ARTICLE IN PRESS

LWT 41 (2008) 483492 www.elsevier.com/locate/lwt

Stability of potassium iodide and omega-3 fatty acids in fortied freshwater sh emulsion sausage
Worawan Panpipata, Jirawat Yongsawatdigulb,
b

School of Food Technology, Institute of Agricultural Technology, Walailak University, Nakhonsithammarat 80160, Thailand School of Food Technology, Institute of Agricultural Technology, Suranaree University of Technology, Nakhon Ratchasima 30000, Thailand Received 9 September 2006; received in revised form 2 December 2006; accepted 20 March 2007

Abstract Oxidative stability of emulsion sausages prepared from African walking catsh (AF: Clarias gariepinus) and rohu (RH: Labeo rohita) fortied with three levels of the rened tuna oil (2%, 6%, and 10%) with 150 mg KI/100 g sample and without KI was investigated. The control was prepared using 20% soybean oil and without KI. Samples were vacuum-packed and stored at 4 1C. Iodine content decreased approximately 14% after cooking and remained constant throughout storage. Sausages fortied with tuna oil showed higher level of n-3 (eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)), but lower level of n-6 fatty acids (Po0.05) than the control. The ratio of n-6 to n-3 fatty acids of all samples decreased with an increase of tuna oil addition and was stable throughout 4 weeks of storage. Degradation of linoleic (LA) and linolenic acid (LNA) mainly occurred in the samples added 2% tuna oil whereas samples fortied with higher tuna oil (610%) exhibited higher loss of EPA and DHA. Hydroperoxide (HPV) and thiobarbituric reactive substances (TBARS) values were increased as addition level of tuna oil increased. Textural properties were not affected during storage at 4 1C for up to 4 weeks (Po0.05). Based on lipid stability results, 2% fortication of the rened tuna oil was recommended for emulsion sausage prepared from both species. Addition of 150 mg KI/100 g had no effect on lipid oxidation of sh sausages (Po0.05). r 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.
Keywords: Catsh; Rohu; Iodine stability; Lipid oxidation; Fish oil fortication

1. Introduction Iodine is an essential trace element required for synthesis of thyroid hormone, triodothyronine (T3) and thyroxine (T4). Thyroid hormones regulate a variety of physiological processes, including growth, metabolism, and reproductive function. Adolescents and adults need iodine in the amount of 150 mg/day (Muller & Krawinkel, 2005). Globally, 740 million people suffer from iodine deciency, including up to 300 million with goiter and 20 million with brain damage from maternal iodine deciency during pregnancy (Muller & Krawinkel, 2005). Iodine content of freshwater sh llet was about 37.5 mg/100 g meat, which was about 510 times lower than that of marine sh (Eckhoff & Maage, 1997; Haldimann, Alt, Blanc, & Blondeau, 2005). Nutritional
Corresponding author. Tel.: +66 44 224 359; fax: +66 44 224 150.

E-mail address: jirawat@sut.ac.th (J. Yongsawatdigul).

quality of processed food made from freshwater sh could be improved with the fortication of iodine. Marine sh are a rich source of n-3 polyunsaturated fatty acids (PUFA) containing approximately 1430% of total fatty acids, whereas PUFA in freshwater sh was only 111% of total fatty acids (Rahman, Huah, Hassan, & Daud, 1995; Steffens, 1997). Biological importance of n-3 PUFA, particularly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), on brain and retina development has been realized (Simopoulos, 1997). An increase in consumption of n-3 PUFA has been reported to reduce the risk of coronary heart disease, decrease mild hypertension, and prevent certain cardiac arrhythmias (Garg, Wood, Singh, & Moughan, 2006). Several foods including bread, milk and ice cream are successfully enriched with n-3 PUFA using either rened or encapsulated sh oil or n-3 PUFA from microalgae (Kolanowski, Swiderski, & Berger, 1999).

0023-6438/$30.00 r 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved. doi:10.1016/j.lwt.2007.03.013

