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Seasonal Changes in Intrinsic Characteristics of Pacific Sardine (Sardinopssagax)Tomoko Okadaa; Michael T. Morrisseyaa OSU Seafood Laboratory, Astoria, OR

To cite this Article Okada, Tomoko and Morrissey, Michael T.(2007) 'Seasonal Changes in Intrinsic Characteristics ofPacific Sardine (Sardinops sagax)', Journal of Aquatic Food Product Technology, 16: 1, 51 71

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Seasonal Changesin Intrinsic Characteristics

of Pacific Sardine (Sardinops sagax)

Tomoko OkadaMichael T. Morrissey

ABSTRACT. Intrinsic characteristics of sardines harvested off theOregon-Washington coast were determined. The lipid content was ini-tially low (6.79%) in the beginning of the season in June, significantly in-creased to 22.95% in mid-August, and decreased slightly by the end ofSeptember. An inverse correlation (R2 = 0.90) was found betweenlipidand moisture content. Triacylglycerides werethe predominant lipidfraction, while the phospholipids fraction was the lowest. Analysisshowed that 20:5n3 was the most abundant fatty acid in sardine oil fol-lowed by 16:0 and 22:6n3 fatty acids. The level of 20:5n3 fatty acidwas slightly higher in August-September than earlier in the season.doi:10.1300/J030v16n01_05 [Article copies available for a fee from The Haworth

Document Delivery Service: 1-800-HAWORTH. E-mail address: Website: 2007 by The

Haworth Press, Inc. All rights reserved.]

KEYWORDS. Sardine, lipid, proximateanalysis, gas chromatography,thin layer chromatography

Tomoko Okada and Michael T. Morrissey are affiliated with OSU Seafood Labora-tory, 2001 Marine Drive, Room 253, Astoria, OR 97103.

This work was supported by grant No. NA16RG1039 from the National Oceanicand Atmospheric Administration to the Oregon State University Sea Grant CollegeProgram and by appropriations made by the Oregon State Legislature.

Journal of Aquatic Food Product Technology, Vol. 16(1) 2007Available online at http://jafpt.haworthpress.com

2007 by The Haworth Press, Inc. All rights reserved.doi:10.1300/J030v16n01_05 51
http://www.haworthpress.com/http://jafpt.haworthpress.com/http://jafpt.haworthpress.com/http://www.haworthpress.com/


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INTRODUCTION

Sardines are subtropical species existing in areas of subarctic andtropical ocean mixing, where they constitute some of the most volumi-nous fisheries species in the world (Leonardis and Macciola, 2004;Lluch- Belda et al., 1991). They are gregarious plankton-eating fish thatlive in large schools in the open sea and are usually found in the uppertrophic levels of the ocean (Leonardis and Macciola, 2004). The Pacificsardine (Sardinops sagax) is a coastal pelagic fish found from the Gulf ofCalifornia to southeastern Alaska (Hammann, 1998; Hammann et al.,1988; Schweigert, 2002). Pacific sardines were first landed commer-cially in thePacific Northwest during the 1935-36 season and continued

to be a very active fishery until collapsing in the early 1940s (Emmetet al., 2005). The abrupt decline in abundance of Pacific sardines islargely related to ocean temperature changes linked to climate or oceanregime changes that vary over periods of about 50 years (Chavez et al.,2003).

A new Pacific sardines fishery began in the Oregon-Washington(OR-WA) coastal area in 1999 and the catch volumeincreased from 776metric tons (mt) in 1999 to over 45,000 mt in 2005 [(Figure 1), (McCraeand Smith, 2006)]. Sardine landings generally begin in June and peak in

52 JOURNAL OF AQUATIC FOOD PRODUCT TECHNOLOGY

50

Landed sardines

Vessels targetingsardines

30

25

20

15

10

5

0

40

30

20

10

0 1999 2000 2001 2002

Year

Vessels

Landingsin1000metrictons

2003 2004 2005

FIGURE 1. Changes in sardine landings and vessels targeting sardines from1999 to 2005 in Oregon-Washington coastal area.


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August-September depending on catch rates and fishery quotas (Emmetet al., 2005).

