes containing of diluted samples
were then incubated with the opposite position at 35o C for
48 hours. The number of colonies was then calculated by
hand tally counter using for the plate amount of 25-250
colonies [7].
H. Sensory Characteristics
Sensory characteristics of the fish were evaluated using a
9-point hedonic scale for: general appearance, colour, odour,
flavour, and texture. Seven expert panelists were used to
evaluate the fish as described by Indonesian National
Standard.
I. Statistical Analysis
The collected data from two replications was subjected to
a t –test to determine the differences between PLS and CLS
(independent variable) for each parameter using the SPSS
statistical package. Significantly different treatment means
were further separated using the Least Significant
Differences method and significance was reported at P<0.05
and P<0.01 with means ±deviation standard.
B. Phenolic Compounds in Liquid Smoke
Total phenol derivatives formed from the thermal
degradation of lignin are also present and in general
contribute with typical smoky notes. These fraction are
important for their attributed antioxidant activity. The total
phenolic derivatives found in PLS were 25,1%, lower than
that of CLS of 26,31%. 36,0% [9] found that thyme liquid
smoke was total phenolic derivatives.
III. RESULTS AND DISCUSSION
A. Production of Liquid Smoke Flavouring
The data obtained on the production of liquid smoke are
presented in Table I. The materials for smoke sources in
Indonesia wereselected based on their availability in the
area. PLS and CLS can be found easily almost in all
provinces.
C. pH Value of Liquid Smoke
The pH value of PLS was 3,2; slightly higher than than
the CLS of 2,5. According to [10], the vapour phase of
smoke can be separated into acids and phenols which are
strongly acidic in water. Some commercial liquid smoke
have the pH value between 3-4 [11]. pH value  4 can
inhibit the growth of bacteria and moulds.
TABLE I: PRODUCTION OF LIQUID SMOKE
Liquid smoke material
Parameter
PLS
CLS
Material weight (kg)
2,5
2,5
1,375
1,15
Persentage of liquid smoke (%)
55
46
Charcoal & evaporated component (%)
45
54
Volume of liquid smoke (L)
D. Moisture Analysis of Smoked Skipjack
No statistically significant (P>0.05) differences were
observed in chemical properties in the samples produced by
both PLS and CLS. The moisture content of fresh skipjack
was 73.73% and immediately reduced caused by smoking
process.
The moisture content of both PLS (54.0±2.7) and CLS
(54.3±0.9) samples were in the range of standard Indonesian
smoked fish, which is 60% of maximum. Moisture content
Note:
PLS = Paddy chaff liquid smoke treatment
CLS = Coconut shell liquid smoke treatment
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International Journal of Bioscience, Biochemistry and Bioinformatics, Vol. 4, No. 2, March 2014
in this study had been decreased for about 26.3-26.9%
during smoking process. The reduction of moisture content
No
was also influenced by the presence of salt prior smoking
and its concentration.
TABLE II: PERCENTAGE OF MAJOR CARBONYL DERIVATIVES IN PADDY CHAFF AND COCONUT SHELL LIQUID SMOKE
Carbonyl derivatives
PLS
CLS
1
1,2-Benzenediol
6.45
6.76
2
1,2-Benzenediol,3-methoxi
1.22
4.81
3
2-Cyclopenten-1-one,2-hidroxy-3-methyl
5.14
3.93
4
2-Cyclopenten-1-one,3-ethyl-2-hydroxi
1.48
-
5
2-Furanmethanol
6.98
3.84
6
2-Furanmethanol,tetrahidro
1.83
-
7
Phenol,2-Methoxi-4-methyl-
1.68
3.45
8
Phenol
7.43
-
9
Phenol,2,6-dimethoxi
2.79
8.99
10
Phenol,2-methoxi
6.29
12.69
11
Phenol,3-methyl
-
3.96
12
Phenol,4-ethyl-2-methoxi
2.88
7.17
13
Phenol,2-methyl
1.97
2.71
14
Phenol,4-methyl
3.16
-
15
Phenol,2-Methoxi-3-methyl-
-
-
16
Phenol,4-ethyl
2.02
-
17
2-Propanone,1-4-hidroxi-3-methoxiphenyl
1.16
-
18
Pyrazole,1,4-dimethil
-
8.05
19
1,2-Benzenedicarboxylic acid, diethyl ester
-
1.15
20
1,2-Benzenediol,3-methyl-
1.4
4.78
Note: PLS = Paddy chaff liquid smoke treatment; CLS = Coconut shell liquid smoke treatment
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International Journal of Bioscience, Biochemistry and Bioinformatics, Vol. 4, No. 2, March 2014
TABLE III: SENSORY VALUE OF FRESH SKPIJACK
Spesification
Value±SD
E. Analysis of Benzo (α) Pyrene
Several authors have studied the carcinogenic activity of
liquid smoke. Although many smoke flavouring are free of
harmful compounds such as polycyclic aromatic
hydrocarbons (PAHs), but the presence of benzo () pyrene
(BP) in smoked fish has becoming a great consideration
nowadays. PAH constitute a large class of organic
substances containing two or more fused aromatic rings
made up of carbons and hydrogen atoms [12].