ARTICLE IN PRESS
484 W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492

Emulsion sausages, such as frankfurter, are widely consumed in both Western and Asian countries. A product is typically made of beef, or beef and pork, or chicken and contains the fat content of 2530%. Fish mince and surimi have recently been used as a raw material for emulsion sausage production, particularly in Asian countries (Konno, 2005). Fortication of 3-n PUFA to the sh sausage could be an alternative means to improve its fat quality and to increase n-3 PUFA consumption. However, one of the main problems facing the application of marine sh oil in foods is their high susceptibility to oxidation. The oxidative deterioration involves the formation of hydroperoxides from polyunsaturated fatty acids. The progress of autooxidation gives rise to a complex mixture of secondary oxidation products. Although lipid hydroperoxide are tasteless and odourless, the secondary oxidation products are responsible for the changes in aroma and avour (Frankel, 1991). Oxidative deterioration is also related to the adverse changes in texture, appearance, and nutritional value (Min & Boff, 2002). In addition, some research works indicated the effect of iodine on lipid oxidation in biological systems. Verditti, Deleo, and Dimeo (1997) indicated that lipid peroxidation could be generated in patients with hyperthyroidism (excess iodine for body need). Similarly, iodine increased lipid peroxidation in goiter rodent (Allen, 1996). Thus, addition of iodine and tuna oil to sh emulsion sausage could have an effect on oxidative stability of the product. African walking catsh (AF, Clarias gariepinus) and rohu (RH, Labeo rohita) are abundant freshwater sh in Thailand and other Asian countries, such as India and China (FAO, 2005). These species are not fully utilized with relatively low value in spite of their relatively good gel-forming ability. Good textural properties of sh sausage can be obtained using these species as a raw material. The objective of this study was to investigate the effect of KI and the rened tuna oil fortication on oxidative stability of emulsion sausages prepared from AF and RH minces.

Emulsion sausages were prepared with the similar process to frankfurter production, using 60% sh mince and 20% soybean oil. In the treatment samples, soybean oil was substituted with the commercial rened tuna oil at 10%, 30%, and 50% of total oil addition, which was equivalent to 2%, 6%, and 10% of the total weight, respectively. The rened oil was prepared from tuna byproducts. Potassium iodide (KI) was added at either 0 or 150 mg/100 g sample. The controls were prepared using 20% soybean oil without KI. Minces were chopped using a Stephan vacuum cutter (UM5, Stephan Machinery Co., Columbus, OH, USA). Sodium chloride was added at 2% of total weight, subsequently soybean and tuna oil was added. Moisture content of all samples was adjusted to 86%. Other ingredients (0.2% tripolyphosphate, 1.6% spices, 2.4% tapioca starch, 1% soy protein isolate, and 1% sugar) were added. The raw paste was stuffed into a 22mm-diameter cellophane casing and pre-incubated at 55 1C for 40 min prior to cooking at 80 1C for 15 min. Smoking process was omitted in this study in order to reduce the colour interference of smoke compounds on thiobarbituric reactive substances (TBARS) determination. Samples were cooled for about 30 min in iced water, vacuum-packed and stored at 4 1C. Samples were randomly selected at 0, 1, 2, 3, and 4 weeks of storage for chemical and textural analyses. 2.2. Fatty acid proles Fatty acid proles of sh tissue and sausage samples during storage were determined using gas chromatography (GC) according to the AOAC method (1997). Total lipids were extracted according to the modied Folchs method (Folch, Less, & Stanley, 1957). The fatty acid methyl esters (FAME) were prepared by the addition of 1.5 ml of 0.5 M sodium hydroxide in methanol to 25 mg of lipid. The mixture was heated in boiling water bath (100 1C) for 2 min. After cooling, 2 ml of 14% boron triuoride in methanol was added in the mixture and reheated in a boiling water bath for 30 min. Subsequently, 1 ml of isooctane was added. Five millilitres of saturated NaCl was added to the mixture to separate isooctane phase (containing FAME) from methanol/water phase. Isooctane fraction was collected and the isooctane extraction was repeated once more. Two millilitres of isooctane (containing FAME) was dried under nitrogen gas at room temperature. Subsequently, 1 ml of isooctane was added to solubilize the dried FAME. The FAME was analysed using GC (6890 GC, Agilent Technology Co. Ltd., Palo Alto, CA, USA), equipped with a ame ionization detector. Fused silica capillary column (Supelcowax 10 column, SUPELCO Co., Ltd., Bellefonete, PA, USA) was used. Temperature was programmed from 140 to 240 1C at a rate of 4.0 1C/min. Temperature of both injector and detector was set at 260 1C. Carrier gas was helium and a ow rate of 0.9 ml/min was maintained throughout the experiment. Identication of FAME was based on comparison of retention time of tested samples to methyl

2. Materials and methods 2.1. Sample preparation African walking catsh (AF, C. gariepinus) and rohu (RH, L. rohita) were obtained from the sh farm at Nakhon Ratchasima. Live sh were transported to the Suranaree University of Technology Laboratory and immediately sacried upon arrival. The average AF and RH weights were 1650735.40 and 170.1672.5 g, respectively, with the lengths of 52.2476.12 and 24.571.60 cm, respectively. Fish were headed, degutted, and washed with cold water. Skin and bones were manually removed. Fish llets were ground using a mincer (Biro 8-22, Biro Manufacturing, Marblehead, OH, USA) with a screen size of 1.5 mm perforation.

