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USING GC-MS TECHNIQUE TO EVALUATE THE OMEGA-3
CONTENT FROM OIL OF OF THREE SPECIES OF IRAQI
MARINE FISHES
Nada Rafid Othman*, Amjed KadhumResen**, Dhia Faleh Al fkaiki**
*Department of Fisheries and Marine Resources, College of Agriculture, University
of Basrah , Basrah, Iraq
** Department of Food Science, college of Agriculture College,University of Basrah,
Basrah , Iraq
(Received 26 August2015,Accepted 6 September 2016)
Keywords: Omega , Cynoglossusarels, Winter.
ABSTRACT
The current study was conducted from October 2013 until the beginning of
September 2014 to estimate the Omega -3fatty acid for three species of marine fish
(Acanthopagrusarabicus , Chelonsubviridis and Cynoglossusarels). The study was
conducted during the four seasons winter of 2013, spring of 2014 , summer of 2014
and fall of 2014. The oil was extracted by organic solvent extraction, the fatty acid
was estimated using Gas Chromatography connected with mass spectrometry
(calculated on the basis of the total amount of the fatty acids), omega–3 percentage of
both types DHA, EPA has varied between the fish as it reached 56.5% , 55.9% ,
67,13% , 49.22% during winter, spring, summer and autumn respectively. Also it was
found ,by the results, that the percentage of omega -3 type EPA was high in the winter
as the percentage was 30.99%, followed by the summer as it was 30.42% and the
lowest rate was in the autumn as it was 20.33%, while the percentage of the second
type of omega -3 DHA was close during the four seasons, as the summer achieved the
highest percentage which was 36.71% and the lowest percentage was in the winter as
it reached 25.06%.
INTRODUCTION
Fish oils are good sources of fatty acids that are not synthesized in the human body.
Fatty acids in fish oil have a very special character compared with other sources, they
consist not only of essential fatty acids, but are also an important source of omega-3
fatty acids, especially Eicosapentaenoic acid (EPA, 20:5 n-3) and Docosahexaenoic
acid(DHA, 22:6 n-3) and they are the most important fatty acids that are commonly
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found in marine fish and arise from phytoplankton and seaweed, which are part of the
food chain for fish (1). They have a positive effect on humane health because the
lever does not have the ability to manufacture them (2). (3), showed that fatty acid
content of omega-3 type in shark was 2500 mg / 100 g and for this it was
recommended to be as good and rich sources of omega -3. Omega-3 has an effect in
deceasing triglycerides in blood by 20% through balancing between triglycerides
storage and its independence (4) and also in decreasing heart clots (5).
The study aimed to extract fish oils and to identify fatty acids type omga-3 found in
these fish, and knowing the suitable conditions to form the primarily essential fatty
acids that give nutritional value for fish.
MATERIALS AND METHODS
Field study (work):
Three types of marine fishAcanthopagrusarabicus,Chelonsubviridis
,Cynoglossusarels. The Samples were collected quarterly from the Iraqi marine waters
northwest of the Arabian Gulf, samples were placed in containers filled of ice.
Laboratory work:
Samples were brought to the lab and cleaned andwashing bywater tap, sensory tests
were conducted, and these tests included: (smell and color of the gills and the
appearance of the eyes and cohesion of the skin peels and textures and exterior) at a
laboratory temperature, these specifications were recorded and stated, according to
(6). The smell and colour of the oils extracted were checked depending on the sense
of smell and look, then the muscles were extracted and separated from the rest of the
parts of each type of the studied fish species .
The guts, heads, skin and bones were removed thereby we have models of the samples
of each type of fish, and also meat of each type of fish was mixed separately.