BP was the first PAHs to be identified as carcinogen, and
as a consequence, has been most studied. Benzo (α) pyrene
content in the PLS and CLS was not detected. While PLS
and CLS smoked skipjack was containing BP of 9.55 and
8.72 ppm respectively. Many researchers reported that the
range of BP found in smoked fish is about 1,7 - 53 ppm.
Therefore, efforts are needed to decrease the presence of BP
in smoked fish. This can be done by research as reputed
here of (using various free BP smoke sources) and also by
reducing the temperature of pyrolysis < 400°C.
Eye
8.00  0.67
Gill
8.00  0.67
Flesh and belly
8.20  0.63
Texture
8.50  0.53
Odour
8.20  0.63
Slime
9.00  0.00
Average
8.32  0.52
Note: SD = deviation standard
TABLE IV: EFFECTS OF LIQUID SMOKE ON SENSORY VALUE OF SKIPJACK,
ASSESSED ON FIRST DAY OF STORAGE
Sensory value±SD
Specification
PLS
CLS
F. Microbiological Analysis
Microbiological analysis showed that the total number of
colony of fresh fish was: 2x104cfu/g. The microorganisms
counted on all samples decrease with the application of the
smoking process. Very significant (P<0.01) differences
were detected between PLS and CLS samples. The average
of TVC of smoked skipjack being: 153cfu/g (PLS) and 76
cfu/g (CLS). In the Indonesian National Standard, the upper
limited for TPC either for fresh and smoked fish are:
5×105cfu/g. Therefore, from the microbiological point of
view the use of paddy chaff and coconut shell liquid smoke
are possible alternatives to traditional smoking method.
The results proved that liquid smoke reduced the
microbiological properties of the final product. According to
[13] liquid smoke has given an effect on the growth of the
Lb. plantarum. Study by [14] showed that at the beginning
(0 day) liquid and traditionally smoked beef tongues
samples had similar microbiological result namely 1.08-1.10,
1.55-1.70, 1.00-1.05, 1.09-1.05 and 2.14-2.20 log cfu/g for
total staphylococci/micrococci, APC (Total aerobic
bacteria), psychotrophs, halophiles and lactic acid bacteria,
respectively. According to [13] two components that
responsible for the bactericidal effect are phenolic and
organic acids the combination of them can effectively
control the growth of microorganisms.
Appearance
7.6±0.23
8.0±0.00
Odour
7.0±0.00
7.7±0.00
Flavour
7.7±0.00
7.7±0.00
Texure
7.0±0.00
7.0±0.00
Moulds
9.0±0.00
9.0±0.00
Average
7.7±0.05
7.8±0.00
Note:
PLS = Paddy chaff liquid smoke treatment
CLS = Coconut shell liquid smoke treatment
CLS smoked skipjack samples received higher scores in
appearance and odourcompared to PLS, while flavour,
texture and the present of moulds of both samples are
remain the same. The differences were not significant
(P>0.05). This is indicates that both treatment were
acceptable by the panelists in term of their sensory point of
view. Although the CLS samples gave slightly better result
than the PLS samples. Both samples were accepted by
panelists until 4th days of storage.