Sardines and other pelagic fish are often termed fatty fish with lipidcontent that can reach levels in excess of 20% but can also drop below4%,depending on time of year (Caponio et al., 2004). This variability de-pends on several factors including seasonality, metabolic function, foodavailability and type, sea water temperature, maturity, spawning, andgeographic location (Aidos et al., 2002; Leonardis and Macciola, 2004).Sardines contain high levels of lipids and high amounts of n-3 long-chainpolyunsaturated fatty acids (n-3 LC-PUFA) including 20:5n3 and22:6n3, also known as eicosapentaenoic acid (EPA) and docosa-hexaenoic acid (DHA), respectively. These fatty acids play important

physiological roles in fish as components of membrane phospholipidsand as precursors of biologically active eicosanoids (Rodriguez et al.,2004).

The nutritional importance of fish oil for human nutrition is wellknown and is mainly associated with EPA and DHA. These fatty acidshave been shown to exhibit several beneficial effects on human healthincluding reduction of coronary heart disease, partial remediation of di-abetes, delay of the onset of Alzheimers disease, and inhibition of vari-ous types of cancers (Alasalvar et al., 2002; Holub and Holub, 2004;Nettleton and Katz, 2005). Recent work also has demonstrated positiveeffects of these fatty acids in the development of brain and nervoustissue for infants (Innis, 2003, 2004). It has been reported that n-3LC-PUFAs are effective in reducing the risk of behavioral and learning

disorders including attention deficit hyperactivity disorder, dyslexia,dyspraxia and autism (Ruxton et al., 2004)as well as mental illness suchas depression, suicidal tendencies and other violent behaviors (Hibbelnand Salem, 2001). Studies suggest that intake of n-3 LC-PUFAs hasbeneficial effects on symptoms of Alzheimers disease such as short-term memory loss, depressed mood, and inability to sleep, as well asend-stage Huntingtons disease (Haag, 2003; Tiemeier et al., 2003).

Although the sardine biomass for the U.S. fishery stretches along theWest Coast from Mexico to Alaska, the fishing activity is concentratedat the mouth of the Columbia River, which borders WA and OR. Sar-dine schools are located by fixed-wing airplane and harvested by largepurse seining vessels (25-40 m) which can capture 20-40 mt each set.The fish are pumped directly from the nets into refrigerated seawater

holds on board the vessels and rapidly cooled to < 4C (Dion, 2003).The fish are normally harvested in the afternoon and off-loaded at pro-cessing plants within 8 h of capture, graded by size, packed into 10 kg

Tomoko Okada and Michael T. Morrissey 53


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boxes and frozen. This allows for rapid turnover from harvests to finalproduct and helps maintain a very high quality. However, most landingsat Astoria, OR (located near the mouth of the Columbia River) have beenprocessed as bait for the overseas long-line tuna fishery with lowex-vessel prices of approximately $100-120/ton. Sardines landed inAstoria tend to be relatively large in size, with average weights of about150-180 g. While this has caused some concern in the bait-fish industry,which prefers smaller sardines (130-150 g), it does provide opportuni-ties for developing new products and directing these sardines towardhuman consumption.

The objective of this study was to determine intrinsic characteristicsin sardines harvested off the OR-WA coast during the 2004 season and

track changes in these characteristics as the season progressed. A betterunderstanding of seasonal changes that occur in theproximate composi-tion and other parameters of sardines will provide useful informationfor the growing sardine industry and assist its expansion into newmarkets.

MATERIALS AND METHODS

Fish and Sample Preparation

Sardines (Sardinops sagax) were harvested off the OR-WA coast bycommercial purse seiners and samples were collected and analyzed be-tween June 8 and September 23, 2004. The sardines were pumped fromthe seine nets directly into refrigerated sea water holds aboard the ves-sels and rapidly brought down to 0-4C. The sardines were off-loadedwithin 8 h of capture at local processing plants in Astoria, and separatedinto three different size groups. Seven of the smallest-, middle-, andlargest-sized sardines were randomly chosen, packed in ice and deliv-ered to the laboratoryon the day of catch. Analysis of the sardines beganthe following day and physical measurements (weight and length) weretaken. The head, viscera, and backbone were removed from each fish,and skin-on fillets were obtained. The fillets from all three sizes weremixed together and minced by a vertical-cutter/mixer (Stephan Food

Processing Technology, West Germany) for 2 min and kept in vacuum-sealed bags at 30C for further analysis. In a separate set of experi-ments, sardines were separated by weight (150-199.9 g and 200-249.9 g)



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during the period of July to August to determine if sardine size had aneffect on total lipid content.