Extraction of oils:
The Oil was extracted from samples of each species of the studied fish using the
method of extraction of organic solvent followed by (7) after mincing the samples to
ensure that they are small enough.100 g were taken of the mincing sample of the fish
and put in a beaker and then distilled water added and the mixture was mixed by
electric mixer, then chloroform and methanol were added to the previous mixture and
naturalized by electric mixer on the speed of the 2000 circles for one minute the
homogenized mixture was transferred to tubes and transported to the centrifuge , then
the mixture separated into three layers, the upper layer of water (methanol layer and
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water), semi-solid layer (concentrated with protein) and lower layer (dissolved oil
layer in chloroform), and the upper layer was taken out using the dropper and
neglected, while the lower layer with semi-solid layer were filtered on filtration paper,
and then evaporated and the oil was collected in dry glass bottles and saved by
freezing or cooling, The oil produced by this method is known crude oil.
Diagnosing the fatty acids by GasChromatographytechnique connected with
mass spectrometry (GC – MS)
A similar esterification was conducted to the fatty acids according to the method
described in (8) later the fatty acids were diagnosed in the extracted oil of the types of
fish using Gas Chromatography device connected with mass spectrometry type
(GC – MS QP210 Ultra, ShimadzuApan) equipped with capillary pole type DB-MS 5
(5% phenyl, 95% methyl polysiloxane) as an inactive (static) phase with dimensions
(30 meters in length and a diameter of 0.32 and the thickness of static phase
0.25micrometer) , also High-purity helium gas was used 99.9.
The separation process was conducted in accordance with the thermal program GC on
50o for a minute then raised to 150°C for two minutes at a rate of 5o per minute and
then raised 280° at a rate 5o per minute , the temperature was stabilized on 280oC for a
minute. The injection process was carried out using automatic injector type AOC -
20is, SHIMADZU.
RESULTS AND DISCUSSION
Omega -3 amount of fatty acids was determined, it was in high percentages in some
species, and of significant change; as a result of the different types of fish and the
difference of fat composition in the areas of the fish's body that varied relatively with
each other.The differences that happened in the fatty acid percentagesaredue to the
condition of food and heat.
The results showed the percentage and the amount of omega–3 in both types which
got to, during the winter, 56.05% . The highest percentage of EPA was in A.arabicus,
reaching 11.6% and the lowest value was in C.subviridi 8.74%, while the percentage
of the DHA was close in all types. C.arels achieved the highest percentage of 8.77%
and the lowest percentage in A.arabicus 8.04%. This indicates to increase the
proportion of the EPA during the winter compared with DHA as shown in Figure (1).
It is less than what has (9) in a study on fish oils in Ireland and found that the amount
of fatty acid EPA in fish oils 18.2% to the type of fatty acid DHA 12.2%.
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Fig.(1) the percentage of omega- 3 in A. arabicus , C. subviridi and C.arelsduring
winter
But in the spring the proportion of omega - 3 was about 55.29%, the highest
percentage of EPA was in A.arabicus, reaching 12.64% and the lowest value was in
C.arels 4.55%, while the percentage of the DHA was the highest in C.arels which was
12.94% and the lowest value inA.arabicus 5.45%, as in Figure (2). The difference
between the percentage of the two types of omega-3 acid due to the nature of the food
and the season. These results agreed with (10).
Fig (2) Omega-3 percentage of both types A.arabicus , C. subviridiand C.arels
during spring season
Figure (3) shows omega- 3 percentage in both types of marine fish during the summer
where it reached 67.14%, which the highest percentage of omega–3 acid during the
four seasons, asC.arelsachieved the highest percentage of both types EPA and DHA,
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reached to 11.14%, 14.65% respectively, and the lowest value for the EPA was in
C.subviridi 9.53%, and the lowest proportion of DHA was found in A.arabicus 10.4%
and the difference in omega-3 content due to the feeding techniques and the type of
food consumed by the dish (11).
Fig (3) Omega-3 percentage of both types in A. arabicus , C. subviridi and C.arels
during summer season.
showed for the autumn that there was about 49.12% of omega – 3 of both types EPA,
DHA in the studied fish. The C.arels achieved the highest percentage of both types
EPA and DHA, reached to 11.14%, 14.65% respectively , and the lowest value for
both types was found in A.arabicus2.14% , 7.15% respectively as shown in fig (4).