Smoking has been reported to affect the colour of food
[15], [16]. The formation of the smoke colour is believed to
originate from an uptake of coloured smoke constituents,
oxidation and polymerization of smoke compounds, and
reaction of smoke compounds with proteins. Therefore,
condensation reactions take place between carbonyls and
amines, leading to the appearance of the typical smoke
colour [17].
The CLS showed lower odour intensity and lower
brightness than the PLS. This indicates that there to be a
relationship between liquid smoke composition and the
different type of smoke sources used to produce it. [18]
indicated that colour development depended mainly on
creation between carbonyls, protein, and lipid, while the
flavour afforded was largely due to the type and amounts of
phenolic compounds present. These compounds also
influence the antioxidant effect of the liquid smoke.
The effect of liquid smoke on the textural attributes of
skipjack showed that CLS was firmless compared to PLS.
Although PLS was richer in carbonyls than that of CLS.
G. Sensory Value of Fresh Skipjack
An analysis of raw material of fish was carried out with
the effects of scores of samples given by 7 panellists. The
main results are presented in Table III.
Raw fish used in this study had the following
characteristics of eyes perfectly fresh, convex black pupil,
translucent cornea, bright red gills, no bacterial slime, outer
slime water white or transparent, bright opalescent sheen, no
bleacing. The value was in accordance with SNI standard
with the minimum sensory value of 7,0.
H. Sensory Value of Smoked Skipjack
In general, smoked skipjack treated by PLS received
slightly higher sensory scores than CLS treatment (see
Table IV).
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International Journal of Bioscience, Biochemistry and Bioinformatics, Vol. 4, No. 2, March 2014
[15] E. Marit and B. R. Anna Maria, ”Quality of Cold Smoked Salmon
This is largely caused by the reaction between carbonyls
and proteins [17].
Taking only sensory qualities into account, the samples
were rejected by the assessing panel when they showed
signs of softening, low elasticity, firmness, discolouration
and low brightness. All samples were rejected at 4 days of
storage. These results differed slightly bit from initial
research that with CLS and PLS. The shelf life of smoked
skipjack at room temperature was accepted by panelist until
6 days of storage.
offered to the General French consumer,” National Institute of
Nutrition and Seafood Reseach, 2004.
[16] O. Martinez, J. Salmeron, M. D.Guillén, and C. Casas, “Sensorial and
physicochemical characteristics of salmon (Salmosalar) treated by
different smoking process during storage,” Food Science and
Technology International, vol. 13, pp. 477, 2007.
[17] L. Toth and Potthast, “Chemical aspect of the smoking of meat
products,” Advanced in Food Research, vol. 27, pp. 87-158, London:
Academic Press.
[18] V. Vincent, C. Prost, and T. Serot, “Volatile aldehydes in smoked fish;
analysis methods, occurrence and mechanisms of formation,” Food
Chemistry, vol. 105, pp. 1536-1556, 2007.
Fronthea Swastawati was born on February 23th
1959 in Kebumen, Central Java, Indonesia. She
received the BS degree in Fisheries and Marine
Science from the University of Diponegoro,
Semarang, Indonesia in 1983, the MSc degree in Food
Science from the University Humberside, UK in 1993,
and the Doctoral degree in Coastal Resources
Management from the University of Diponegoro,
Semarang, Indonesia in 2008.
She is a lecturer of Fisheries and Marine Science at University of
Diponegoro, Semarang, Indonesia. She has published more than 20 papers
in journals, conference proceedings, and books. Her research interests
include Post Harvest Fisheries Technology.
Dr. Swastawati is a member of Asia-Pacific Chemical, Biological, and
Environmental Engineering Society. She received a The Most Excellent
Paper Award and Session Chair Participate on International Conference on
Biotechnology and Food Science in Bangkok, Thailand April 7-8, 2012.
IV. CONCLUSION
In conclusion, the two liquid smoke flavourings studied
led to desirable changes in the characteristics and safety of
skipjack flesh. These changes were similar in both PLS and
CLS treatments. Although each liquid smoke had specific
chemical compounds both could be used to smoke skipjack
and produce a satisfied smoked fish produced.
