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Dietary Soybean Phospholipid Increase The Oxidate Stability of Lipids Extracted From Fish Fillets
Dietary Soybean Phospholipid Increase The Oxidate Stability of Lipids Extracted From Fish Fillets
The Nisshin OilliO Group, Ltd. (1 Shinmeicho, Yokosuka, Kanagawa 239-0832, JAPAN)
Department of Food and Nutrition, Toyama College (444 Gankaiji, Toyama 093-0193, JAPAN)
3
Faculty of Agriculture, Kagawa University (2393 Ikenobe, Miki-cho, Kita-gun, Kagawa 761-0795, JAPAN)
2
Abstract: We previously reported that the feeding of soybean phospholipids to fish increased the storage
stability of n-3 polyunsaturated fatty acids (PUFA)-rich fish fillets. In this study, we examined the storage
stability of lipids extracted from fish fed a diet containing soybean phospholipids and fish oil. Rainbow trout
were divided into two groups, and were fed an either 2.5% soybean phospholipids (test) or no phospholipids
(control) containing diet for 4 weeks. Lipids were extracted from fish fillets after the feeding period, and
were subjected to an oxidation test. Lipids extracted from the fillets of fish in the test group exhibited lower
values of oxygen absorption than those in the control group, and the degradation of docosahexaenoic acid
(DHA) and eicosapentaenoic acid (EPA) was inhibited. Higher percentages of DHA and EPA were bound to
phosphatidylcholine and phosphatidylethanolamine in the extracted lipids in the test group than in the
control group. These results indicate that the oxidative stability of lipids extracted from fish fed soybean
phospholipids is high, and that the higher percentages of DHA and EPA in PC and PE may have resulted in
the higher stability of the lipids extracted from fish fillet.
Key words: soybean phospholipids, fish fillet, lipid, oxidation, phosphatidylcholine, phosphatidylethanolamine,
docosahexaenoic acid, eicosapentaenoic acid
1 INTRODUCTION
We previously reported that the feeding of soybean
phospholipids to rainbow trout increased the storage stability of fish fillets1. Soybean phospholipid feeding did not
influence the level or fatty acid composition of total lipids,
or the content of antioxidants, tocopherol and phospholipid in fish fillets. Rainbow trout fed a soybean phospholipid
diet had a higher percentage of docosahexaenoic acid
DHAin phosphatidylcholinePCand phosphatidylethanolaminePEin fillets than those fed a soybean phospholipid-free diet2. The oxidative stability of DHA-bound PC
and PE has been reported to be high3, 4. We speculate that
one of the factors associated with the increased storage
stability of fish fillets was the high percentage of DHA were
bound to PC and PE. However, in addition to lipids, watersoluble vitamins, amino acids, and proteins may also influence the storage stability of fish fillets 5, 6. If the high
amounts of DHA-bound PC and PE increased the storage
stability of fish fillets, the oxidative stability of lipids ex-
Correspondence to: Yoshihiro Murano, The Nisshin OilliO Group, Ltd., 1 Shinmeicho, Yokosuka, Kanagawa 239-0832, JAPAN
E-mail : y-murano@nisshin-oillio.com
Accepted March 4, 2010 (received for review January 28, 2010)
2 EXPERIMENTAL
2.1 Materials
The soybean phospholipids and soybean oil used in this
study were provided by Nisshin OilliO Group Ltd., Tokyo,
Japan. The fish oil was obtained Nippon Suisan Kaisha,
Ltd., Tokyo, Japan.
2.2 Procedure
2.2.1 Fish and feeding experiment
A purified diet, the composition of which is shown in
Table 1, was used in the experiment. Vitamins and minerals were added, as described by Kaushik et al.9and Ogino
et al.10. Two types of experimental diet containingtest
dietand not containing soybean phospholipidscontrol
dietwere prepared as follows. Fish oil10 g/100 gwas
added to each diet. In addition, soybean oil
5 g/100 gwas
added to the control diet while soybean phospholipids
2.5
g/100 gand 2.5 g of soybean oil were added to the test
diet. The fatty acid composition of the total lipids in each
type of experimental diet is shown in Table 2.