Proximate Analysis

Moisture analysis was carried out according to Association of Offi-cial Analytical Chemists (AOAC) method 934.01 (1998). Total nitro-gen content was determined by Kjeldahl method according to AOACmethod 940.25 (1998) and multiplied by 6.25 to estimate the crude pro-tein content. Ash content was gravimetrically determined using a muf-fle furnace by heating at 515C to constant weight according to AOACmethod 938.08 (1998). Total lipid content was determined by hydrolyz-

ing the sample with hydrochloric acid in 100C water bath and extract-ing the lipid fraction with diethyl ether and petroleum ether. The lipidwas obtained after heating on 100C hot plate and in an oven to removethe ether layers according to AOAC method 948.15 (1998).

Thin-Layer Chromatography Separation

Extracted sardine lipids were subjected to thin layer chromatogra-phy (TLC) separation for lipid composition determination. Lipidswere separated into triacylglycerde (TG), diacylglyceride (DG), mono-acylglyceride (MG), free fatty acids (FFA), and phospholipids (PL).Samples were dissolved in hexane (10% solution, v/v), and an aliquotof

5 l sample mixture was loaded onto the silica plate (K6 silica gel 60,200 200 0.25 mm, Whatman Inc., Brentford, UK). The developingsolvent was hexane:diethylether:acetic acid (60:40:2, volume basis).Spots were visualized by spraying 5% sulfuric acid solution followedby drying at 100C on the hot plate for 30 min. Quantification was doneby densitometory ChemDoc with Quantity One software (Bio-Rad, Her-cules, CA). Commercially available TLC standard (Cat. No. 18-1A and18-5A, Nu Check Prep, Inc., Elysian, MN) was used for qualification,and fraction levels were expressed as w/w%.

Determination of Fatty Acid Compositionby Gas Chromatography

Fatty acids in sardine oil were converted into fatty acid methyl es-ters (FAME) according to AOAC method 991.39 (1998), and their



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composition was determined by gas chromatography (GC). A Hewlett-Packard 5890 Series II gas chromatograph (Palo Alto, CA) equippedwith a flame-ionization detector and capillary column (EC-wax, 30 m 0.25 mm i.d.; split ratio, 100:1; Alltech, Deerfield, IL) was used for ana-lyzing FAME. Parameters of the GC system were set as follows: injectorand detector temperatures 250C and 270C, respectively; the columntemperature was set to 50C and gradually heated to 180C at a rate of5C/min, then slowed to a rate of 0.8C/min until it reached 220C; he-lium was used as a carrier gas. The fatty acid concentrations were calcu-lated by comparison of their retention times with those of the referencestandards (Supelco, Bellefonte, PA).

Statistical Analysis

The results were presented as average and standard deviations ofeach experiment done in triplicates. Statistical comparisons were madeamong samples using ANOVA and Tukeys test; linear regression anal-yses were applied using S-PLUS software package. The results werepresented in terms of p values, with significant mean differences at thep < 0.05 level.

RESULTS AND DICUSSION

Physical Measurement

The weight and length of sardines collected for this study in Astoriaduring 2004 are shown in Figures 2 and 3, respectively. The average(ave) weight was 189.18 33.26 g, and the average length was 27.41 1.76 cm (n = 230) throughout the fishing season. This is within therange reported by Oregon Dept. of Fisheries and Wildlife for the sameseason [(31.3 g-293.6 g, ave 154.4 g), (McCrae and Smith, 2005)]. Fishsize depends on the year-class of the fish being harvested and the largesardines are associated with five-year old fish. Sardines from the Astoriaarea were larger in size compared to reports in regions such as Japan(Sardinops melanostictus), with a weight range of 30.6-90.7 g and

length range of 12.2-19.7 cm (Shirai et al., 2002); Turkey (Sardinapilchardus), with a range of 60-80 g in weight and 14-16 cm in length(Serdaroglu and Felekogglu, 2005); and Brazil (Sarinella sp.), with



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weight range of 48.6-98.4 g and length of 14.6-19.1 cm (Luzia et al.,2003). Pacific sardines from the Gulf of California, a similar species asthose in the present study, were also smaller [(18.3 0.4 cm; 112.0 12.6 g), (Ramirez-Suarez et al., 2000)]. The sardines in this study

showed consistent weight and length measurements early in the seasonand gradually showed a broader weight range. Emmett (2005) also re-ported that large- (> 18 cm) and medium-sized (14-16 cm) sardines


250

200

150

100

50

Weight(g)

06/12 6/22 7/2 7/12 7/22 8/1

Date

8/11 8/21 8/31 9/10 9/20 9/30

FIGURE 2. Weights of sardines collected in the Oregon-Washington coastalarea from June to September 2004.