ACKNOWLEDGEMENT
This project is funded by Directorate General of Higher
Education, Indonesian Ministry of National Education.
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Y. S. Darmanto was born on October 17th 1951 in
Klaten, Central Java Indonesia. He recieves BS
degree in fisheries, Faculty of Animal, Diponegoro
University in 1978. He received master of science in
1985 in fisheries science and Ph.D degree in 1997 in
marine science and engineering from Nagasaki
University, Japan.
He is currently the university professor of Fisheries
by Product at Diponegoro University, Semarang Indonesia and HEAD of
Fisheries Processing Product Technology Laboratory Faculty of Fisheries
and Marine Science Diponegoro University.He applied this knowledge in
some research such as The effect of some fish bones collagen in improving
food quality; Effect of collagen from different types of fish bone to
miofibril protein quality and Physico chemical properties of some dried
fish product in Indonesia. He has published more than 30 papers in journals,
conference proceedings, and books.
Prof. Darmanto is a member of Indonesian Toxicology Society and
Indonesian Food Technologist Society.
Lachmuddin Sya’rani was born on July 18th 1939 in
Bengkulu. He received bachelor degree from National
University, Jakarta Indonesia in 1965. Receive Master
of Science and Ph.D degree in 1981 and 1983 from
Faculty of Science The University of Newcastle Upon
Tyne, UK. He has received professor in the field of
Marine Science.
Since he retirement from full time employment he
has continued to work at the doctoral degree in Coastal Resources
Management Diponegoro University, Semarang Indonesia. He has
supervised more than 25M.Sc/Ph.D projects to successful completion and
published more than 30 papers in refereed journals. He has done many
research,some of them Temporal changes on the reef flat of pulau pari
complex; The significance reef for archipelago country and solution of
them; Coral form and Acropora aspera (DANA) growth in the Java sea on
west season.
Prof. Sya’rani is a member of Asean Fisheries, Indonesian
Oceanographers Society, Indonesian Fisheries Scientist Society and many
other organizations both National and International.
Kapti Rahayu Kuswanto was born on November
15th 1944 in Tuban, East Java, Indonesia. She
received the BS degree in Food Technology from
Gadjah Mada University in 1971, receive Diploma
in Microbiology and Bioengineering, Osaka
University, Japan, 1975. She received Ph.D degree
in Tokyo University of Agriculture, Tokyo Japan,
1988.
She is currently the university professor of Food
International Journal of Bioscience, Biochemistry and Bioinformatics, Vol. 4, No. 2, March 2014
Technology and Microbiology at GadjahMadaUniversity, Yogyakarta
Indonesia.Her current research interests include food technology and
microbiology. She has published more than 30 technical papers in journals,
conference proceedings, and books.
Prof. Kuswanto received a JSPS for Completion of Ph.D Program at
Tokyo University of Agriculture Award.
degrees at eight UK Universities. Both his B.Sc and Ph.D were in
biochemistry, but when he moved to Grimsby in 1974, he applied this
knowledge of biochemistry to food, and in particular to post-harvest fish
research. Aspects of pure research included mechanism of blackspot
development and quality parameters, biochemical indices and nutritional
quality changes in fish during processing and storage. Applied aspects
include improved utilisation of waste and assessment and reduction of
losses in cured fish processing. He has worked extensively in South East
Asia (Indonesia, Malaysia, Thailand) as well as in Europe, USA, Kenya
and China.
Prof Anthony Taylor was awarded the Chair of Food Science in 1992,
one of the first in what was then Humberside Polytechnic. Since retirment
from full-time employment, he has continued to work on a part-time basis
at the Holbeach Campus.
K..D. Anthony Taylor was a senior academic and
head of Food Research Centre. He has supervised
more than 35 M.Phil/Ph.D projects to successful
completion and published more than 45 papers in
refereed journals.
He is in the Society of Chemical Industry for
which he organized a series of conferences on Fish
Processing, and was Chairman of SCI Food
Commodities and Ingredients Group. As well as being an External
examiner for B.Sc. and M.Sc.s in Food he has examined for research
99