ARTICLE IN PRESS
W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492 485

ester standards from Sigma (St. Louise, MO, USA). Absolute response factors were calculated for each identied fatty acid peak. Fatty acid composition was expressed as mg/100 g sample and was determined in duplicate. The ratio of n-6 to n-3 of sausage samples was calculated. The n-6 fatty acids used in the calculation were linoleic (LA), eicosatrienoic, and archidonic acid. Linolenic (LNA), eicosapentaenoic (EPA), and docosahexaenoic (DHA) were used for calculation of n-3 fatty acids. 2.3. Iodine content Iodine content of sh tissue and sausage samples during storage was measured by a spectrophotometric method according to Moxon and Dixon (1980). Fish mince was charred with 30% potassium carbonate solution at 550 1C for 3 h to eliminate all organic matters. Subsequently, the cooled ash was transferred to centrifuge tube with 50 ml of deionized (DI) water. The samples were then mixed with 0.023% potassium thiocyanate solution and 0.077% ammonium iron(III) sulfate. At exactly 90 s intervals, 2.07% sodium nitrite solution was added. The absorbance of mixture was measured at 450 nm after 20 min incubation. Iodine content was calculated using KI as a standard. 2.4. Thiobarbituric reactive substances (TBARS) Changes of TBARS were performed as described by Ahn, Lutz, Cherian, Wolfe, and Sim (1995). Ten grams of sample was mixed with 50 ml of 3.86% perchloric acid and butylated hydroxyanisole (BHA) to contain 125 mg/mg fat using a homogenizer (Nissei AM-8, NSS Nihonseiki Kaisha Co., Ltd., Kyoto, Japan) at high speed for 30 s. The homogenate was then ltered through a Whatman no. 1 lter paper, and a 2-ml aliquot of the ltrate was mixed with 2 ml 0.02 M thiobarbituric acid (TBA). Samples were heated in boiling water for 30 min. The absorbance was determined at 531 nm. DI water was used as a blank. 1,1,3,3-Tetraethyloxypropane (TEP) was used as a standard. TBARS was expressed as mg malonaldehyde/100 g sample. 2.5. Hydroperoxide value (HPV) Changes of HPV during storage were monitored using a spectrophotometic method as described by Shantha and Decker (1994). One gram of sample was homogenized with 10 ml of chloroform/methanol (2/1, v/v). After centrifugation at 2000 g for 5 min, an aliquot (2 ml) of the lower chloroform layer was mixed with an additional 1.3 ml of chloroform/methanol (2/1, v/v) and then reacted with 50 ml of 3.94 M ammonium thiocyanate and 0.144 M ferrous iron(II) solution. The reaction mixture was incubated at ambient (28 1C) for 5 min and absorbance at 500 nm was measured. Hydroperoxides oxidize Fe2+ to Fe3+ which subsequently formed complex with ammonium thiocyanate, resulting in pink colour. Hydroperoxide was quanti-

ed using a standard ferric iron(III) chloride prepared at various concentrations ranging from 0 to 40 mg/ml.

2.6. Textural properties Texture Analyzer (Stable Micro System, Surrey, UK) was used to evaluate changes of textural properties of sausages during storage. Samples were cut into pieces of 2 cm length. Breaking force (g) and deformation (mm) were determined using a 5-mm spherical plunger probe at a test speed of 1 mm/s.

2.7. Statistical analysis Two different lots (replications) of sh were used for the entire study. In chemical analyses, measurements were conducted at least in duplicate for each lot of sh. At least 10 specimens were used for textural measurement for each replication. Data from two replications were pooled and mean values7standard deviations were presented. The effects of iodine and tuna oil fortication on iodine stability and lipid oxidation were analysed by analysis of variance (ANOVA) using the Statistical Analysis System (SAS Institute Inc., Carry, NC) as a factorial design in randomized complete block design (RCBD). Signicant difference of means were determined using the Duncans multiple range test (DMRT). Level of signicance was set at Pp0.05.

Table 1 Fatty acid proles of the rened tuna oil and extracted lipid from African catsh (AF) and rohu (RH) minces Fatty acid Content (mg/100 g oil) AF Myristic acid (C14:0) Pentadecanoic acid (C15:0) Palmitic acid (C16:0) Palmitoleic acid (C16:1) Stearic acid (C18:0) Oleic acid (C18:1n9c) Linoleic acid (C18:2n6c) Arachidic acid (C20:0) Eicosenoic acid (C20:1) Octatrienoic acid (C18:3n6) Linolenic acid (C18:3n3) Eicosatrienonic acid (C20:3n6) Arachidonic acid (C20:4n6) Eicosapentaenoic acid (C20:5n3) Docosahexaenoic acid (C22:6n3) n-6 fatty acids n-3 fatty acids n-6/n-3 ratio 12.92 3.13 652.18 100.67 196.35 980.07 821.41 4.02 10.26 94.65 38.47 38.83 59.77 1.19 99.07 1014.66 138.73 7.31 RH 22.31 7.59 148.02 41.04 54.64 200.20 60.20 1.88 2.93 2.62 57.83 5.88 86.35 9.59 256.22 155.05 323.64 0.48 Tuna oil 2645.62 863.79 14251.20 5045.58 4255.87 8532.93 1290.22 329.75 437.28 449.50 391.98 108.17 82.46 4023.14 26680.54 1930.36 31095.65 0.06