35

30

25

20

15

10

5

06/12 6/22 7/2 7/12 7/22 8/1 8/11

Date

Length(cm)

8/21 8/31 9/10 9/20 9/30

FIGURE 3. Lengths of sardines collected in the Oregon-Washington coastalarea from June to September 2004.


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were found in June offshorenear Astoria, while small sardines (


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TABLE 1. Changes in proximate composition of sardines over summer 200

6/24 7/7 7/23 8/3 8/18 9

Moisture 68.58 0.25a 66.21 0.07b 61.53 0.19d 60.01 0.19f 60.52 0.22ef 60.94

Protein 17.70 0.26abc 17.81 0.13ab 18.33 0.48a 17.04 0.26cde 16.64 0.07de 16.32

Lipid 13.73 0.21a 13.30 0.34a 18.60 0.34b 19.87 0.18d 21.02 0.38e 19.38

Ash 1.68 0.03ab 1.60 0.11ab 1.63 0.06ab 1.76 0.11b 1.67 0.01ab 1.50

Values are % mean S.D. of twenty-one homogeneous samples consisting of seven sardines from each size category (100-150 g

The values in the same row labeled with a different superscript letter are significantly different (p 0.05).

59


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the spring and fall. The area between Point Conception and Ensenadais the major spawning area (Hernandez-Vazquez, 1994); however,spawning has also occurred on the southern Oregon coast in recentyears with the expansion of the biomass geographical area (Schweigert,2002). The low lipid content of sardines at the beginning of the season(June) is most likely due to the spawning season in the previous monthsresulting in fat mobilization associated with gametogenesis (Hardy andKeay, 1972). Studies have also shown that lower content of lipid in fishcan be observed in the reproductive season of the species since reducedfeeding level and sexual maturation contribute to reduced lipid levelsduring winter (Gokce et al., 2004). Lower lipid content of sardines at theend of season could be explained by a sexual maturation period towardsthe winter season and lipid migration from the flesh to the gonads.

Changes in moisture content were proportional to lipid content, andthe highest moisture level was found in June (72.50%), while the lowest(59.80%) was in August (Figure 4). Protein analysis showed the sardines


80

70

60

50

40

30

20

10

06/2 6/12 6/22 7/2 7/12

Date

Lipidandmoisture(%)

7/22 8/1 8/11

Lipid %

Moisture %

8/21 8/31 9/10 9/20 9/30

FIGURE 4. Lipid and moisture content of sardine over the 2004 fisheryseason.


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contained 16.32% to 18.33% crude protein, indicating sardines are agood source of protein. Ash content showed only a slight variation from1.50% to 1.76%. Results of proximate analysis were generally similar toother studies using Sardina pilchardus (Castrillon et al., 1997; Gokodluet al., 1998; Nunes et al., 1992; Serdaroglu and Felekogglu, 2005). Pro-tein and ash content remained constant throughout the season, account-ing for approximately 18-20% of the total composition (Table 1). Theprotein levels tend to be constant since they are mobilized from the fleshlate in the depletion process and the first to be restored after the comple-tion of spawning due to their structural importance (Love, 1997).

A reversecorrelation (R2 =0.90) was found between lipid and mois-ture content (Figure 5), which is in agreement with other sardine stud-ies such as Sardina pilchardus (Nunes et al., 1992) and Sardinopsmelanosticta (Sato et al., 1978). This correlation has also been found inother fish species including albacore tuna (Thunnus alalunga) (Wheelerand Morissey, 2003) and hoki (MacDonald et al., 2002). It has been sug-gested that lipid content is inversely proportional to moisture content inmost pelagic species (Castrillon et al., 1997; Love, 1997). A simple linear


25

20

15

10

5

055 60

Lipidcontent(%)

65

Moisture content (%)

70 75

FIGURE 5. Correlation between lipid and moisture of Pacific sardine.


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regression equation was developed that shows the relation between lipidand moisture as:

{lipidmoisture} = 35.23 1 (0.32) moisture %.