Test fish were maintained and fed as previously described 1. Juvenile rainbow trout Onchorhynchus
mykisspurchased from a local supplier were used in the
feeding experiment. At the start of the feeding experiment,
a total of 140 fisheach weighing 2.1 g on average
were divided into 2 groups70 fish per group. At the end of the
feeding period, fish fillets were collected as previously described, and used as samples.
2.2.2 Biochemical analysis
Lipids were extracted from samples according to the
method of Folch et al.11. The phospholipid content, fatty
acid composition, and tocopherol content were determined
as previously described1. Triacylglycerols were fractionated by thin-layer chromatography on Silicagel 70 plates
Wako Pure Chemical Industries, Ltd., Osaka, Japanusing
Test
(g/100 g)
Casein
50
Dextrin
20
Corn starch
Mineral mix
Vitamin mix
Lipids
Fish oil
10
10
Soybean oil
2.5
Soybean phospholipids
2.5
Test
(g/100 g)
12:0 *
0.1
0.1
14:0
4.0
4.0
15:0
0.3
0.4
16:0
12.7
14.2
16:1
4.9
4.9
17:0
0.2
0.3
18:0
3.1
3.1
18:1
21.4
19.0
18:2 (n-6)
18.3
19.2
18:3 (n-3)
2.7
2.8
20:0
0.3
0.2
20:1
9.0
8.9
20:2 (n-6)
0.2
0.2
22:0
0.6
0.6
22:1
8.0
8.0
20:5 (n-3)
7.1
7.1
24:0
0.4
0.4
24:1
0.5
0.5
22:5 (n-3)
1.0
1.0
22:6 (n-3)
5.2
5.2
458
22:6
were measured as indicators of the storage stability of
extracted lipids. The oxygen concentration in the headspace of each vial was analyzed with an oxygen meter
RO-102; Iijima Electronics, Corp. Aichi, Japan
. Fatty acids in each vial were quantified by gas chromatography.
2.2.4 Statistical analysis
Data are expressed as the meanSEM. Significant differences between the diet groups were determined by Students t-test. Differences with a P-value 0.05 were considered significant. A statistical software packageToukei
version 2, Esumi, Tokyo, Japanwas used for all statistical
analyses.
Test
(g/100 g)
3 RESULTS
Table 3 shows the growth of rainbow trout fed a control
or test diet for 4 weeks. There was no significant difference
in body weight or feed conversion efficiency between the
test and control groups. The supplementation of soybean
phospholipids did not influence growth.
The total lipid content of fish fillets after the feeding period in the control and test groups was 5.90.0 and 6.0
0.1g/100 g, respectively, showing no significant difference. There was no significant difference in the content of
triacylglycerol, phospholipids, PC, PE or tocopherols in the
Test
2.1 0.0
2.1 0.0
6.2 0.2
6.2 0.2
118.0
118.5
14:0*
4.1 0.1
4.0 0.1
16:0
19.7 0.3
20.5 0.2
16:1
7.7 0.2
7.6 0.1
18:0
4.2 0.1
4.2 0.0
18:1
27.1 0.4
26.8 0.2
18:2 (n-6)
11.9 0.7
10.4 0.5
18:3 (n-3)
1.4 0.0
1.3 0.0
20:1
6.4 0.3
6.9 0.4
20:2 (n-6)
0.6 0.0
0.6 0.1
22:0
0.5 0.0
0.6 0.0
22:1
3.9 0.3
4.3 0.2
20:5 (n-3)
2.7 0.1
2.8 0.1
22:5 (n-3)
0.8 0.0
0.8 0.0
22:6 (n-3)
7.5 0.2
7.9 0.0
Test
69.6 2.1
68.2 4.7
16.9 0.5
16.7 0.9
12.5 0.3
12.7 0.5
Phosphatidylethanolamine
(g/100 g lipid)
4.2 0.2
4.0 0.5
0.43 0.04
0.43 0.03
Fig. 2 Effects of Dietary Soybean Phospholipids on the Stability of Extracted Lipids from Fish Fillets during the
Oxidation Period.