ARTICLE IN PRESS
486 W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492

3. Results and discussion 3.1. Iodine content and fatty acid composition of sh tissue RH mince contained lipid content of 1.7670.02%, while lipid content of AF was 9.1070.1%. Iodine content of AF and RH mince was 12.6770.3 mg/100 g and 12.8370.1 mg/100 g, respectively. Eckhoff and Maage (1997) found that iodine content of catsh esh in the

East and West Greenland was 7.8 mg/100 g meat. Some variations in iodine content were inuenced by age, diet, and habitat of sh. Hunt and Eales (1979) found that 84% of plasma iodine in rainbow trout came from water, 16% from the diet, and less than 1% from thyroid hormone degradation. Iodine content of marine sh is typically higher than that of freshwater sh and has been reported to vary from 50 to 560 mg/100 g meat (Eckhoff & Maage, 1997).

Table 2 Iodine stability of African catsh (AF) and rohu (RH) sausages fortied with 150 mg KI/100 g and various levels of tuna oil during storage at 4 1C Fish Tuna oil substitution (%) Storage time (week) 0 AF 0 2 6 10 0 2 6 10 130.0570.19 131.4570.31 131.8570.22 130.6570.45 132.2570.47 129.6570.56 131.6770.23 130.2470.15 1 130.5670.26 130.4270.11 131.2570.52 131.3570.61 131.8670.17 130.2570.75 130.2570.32 131.2570.46 2 130.4670.18 130.2570.05 130.2870.50 130.8770.16 130.2470.25 130.5870.19 130.4570.23 130.2570.57 3 130.2570.28 130.4570.31 130.8770.14 130.8670.42 130.4570.43 130.2870.37 131.2470.16 130.5470.24 4 129.7870.27 129.1570.34 129.5470.15 130.1570.24 129.2470.58 129.4570.64 130.4570.24 129.2670.26

RH

LA 12000 Content (mg/100g sample) Content (mg/100 g sample) 10000 8000 6000 4000 2000 0 0 1 3 2 Storage time (wk) LNA 1200 Content (mg/100 g sample) Content (mg/100 g sample) 1000 800 600 400 200 0 0 1 2 3 Storage time (wk) 0% FO 4 5 1200 1000 800 600 400 200 0 0 1 4 5 12000 10000 8000 6000 4000 2000 0 0 1

LA

2 3 Storage time (wk) LNA

2 3 Storage time (wk)

2% FO

6% FO

10% FO

Fig. 1. Changes of some PUFA content of AF (a, c, e and g) and RH (b, d, f and h) emulsion sausages fortied with 150 mg KI/100 g sample and various levels of tuna oil, stored at 4 1C. LA: linoleic acid, LNA: linolenic acid, EPA: eicosapentaenoic acid, DHA: docosahexaenoic acid, FO: sh oil.

ARTICLE IN PRESS
W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492 487

EPA 500 Content (mg/100 g sample) 400 300 200 100 0 0 1 2 3 Storage time (wk) DHA 3000 Content(mg/100 g sample) Content (mg/100 g sample) 2500 2000 1500 1000 500 0 0 1 3 2 Storage time (wk) 0% FO 4 5 3000 2500 2000 1500 1000 500 0 0 1 4 5 Content (mg/100 g sample) 500 400 300 200 100 0 0 1

EPA

2 3 Storage time (wk) DHA

2 3 Storage time (wk)

2% FO

6% FO

10% FO

Fig. 1. (Continued)