Changes in Lipid Composition

Changes in the lipid composition of sardines are shown in Table 2.Neutral lipids ranged from 97.43% to 98.56% with TG being the pre-dominant fraction varying from 78.62% to 80.93%. The lipid composi-tion levels remained constant throughout the season except free fatty

acids which varied from 5.05% to 7.15% with the highest occurringtowards the end of July (p < 0.05). DG and MG content ranged from4.30% to 5.54% and 2.02% to 2.76%, with relatively constant lev-els during the season. PL fractions existed in very low levels varyingslightly from 1.83% to 2.13%. It has been reported that fatty fishhad higher amounts of TG lipids compared with lean fish and loweramounts of free fatty acids, cholesterol, and polar lipids (Bandarraet al.,1997; Leonardis and Macciola, 2004). Leonardis and Macciola also re-ported different lipid composition in lean and fat sardine fillets anddemonstrated that PL were quantitatively equal in both lean and fattysardines (0.7 and 0.6 g/100 g sardines, respectively) where TG levelswere significantly higher (p < 0.05) in fatty sardines (12.1 g/100 g sar-dines) compared with lean sardines (1.3 g/100 g sardine). They sug-

gested that during hotter summer months, sardines accumulate reservelipids, especially TG, which is metabolized for energy during the coldseason. Fatty fish in the Bandarra et al. study (1997) contained 96.35%neutral lipids, which agrees with our study except that their studyshowed higher TG levels (91.63%). The difference could be due to en-vironmental and physiological differences including diet availabilityand the sexual maturity of sardines. Also, lipid content of lean fish intheir study was 1.20% which was considerably lower than the early sea-son fish in this study.

Changes in Fatty Acid Composition

The fatty acid composition of sardine lipids between June andSeptember is shown in Table 3. The major fatty acids in sardine lipidswere 16:0, 16:1n7, 20:5n3, and 22:6n3, with 20:5n3 being the highest



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TABLE 2. Changes in lipid composition of Pacific sardines.

6/24 7/7 7/23 8/3 8/18 9

Neutral lipids 98.05 0.29a 97.87 0.37a 98.14 0.40a 98.56 0.46a 97.43 0.78a 98.05

TG 78.62 0.52a 78.62 0.24a 78.81 0.28a 78.25 2.21a 79.26 1.76a 79.80

DG 4.84 0.39a 4.56 0.52a 4.87 0.61a 5.54 0.51a 4.52 0.60a 4.70

MG 2.02 0.91

a

2.19 0.19

a

2.41 0.68

a

2.76 0.90

a

2.66 0.58

a

2.63FFA 6.81 0.32ab 6.68 0.40abc 7.15 0.68a 6.41 0.56abc 5.83 0.99abc 5.34

Phospholipids 1.95 0.29a 2.13 0.37a 1.83 0.40a 1.83 0.52a 1.90 0.39a 1.95

Values are % mean S.D. of twenty-one homogeneous samples consisting of seven sardines from each size category (100-150 g

Values in the same row labeled with a different superscript letter are significantly different (p 0.05).

63


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TABLE 3. Changes in fatty acid composition (w/w %) of total sardine lipid.