This difference in omega - 3 percentage depends on the nature of fish food
(12).
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Fig (4) Omega-3 percentage of both types A. arabicus , C. subviridi and C.arels
during summer season
When Comparing marine fish content of omega-3 between seasons of the year we
find that the fish in the summer, with the highest content of omega-3 compared to
other seasons, as shown in Figure (5). C.arels has contained the highest percentage of
omega -3 type DHA during all seasons and the highest percentage of EPA during the
summer and autumn.
Fig (5) Omega-3 percentage of both types in A. arabicus , C. subviridi and
C.arelsduring the four seasons
These differences due to the season of the year (Jangaard, et al., 1967) and the
structure of food (13) or the type of fish (11) and the depth of fishing water (14).
These results were agreed with (15) data in terms of that edible marine fish in the
Arabian Gulf contain high amounts of unsaturated fatty acids, particularly fatty acids
of the type omega-3, and with the results of (16 ) concerning marine fish in Norway.
(17) showed in their study about the quality of fatty acids in cooked and frozen
sardines that there are significant differences even between the same class of sardines.
These results agreed with (15) in his study of the importance of the nutrition of the
edible marine fish in Qatar Coast region, and those results were confirmed by (18),
who studied the fat content, fatty acids, cholesterol and the amount of energy in some
kind of fresh and cooked marine fish in the United Arab Emirate, and with (19) of
omega-3 content in some marine fish in Turkey, and also with (20) in determining the
type and structure of fatty acids in some marine fish in Turkey, also with (21) of the
content and structure of fatty acids in marine fish on the south west coast of Brazil.
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لتقییم محتوى الاومیکا- 3 فیزیوت ثلاث انواع من الاسماک البحریة GC- MS استخدام تقنیة
العراقیة
ندى رافد عثمان، أمجد کاظم رسن، ضیاء فالح الفکیکی*
قسم الاسماک والثروة البحریة ، کلیة الز ا رعة ، جامعة البصرة ، البصرة ، الع ا رق
*قسم علوم الاغذیة ، کلیة الز ا رعة ، جامعة البصرة ، البصرة ، الع ا رق
الخلاصھ
اجریت الدراسة الحالیة من بدایة تشرین الاول 2013 ولغایة ایلول 2014 لتقدیر الاحماض الدھنیة نوع اومیکا- 3
والبیاح الاخضر Acanthopagrusarabicus لثلاثة انواع من الاسماک البحریةوھی الشانک
اجریت الدراسة اثناء الفصول الاربعة ، Cynoglossusarels ولسان الثور الاملس Chelonsubviridis
شتاء 2013 و ربیع 2014 وصیف 2014 وخریف 2014 . تم استخلاص الزیت بطریقة الإستخلاص بالمذیب
العضوی ، وقدرت الأحماض الدھنیة بإستخدام کرماتوکرافی الغاز المتصل بمطیاف الکتلة (محسوبة على أساس
EPA , المجوع الکلی للأحماض الدھنیة الکلیة) ، وقد تباینت نسبة الاومیکا- 3 بین الانواع السمکیة بنوعیھا
49.22 % خلال فصل الشتاء ، الربیع ، الصیف ، %67.13 ، % 55.29 ، % اذ بلغت 56.05 DHA
کانت مرتفعة فی فصل EPA والخریف على التوالی. کذلک وجد من خلال النتائج ان نسبة الاومیکا- 3 نوع
الشتاء اذ بلغت نسبتھا 30.99 % ویلیھا فصل الصیف اذ کانت 30.42 % واقل نسبة کانت فی فصل الخریف اذ
فکانت نسبتھا متقاربھ خلال الفصول الاربعة اذ حقق فصل DHA بلغت 20.33 % ، اما النوع الثانی للاومیکا- 3
. % الصیف اعلى نسبة لھا وھی 36.71 % وادنى نسبة کانت فی فصل الشتاء اذ بلغت 25.06
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