AF muscle contained higher saturated fatty acids (palmitic and stearic acid) and LA content than did RH muscle (Po0.05, Table 1). Total n-3 PUFA content of AF was lower than that of RH (Po0.05, Table 1), and much lower than those of salmon (10001400 mg/100 g) and trout (5001600 mg/100 g) (Sidhu, 2003). Healthy diet should contain high amount of n-3 PUFA and lower n-6 PUFA. High amount of n-6 PUFA in diets promoted the pathogenesis of several diseases, including cardiovascular disease, cancer, and inammatory as well as autoimmune diseases (Simopoulos, 2002). Therefore, the lower ratio of n-6 to n-3 PUFA is more desirable as far as the health benets are concerned. Garg et al. (2006) indicated that the optimal ratio of n-6 to n-3 PUFA should not exceed 41. Thus, proportion of PUFA content of RH esh was in the optimal ratio (Table 1). However, it might not be a good source of n-3 PUFA due to its lower lipid content. Moreira, Visentainer, de Souza, and Matsushita (2001) found that the ratio of n-6/n-3 of wild freshwater sh was 1.141.79, which was higher than that of marine sh (0.050.08). The commercial tuna oil contained low amount of n-6 PUFA but high level of n-3 PUFA content (Table 1). Fortication of tuna oil into a product made

from AF and RH could improve fatty acid composition to attain the optimum ratio for health benets. 3.2. Iodine stability of sh sausage Although KI at the concentration of 150 mg/100 g was added, the nal iodine content of the cooked emulsion sausages were reduced to approximately 130 mg KI/100 g, reecting about 1416% loss after cooking (Table 2). Iodine content remained constant throughout 4 weeks of storage (Table 2). It should be noted that iodine content of sausages without KI fortication also decreased about 1520% (data not shown). The loss of iodine might be due to the exposure to high temperature during sausage cooking. Diosady, Alberti, and Mannar (2002) indicated that the critical problem of iodine stability is the potential for reducing or oxidizing the iodide to elemental iodine, I2, which readily sublimes and is lost through diffusion at room temperature, or 440 1C. High stability of iodine up to 4 weeks was attributed to low temperature storage (4 1C). Chauhan, Bhatt, and Majeethai (1992) reported that iodine loss of 910% occurred in a common iodized salt within the rst month of storage.

ARTICLE IN PRESS
488 W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492 Table 3 Changes of n-6/n-3 ratio of African catsh (AF) and rohu (RH) sausages fortied by KI and various levels of tuna oil during storage at 4 1C Fish Soybean Fish Added Storage time (week) oil (%) oil (%) iodine level 1 2 3 (mg KI/100g 0 sample) AF 20 18 14 10 20 18 14 10 RH 20 18 14 10 20 18 14 10 0 2 6 10 0 2 6 10 0 2 6 10 0 2 6 10 0 9.97 6.21 3.20 1.72 9.72 6.68 3.09 1.56 8.86 5.75 2.61 1.25 9.06 5.38 2.59 1.26 9.61 6.87 3.19 1.79

3.3. Changes of PUFA during storage Lipid content of sausages made from AF mince was approximately 25%, while that from RH mince was about 20.6%. LA and LNA content were high in all samples because soybean oil was used in the formulation (Fig. 1). Addition of tuna oil increased EPA and DHA content and decreased LA and LNA content of both AF and RH sausages (Fig. 1). The average content of DHA of sh sausages fortied with 2% tuna oil was 549.2 mg/100 g, which was comparable to the content of naturally occurring DHA found in wild salmon esh (Bell et al., 1998). Fortication of 610% tuna oil reduced the ratio of n-6/n-3 to be less than 4 (Table 3), which was considered to be more desirable for health benets. A lower ratio of n-6/ n-3 (23) in the diets has been reported to reduce rectal cell proliferation in patients with colorectal cancer, suppress inammation in patients with rheumatoid arthritis and decrease the risk or coronary heart diseases (Simopoulos, 2002). Kouba, Enser, Whittington, Nute, and Wood (2003) reported that feeding a diet containing 6% whole crushed linseed (rich source of n-3 fatty acids) reduced the n-6/n-3 ratio in longissimus muscle from 7.6 to 3.9. Nutritional

9.43 10.12 10.04 6.30 6.16 6.14 3.17 3.00 3.03 1.69 1.64 1.67 9.62 6.23 2.97 1.69 9.03 5.40 2.50 1.30 8.77 5.39 4.14 1.27 9.51 6.05 2.98 1.75 9.34 5.44 3.05 1.32 8.80 5.62 2.72 1.25

150

9.43 10.70 6.19 6.30 3.13 3.11 1.77 1.85 8.19 5.34 2.87 1.20 9.97 4.71 2.65 1.21 8.87 5.53 2.68 1.24 8.85 5.43 2.52 1.23

150

No KI Addition 40 35 HPV (mg/kg sample) HPV (mg/kg sample) 30 25 20 15 10 5 0 0 1 3 2 Storage time (wk) 4 5 40 35 30 25 20 15 10 5 0 0 1

150 g KI/ 100 g sample

2 3 Storage time (wk)

60 50 HPV (mg/kg sample) 40 30 20 10 0 0 1 3 2 Storage time (wk) 0% FO 4 5 HPV (mg/kg sample)

60 50 40 30 20 10 0 0 1 3 2 Storage time (wk) 4 5

2% FO

6% FO

10% FO

Fig. 2. Changes of HPV of AF (a, b) and RH (c, d) sausages stored at 4 1C; (a, c) without KI addition; (b, d) 150 mg KI/100 g sample, FO: sh oil.