FFA 6/24 7/7 7/23 8/3 8/18 9

C14:0 7.42 0.54a 7.09 0.04a 7.19 0.01a 7.33 0.09a 7.59 0.34a 7.57

C15:0 0.41 0.03a 0.48 0.00a 0.39 0.00a 0.30 0.06a 0.33 0.09a 0.35

C16:0 20.41 0.98a 20.90 0.10ab 21.40 0.03abc 2.50 0.11bc 21.84 0.35abc 22.03

C17:0 0.91 0.13a 1.89 0.01bc 1.89 0.00bc 1.83 0.01b 1.89 0.04bc 1.86

C18:0 3.97 0.12a 4.36 0.01ab 5.34 0.02bc 5.81 0.07c 5.11 0.45bc 4.43

C22:0 0.07 0.01a 0.05 0.00ab 0.05 0.00b 0.00 0.00c 0.04 0.00b 0.04

C23:0 0.11 0.01a 0.11 0.00a 0.10 0.00ab 0.08 0.00c 0.07 0.00c 0.07

SFA 33.30 1.42a 34.88 0.14ab 36.36 0.06bc 37.85 0.19cd 36.90 0.34bcd 36.34

C14:1n5 0.04 0.01a 0.04 0.00ab 0.03 0.00b 0.03 0.00b 0.03 0.00b 0.04

C16:1n7 9.83 0.51ab 9.25 0.05a 10.06 0.01ab 10.43 0.07b 10.68 0.29b 10.29

C17:1n9 0.95 0.17a 1.11 0.01ab 1.34 0.00c 1.25 0.01bc 1.33 0.03bc 1.23

C18:1n9 3.90 0.07b 3.48 0.01a 4.58 0.00c 5.24 0.03d 4.53 0.08c 4.51

C20:1n9 3.60 0.08a 2.84 0.01b 2.75 0.00b 2.31 0.02c 1.66 0.16d 1.66

C22:1n9 2.79 0.22a 2.17 0.01b 1.87 0.00c 1.18 0.01d 0.90 0.02e 0.72

C24:1n9 0.03 0.00a 0.03 0.00a 0.02 0.00bc 0.02 0.00cd 0.02 0.00bcd 0.03

6

4


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MUFA 21.12 0.40a 18.90 0.05b 20.65 0.01a 20.45 0.09a 19.18 0.48b 18.47 0.45

C18:2n6 1.84 0.03a 1.72 0.01b 1.50 0.00d 1.49 0.00d 1.49 0.01d 1.61 0.00

C18:3n6 0.33 0.02ab 0.37 0.00c 0.38 0.00c 0.36 0.00c 0.36 0.00bc 0.32 0.01

C18:3n3 0.79 0.01a 0.74 0.02b 0.55 0.00c 0.50 0.01d 0.47 0.00e 0.52 0.0

C20:2n6 0.14 0.00a 0.12 0.00b 0.10 0.00d 0.10 0.00cd 0.08 0.00e 0.10 0.0

C20:3n6 0.20 0.01a 0.21 0.01a 0.17 0.01b 0.20 0.00ab 0.17 0.02ab 0.19 0.0

C20:3n3 0.72 0.02a 0.71 0.07a 0.65 0.00ab 0.61 0.00b 0.65 0.02ab 0.72 0.0

C20:4n6 0.10 0.02a 0.03 0.01b 0.03 0.01b 0.06 0.01b 0.05 0.01b 0.05 0.0

C20:5n3 22.02 0.54a 21.77 0.03a 22.45 0.00a 22.14 0.08a 24.51 0.27b 23.91 0.2

C22:2n6 0.89 0.05a

0.93 0.01a

0.89 0.00a

0.87 0.01a

0.91 0.01a

0.88 0.0C22:5n3 4.96 0.34a 4.67 0.03a 5.06 0.01a 5.08 0.06a 5.14 0.12a 5.22 0.1

C22:6n3 13.61 0.92b 14.96 0.10a 11.22 0.03cd 10.27 0.11de 10.15 0.23de 11.79 0.2

PUFA 45.58 1.81ab 46.23 0.09a 42.99 0.06bc 41.69 0.27c 43.98 0.62abc 45.30 0.6

n-3 42.09 1.82a 42.85 0.10a 39.93 0.05ab 38.61 0.26b 40.92 0.63ab 42.15 0.6

Values are % mean S.D. of twenty-one homogeneous samples consisting of seven sardines from each size category (100-150 g, 150

Values in the same row labeled with a different superscript letter are significantly different (p 0.05).

65


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which is in agreement with several other studies (Bandarra et al., 1997;Beltran and Moral, 1991; Leonardis and Macciola, 2004; Shirai et al.,2002). The saturatedfatty acid (SFA) fraction increasedfrom33.30% to38.63% as the season progressed with 16:0 as the predominant fattyacid. Later in the season, there were significantly higher levels of 16:0(22.72%) compared to early season (20.41%), while other fatty acidssuch as 14:0 and 15:0 did not show significant differences as the seasonprogressed (p < 0.05).

The monounsaturated fatty acid (MUFA) fraction ranged from18.27% to 21.12%, with the predominant fatty acid being 16:1n7 fol-lowed by 18:1n9. Minor increases of these fatty acids were found as theseason progressed. On the other hand, the levels of 20:1n9 and 22:1n9

decreased from June to September (3.60% to 2.79% and 1.33% to0.61%, respectively). As the highest fatty acid fraction, PUFA fractionranged from 41.69% to 46.23% with the lowest concentration in mid-August and the highest in early July. The PUFA fraction consistedmainly of n-3 PUFAs (18:3n3, 20:3n3, 20:5n3, 22:5n3, and 22:6n3)with relatively high levels of 20:5n3 (range from 21.77% to 24.51%)throughout the season.