The oxidative stability was evaluated by the residual fatty acid content based on lipid oxidation. Values are means
SEM (n=3). Asterisks indicate significant differences (P<0.05).
dation. No significant difference was noted in the fatty acid
composition of extracted lipids used in the oxidation test
Table 5. Measurement of the oxygen concentration in
the headspace during the oxidation test showed that the
amount of oxygen absorption on day 10 or later was significantly lowers in the test than in the control group
Fig. 1
.
Analysis of the amount of fatty acids remaining during the
oxidation test revealed no significant difference in the
amount of palmitic or oleic acid between the test and control groups. In contrast, the amounts of residual DHA and
EPA decreased with the duration of storage in both groups,
but were significantly higher in the test than in the control
group, indicating the inhibition of fatty acid oxidation in
the test group
Fig. 2.
Table 6 shows the fatty acid composition of triacylglycerol in lipids from fish fillets. The amount of 22:1 in triacylglycerol was significantly higher in the test group than the
control group, however, the amounts of DHA and EPA
were not significantly different between the two groups.
Analysis of the amount of fatty acids in PC and PE showed
that DHA and EPA were bound predominantly to the sn-2
position of PC in both groups; larger amounts of DHA and
EPA were bound to the sn-1 and sn-2 positions of PC in the
test than in the control group
Fig. 3 A, B
; a significantly
higher percentage of DHA and EPA were bound to the sn-1
position of PE in the test than in the control group
Fig. 4
A; and significantly higher percentages of DHA was bound
Test
(g/100g)
14:0*
4.0 0.0
4.1 0.1
16:0
20.3 0.2
20.4 0.5
16:1
7.9 0.3
7.9 0.2
18:0
4.6 0.1
4.6 0.2
18:1
29.9 0.5
28.9 0.6
18:2(n-6)
11.5 0.2
10.5 0.4
18:3(n-3)
1.1 0.1
1.2 0.1
20:1
6.8 0.1
7.2 0.1
20:2(n-6)
0.6 0.0
0.6 0.0
22:1
4.9 0.1a
5.3 0.1b
20:5(n-3)
1.5 0.1
1.8 0.1
24:1
0.5 0.0
0.5 0.0
22:5(n-3)
0.6 0.0
0.7 0.0
22:6(n-3)
3.7 0.3
4.3 0.3
460
Fig. 4 Fatty Acid Composition of Phosphatidylethanolamine at sn-1 (A) and sn-2 (B) Positions of Lipids
from Fillets.
Values are means SEM (n=5). Asterisks indicate
significant differences (P<0.05).
to the sn-2 position of PE in the test than in the control
group
Fig. 4 B.
4 DISCUSSION
The oxidation test on lipids extracted from fish fillets
showed that the amount of oxygen absorption was significantly lower, and the amounts of residual DHA and EPA
were significantly higher, in the test than in the control
group. These results indicate that lipids contained in the
fillets of soybean phospholipid-fed fish exhibit a higher oxidative stability than those of soybean oil-fed fish. The analysis of lipids extracted from fish fillets revealed no significant difference in the content of phospholipids or
tocopherols, but higher percentages of DHA and EPA were
bound to PC and PE in extracted lipids in the test than in
the control group.
We previously reported that the feeding of soybean
phospholipids to rainbow trout increased the storage stability of fish fillets1, 2. The present study showed that the
feeding of fish with soybean phospholipids increased the
oxidative stability of lipids extracted from them, thus con-
461
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