ARTICLE IN PRESS
W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492 489

quality, especially PUFA composition and ratio, of sh sausage was improved through substitution of 6% rened tuna oil (Fig. 1 and Table 3). However, shy note was observed in samples substituted with 46% sh oil. Several volatile compounds contributed to shy odour of sh oil, including pentylfuran, decatrienal, and heptenal (PerezMateos, Boyd, & Lanier, 2004). Sensory characteristic of the product should be one of important factors taken into consideration in selecting the optimal level for sh oil addition. It should be mentioned that KI fortication had no effect on stability of individual fatty acids during storage of 4 weeks. LA and LNA content of the control (20% soybean oil) and sample added 2% tuna oil (18% soybean oil) dramatically decreased after 3 weeks of storage (Fig. 1). However, EPA and DHA of these samples remained constant throughout the studied period. These results indicated that LA and LNA primarily underwent oxidation in samples containing low level of PUFA (control and 2% tuna oil substitution). DHA and EPA of samples added 6% and 10% tuna oil decreased after 3 weeks of storage (Po0.05). A decrease of EPA and DHA appeared to be greater than that of LA and LNA. EPA and DHA contain

higher degree of unsaturation, thus are more susceptible to oxidation. In general, approximately 1315% loss of DHA and EPA was found in sausages substituted with 610% tuna oil at the 4th week of storage. The greatest DHA loss of 32.9% was observed in the RH samples fortied with 6% tuna oil and KI (Fig. 1h). It should be noted that the n6/n-3 ratio remained constant throughout storage time (Table 3). These results indicated that oxidation of n-6 occurred at the same extent as that of n-3 during storage, regardless of the sh oil substitution level. 3.4. Changes of HPV and TBARS values HPV and TBARS values of samples prepared from AF and RH increased when substitution level of tuna oil increased (Po0.05) (Figs. 2 and 3, respectively). This was due to higher content of unsaturated fatty acids, especially EPA and DHA. HPV and TBARS gradually increased with storage time. Lin, Lin, and Kuo (2002) reported that frankfurters made from chicken fed diet enriched with 2% and 4% supplemental sh oil showed comparable TBARS to that of the control. Less extent of oxidation was probably due to the antioxidative activity of added nitrite.

No KI addition 8 7 TBARS (mg /kg sample) 6 5 4 3 2 1 0 0 1 3 2 Storage time (wk) 4 5 TBARS (mg/kg sample) 8 7 6 5 4 3 2 1 0 0 1

150g KI/100 g sample

3 2 Storage time (wk)

14 TBARS (mg/kg sample) TBARS (mg/kg sample) 1 2 3 Storage time (wk) 0% FO 4 5 12 10 8 6 4 2 0 0

14 12 10 8 6 4 2 0 0 1 3 2 Storage time (wk) 4 5

2% FO

6% FO

10% FO

Fig. 3. Changes of TBARS of AF (a, b) and RH (c, d) sausages stored at 4 1C; (a, c) without KI addition; (b, d) 150 mg KI/100 g sample, FO: sh oil.

ARTICLE IN PRESS
490 W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492

Addition of 1.52.5% menhaden oil or puried marine oil also did not promote lipid oxidation of surimi seafood kept at 4 1C for 2 months, resulting in small amount of oxidative products and subtle change in sensory characteristics (Perez-Mateos et al., 2004). Our results demonstrated that addition of the rened tuna oil to emulsion sh sausage appeared to accelerate lipid oxidation, especially at the level of 610% substitution. In general, lipid stability of samples fortied with 2% tuna oil was comparable to that of the control (Figs. 2 and 3). Therefore, addition of the rened tuna oil should be controlled at 2%. To maximize the level of fortication possible (6%) without leading to lipid oxidative instability, microencapsulated sh oil in conjunction with a proper antioxidant and/or chelating agents should be applied. KI had no effect on HPV and TBARS values of all sample (P40.05). Therefore, it can be fortied at the level of 150 mg/100 g sample without promoting lipid oxidation. It should be noted that HPV and TBARS values of sausages made from RH were higher (Po0.05) than those made from AF in spite of the comparable PUFA content of the nished products at day 0 (Fig. 1). Aubourg et al.