Sardines harvested off the OR-WA coast showed relatively higheramounts of n-3 PUFA levels compared with other species such asSardina pilchardus W caught in Spain (23.21% n-3 LC-PUFAs) andSardinops melanostictus caught in Japan (28.8% to 42.0% n-3 LC-PUFAs) (Beltran and Moral, 1991; Shirai et al., 2002). A similar spe-cies (Sardinops sagax caeruleus) harvested in Mexican waters in the

Gulf of California showed similar concentrations to our study of both20:5n3 and 22:6n3 ranging from 18.26% to 23.91% and 9.61% to13.70%, respectively (Gamez-Meza et al., 1999). Our study also showedhigher 20:5n3 levels than 22:6n3 throughout the season, and the sametendency was found by others (Bandarra et al., 1997; Kotb and AbuHadeed, 1991). However, some researchers reported higher 22:6n3 lev-els than 20:5n3 in sardine lipid, such as the Beltran and Moral study(1991) which showed 22:6n3 levels of 11.30% and 20:5n3 of 9.44%;Castrillon et al. (1997) reporting C22:6n3 levels of 16.92% and 20:5n3of 12.44%; and Leonardis and Macciola (2004) reporting 22:6n3 levelsof 11.3% and 20:5n3 of 6.5%. Researchers have suggested that the pro-portional changes of 20:5n3 and 22:6n3 can be influenced by the plank-ton species in their diet. In the post-spawning period when sardines start

to feed eagerly, 20:5n3 levels increase with a proportional decrease of22:6n3, as the predominant fatty acid in plankton is 20:5n3 (Bandarraet al., 1997).



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Minor increases of 20:5n3 levels were found as the season pro-gressed, from approximately 22% in June-August to 24% in August-September (p < 0.05). A similar tendency has been observed in otherstudies and it was suggested that seasonal blooms of phytoplankton andspawning cycles could be the cause for these changes (Gamez-Mezaet al., 1999). The relationship among 20:5n3 levels and 22:6n3, seasonand total lipid content was evaluated by multivariable linear regressionusing the model 20:5n3 = 1 (days) 2 (22:6n3) 3 (lipid).Results (R2 = 0.67,p < 0.001) showed a positive correlation betweenseason and 20:5n3 content (1 = 0.03,p < 0.001) at constant 22:6n3 andlipid content, showing an increase in 20:5n3 was associated with a sta-tistically significant progression of season. In contrast, there were no

significant relationships between 20:5n3 and 22:6n3 (2 = 0.24, p


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The Effect of Sardine Size on Lipid Content

The relationship between percent lipid and sardine size over the pe-riod of July 1 to August 9 is shown in Figure 6. Sardines were separatedinto two size categories by weight (150.0-199.9 g and 200.0-249.9 g)with an average weight of 179.8 g and 220.81 g, while lipid and mois-ture content were determined separately. There was a significant differ-ence found in lipid content between these two size groups (p < 0.05).The highest difference in lipid content was found in July, with the200.0-249.9 g size group having higher lipid levels than 150.0-199.9 gsize group.

CONCLUSIONS

This study showed seasonal differences in sardine compositionduring the 2004 harvest season off the OR-WA coasts. With propor-tional decrease of moisture content, sardine lipid content significantlyincreased from June to August and slightly decreased in September.The high lipid content resulting in high n-3 LC-PUFA levels in sardinessuggests that sardines captured in August-September have higher


50

60

55

50

40

30

20

10

07/1 7/6

lipid (150-199.9 g sample) lipid (200-249.9 g sample)

moisture (150-199.9 g sample) moisture (200-249.9 g sample)

7/8 7/12

Lipid(%)

7/15 7/19 7/26 8/2 8/940

45

Moisture(%)

Date

FIGURE 6. Effect of sardine size on lipid and moisture content.


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potential for oil recovery and should be preferred as raw material forhuman consumption or development of nutraceuticals. The consump-tion of 100 g of sardine provides higher levelsof n-3 LC-PUFAs, in par-ticular 20:5n3 and 22:6n3 fatty acids, than human daily requirements.

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Articel of Okada Et Morisey (2007)