(2005) reported that lipid oxidation of coho salmon occurred at a slower rate than other fatty sh due to the presence of natural free radical scavenger, astaxanthin. RH is typically classied as a lean sh and contains lower lipid content than AF. Differences in intrinsic properties, such as the amount of dark muscle, metal ions, membrane lipids, and natural antioxidant in the muscle, could contribute to the different degree of lipid oxidation between these two species. 3.5. Textural properties Fortication of the rened tuna oil had no effect on textural properties of both AF and RH sausages (P40.05, Fig. 4ad). Breaking force and deformation remained relatively constant during storage (P40.05). Addition of KI did not affect textural properties of sh sausage (data not shown). Perez-Mateos et al. (2004) also reported that fortication of either 1.5% or 2.5% sh oil had no effect on textural properties of surimi seafoods during 2 months of storage. Breaking force values of RH sausage were lower than those of AF, indicating that gel-forming ability of RH

320 280 Breaking force (g) Deformation (mm) 240 200 160 120 80 0 1 2 3 Storage time (wk) 4 5

11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 0 1 3 2 Storage time (wk) 4 5

320 280 240 200 160 120 80 0 1 2 3 Storage time (wk) 0% FO 4 5 Deformation (mm) Breaking force (g)

11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 0 1 2 3 Storage time (wk) 4 5

2% FO

6% FO

10% FO

Fig. 4. Changes of breaking force (a, c) and deformation (b, d) of AF (a, b) and RH (c, d) sausages fortied with various levels of tuna oil and stored at 4 1C, FO: sh oil.

ARTICLE IN PRESS
W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492 491

muscle was inferior to that of AF muscle. Differences in intrinsic characteristics, such as endogenous proteinases and transglutaminase activity, between two species could contribute to different textural properties and deserve further investigation. A substance generated from lipid oxidation, malonaldehyde, has been reported to decrease gel forming ability of myobrillar proteins (Liu & Xiong, 2000; Liu, Xiong, & Butterled, 2000). Malonaldyhyde decreased stability of myosin conformation, increased protein carbonyls, and promoted myosin heavy chains cross-linking. As a result, gel forming ability of myosin heavy chain decreased. In our system, however, protein gel networks were formed before the onset of lipid oxidation. Therefore, the effect of lipid oxidative products on the formation of protein networks in this case was minimal. 4. Conclusions AF and RH esh contained low level of iodine and n-3 fatty acids. Fortication of KI (150 mg/100 g sample) and rened tuna oil (2%, 6%, and 10%) had no effect on textural properties of sh sausages during storage at 4 1C. Addition of KI at the level of 150 mg/100 g samples did not promote lipid oxidation of sh sausage. Lipid oxidation increased with added level of tuna oil. Although addition of 6% and 10% tuna oil lowered the ratio of n-6 to n-3 to the optimal range for health benets, strong shy avour was noted in the products and lipid oxidation occurred to a greater extent during storage. Based on lipid stability results, 2% fortication of the rened tuna oil was recommended for emulsion sausage made from AF and RH. Acknowledgement This study was nancially supported by the National Center for Genetic Engineering and Biotechnology (BIOTEC), Thailand, under a research Grant BT-B-06-FG-1945-03. References
Ahn, D. U., Lutz, S., Cherian, G., Wolfe, F. A., & Sim, J. S. (1995). Lipid oxidation and sensory characteristics of fresh and cured sausage from linolenic acid enrich pork. Journal of Food Quality, 18, 397413. Allen, E. M. (1996). Acute iodine ingestion increases interphyroidal glutathione (online). Available: /http://www.glutathione.htmS. AOAC. (1997). Ofcial methods of analysis of the association of ofcial analytical chemists (15th ed.). Arlington: AOAC. Aubourg, S. P., Vinagre, J., Rodriguez, A., Losada, V., Larrain, M. A., Quitra, V., Gomez, J., Maier, L., & Wittig, E. (2005). Rancidity development during the chilled storage of farmed Coho salmon (Oncorhynchus kisutch). European Journal of Lipid Science and Technology, 107, 411417. Bell, J. G., McEvoy, J., Webster, J. L., McGhee, F., Mullar, R. M., & Sargent, J. R. (1998). Flesh lipid and carotenoid composition of Scottish farmed Atlantic salmon (Salmo salar). Journal of Agricultural and Food Chemistry, 46, 119127.

Chauhan, S. A., Bhatt, A. M., & Majeethai, K. M. (1992). Stability of iodized salt with respect to iodine content. Research and Industry, 37, 3841. Diosady, L. L., Alberti, J. O., & Mannar, M. G. V. (2002). Microencapsulation for iodine stability in salt fortied with ferrous fumarate and potassium iodide. Food Research International, 35, 635642. Eckhoff, K. M., & Maage, A. (1997). Iodine content in sh and other food products form East Africa analyzed by ICP-MS. Journal of Food Composition and Analysis, 10, 270282. FAO. (2005). Summary tables on capture: Aquiculture and trade. (online). Available: /http://www.fao.orgS. Folch, J., Less, M., & Stanley, S. G. H. (1957). A simple method for the isolation and purication of total lipids from animal tissues. Journal of Biological Chemistry, 226, 497509. Frankel, E. N. (1991). Review: Recent advances in lipid oxidation. Journal of the Science of Food and Agriculture, 54, 495511. Garg, M. L., Wood, L. G., Singh, H., & Moughan, P. J. (2006). Means of delivering recommended levels of long chain n-3 polyunstaturated fatty acids in human diets. Journal of Food Science, 71, R66R71. Haldimann, M., Alt, A., Blanc, A., & Blondeau, K. (2005). Iodine content of food groups. Journal of Food Composition, 18, 461471. Hunt, D. W. C., & Eales, J. G. (1979). Iodine balance in rainbow trout (Salmo gairdner) and effects of testosterone propionate. Journal of the Fisheries Research Board of Canada, 36, 282285. Kolanowski, W., Swiderski, F., & Berger, S. (1999). Possibilities of sh oil application for food products enrichment with o-3 PUFA. International Journal of Food Sciences and Nutrition, 50, 3949. Konno, K. (2005). New developments and trends in kababoko and related research in Japan. In J. W. Park (Ed.), Surimi and surimi seafood (2nd ed., pp. 847868). Boca Raton, FL: CRC Press, Taylor & Francis. Kouba, M., Enser, M., Whittington, F. M., Nute, G. R., & Wood, J. D. (2003). Effect of a high linolenic acid diet on lipogenic enzyme activities, fatty acid composition, and meat quality in the growing pig. Journal of Animal Science, 81, 19671979. Lin, J. H., Lin, Y. H., & Kuo, C. C. (2002). Effect of dietary sh oil on fatty acid composition, lipid oxidation and sensory property of chicken frankfurters during storage. Meat Science, 60, 161167. Liu, G., & Xiong, Y. L. (2000). Thermal transitions and gelation oxidative modied myosin, lactoglobulin, soy 7s globulin and their mixtures. Journal of the Science of Food and Agriculture, 80, 17281734. Liu, G., Xiong, Y. L., & Butterled, D. A. (2000). Chemical, physical, and gel-forming properties of oxidized myobrillar, whey, and soy protein isolates. Journal of Food Science, 65, 811818. Min, D. B., & Boff, J. M. (2002). Lipid oxidation of edible oil. In C. C. Akoh, & D. B. Min (Eds.), Food lipid chemistry, nutrition, and biotechnology (2nd ed., pp. 335363). New York: Marcel Dekker Inc. Moreira, A. B., Visentainer, J. V., de Souza, N. E., & Matsushita, M. (2001). Fatty acids prole and cholesterol contents of three Brazillian Brycon freshwater sh. Journal of Food Composition and Analysis, 14, 565574. Moxon, R. D. E., & Dixon, E. J. (1980). Semi-automatic method for determination of total iodine in food. Analyst, 105, 344352. Muller, O., & Krawinkel, M. (2005). Malnutrition and health in developing countries. Canadian Medical Association Journal, 173, 279286. Perez-Mateos, M., Boyd, L., & Lanier, T. (2004). Stability of omega 3 fatty acids in fortied surimi seafoods during chilled storage. Journal of Agricultural and Food Chemistry, 52, 79447949. Rahman, S. A., Huah, T. S., Hassan, O., & Daud, N. M. (1995). Fatty acid composition of some Malaysian freshwater sh. Food Chemistry, 54, 4549. Sidhu, K. S. (2003). Health benets and potential risks related to consumption of sh or sh oil. Regulatory Toxicology and Pharmacology, 38, 336344. Shantha, N. C., & Decker, E. A. (1994). Rapid, sensitive, iron-based spectrophotometric methods for determination of peroxide values of food lipid. Journal of AOAC International, 77, 421424.

ARTICLE IN PRESS
492 W. Panpipat, J. Yongsawatdigul / LWT 41 (2008) 483492 Steffens, W. (1997). Effects of variation in essential fatty acids on nutritive value of freshwater sh for humans. Aquaculture, 151, 97119. Verditti, P., Deleo, T., & Dimeo, S. (1997). Vitamin administration attenuates the tri-iodothyronine-induced of heart electrical activity in the rat. Journal of Experimental Biology, 200, 909914. Simopoulos, A. P. (1997). o-3 fatty acids in the prevention-management of cardiovascular disease. Canadian Journal of Physiology and Pharmacology, 75, 234239. Simopoulos, A. P. (2002). The importance of the ration of omega-6/ omega-3 essential fatty acids. Biomedicine and Pharmacotherapy, 56, 365379.