DETERMINATION OF SHELF LIFE OF FISH BALL MARINATED AFTER FRYING PROCESS

Abstract

The shelf life of fish ball marinated after frying process was investigated in the present study. The fish ball was stored at ±4°C. Total Volatile Basic Nitrogen (TVB-N), Thiobarbituric Acid Reactive Substances (TBAR), Trimethyl-Amine Nitrogen (TMA-N) and pH values of fish balls were 13.66 mg/100 g, 5.68 mg MA/kg, 5.63 mg/100 g and 3.42 at the end of the storage period (on day 150), respectively. The microbiological analysis results did not exceed the limit values. According to sensory evaluation results, the shelf life of balls was determined to be 135 days at ±4°C.

Headnote

ABSTRACT

The shelf life of fish ball marinated after frying process was investigated in the present study. The fish ball was stored at ±4°C. Total Volatile Basic Nitrogen (TVB-N), Thiobarbituric Acid Reactive Substances (TBAR), Trimethyl-Amine Nitrogen (TMA-N) and pH values of fish balls were 13.66 mg/100 g, 5.68 mg MA/kg, 5.63 mg/100 g and 3.42 at the end of the storage period (on day 150), respectively. The microbiological analysis results did not exceed the limit values. According to sensory evaluation results, the shelf life of balls was determined to be 135 days at ±4°C.

- Keywords: anchovy, fish ball, fried, marination, shelf life -

INTRODUCTION

Seafood may spoil sooner in contrast to other meat products due to higher water amount and lower connective tissue. (VARLIK et al., 2004). Under normal refrigerated storage conditions, the shelf life of these products is limited by enzymatic and microbiological spoilage (ASHIE et al, 1996). Microorganisms are the major cause of spoilage of most seafood products. However, only a few members of the microbial community like the specific spoilage organisms, give rise to the offensive off-flavours associated with seafood spoilage (GRAM and DALGAARD, 2002).

A great number of seafood processing techniques have been applied to slow down that spoilage. These processing technologies have been increased through developing knowledge. On the other hand, while that increase is being achieved, traditional methods have not been given up completely; in fact, these methods are still being used and developed (VARLIK et al, 2004).

Marination is one of the oldest processing methods that is used for preservation of fish and other seafood (GIUFFRIDA et al., 2007). Generally, marinated fish is ready to eat food that is not heatprocessed (GRAM and HUSS, 1996). Marination is the process of ripening fish and making it edible by treating with vinegar or organic acids and salt without using the heating process. Products after ripening process are packaged with brine, sauce, cream, mayonaisse or oil and served for consumption. The fresh, frozen and salted fish or fish parts may be used in marination technology. Fishes mainly used in marination are herring, anchovy, sardine, snakefish, trout, mackerel and silverside. Also, shellfish like mussel and shrimp may be processed as marinade. The best marinade product is made as a result of using fatty fish species like anchovy and herring. The marinated products can be split into 3 groups as cold, cooked and fried (GOKOGLU, 2002; BAYGAR et al., 2000).

1. Cold: The fresh material is being ripened in solution containing acetic acid and salt. There is no heat treatment.

2. Cooked: The fish are immersed into solution containing acetic acid and salt at 85°C. Thus, most of the bacterias are killed and enzymes are inactivated.

3. Fried: In material that is fried in acetic acid and salt solution before packaging, most of the bacteria are killed and enzymes are denaturated (CLUCAS and WARD, 1991; KILINC and CAKLI, 2004).

Fried marinade are products that are obtained by utilization of fried fish or fish products as marinade. For making fried marinade, fish is fried in vegetable oil previously and then immersed into solution (ERSAN, 1961; VARLIK et al., 1993). Fresh and frozen fish or fish parts are also fried and coated with brine or sauces. Herring, snakefish, river snakefish, whiting, codfish species, mediterranean sand smelt and some types of flatfishes are used to make fried marinade. The temperatures of frying in oil must be between 160° and 180°C. The frying time depends on temperature of oil, thickness and water content of the flesh of fish. The time of frying process is between 5-12 min. Rising of fried fish onto oil surface during frying process in fry-pan occurs as a result of their losing water and absorption of oil due to their specific weight. The fried fish must be packaged after it was cooled. The rate of fish: coating solution is approximately 2:1. This rate depends on absorption of solution by fried fish. The fish lose approximately 20% of their water during frying process. This loss is compensated from coating solution. The contents of acetic acid and salt of solution are 2-3.5% and 3-5%, respectively. But these rates can be changed depending on water content of product and seasonal changes (MEYER, 1965).

In this study, the fish ball produced with anchovy [Engraulis encrasicolus) was fried in oil and then marinated. It was aimed to determine shelf life of the fish ball marinated after frying.

MATERIALS AND METHODS

Materials

Anchovies [Engraulis encrasicolus, L.1758) were purchased from a fisherman in Sinop. In total, 10 kg of fresh anchovy with an average lenght of 9±1 cm were used. Fish were headed, gutted and washed.

Preparation of fish meat balls

The anchovies were boiled for 5 min and then minced with a blender after the bones were removed. The mixture was kneaded after addition of 0.58% semolina, 0.60% crumb, 0.83% egg, 1% parsley, 1% onion, 0.10% garlic, 0.08% salt, 0.03% black pepper, 0.03% cummin, 0.03% red pepper, 0.03% thyme and 0.03% ginger. Small pieces were taken apart from fish ball mix and given ball shape by hand. Then, they were fried in sunflower seed oil.

Marination process

The fried meat balls were put into the solution containing 7% salt and 1.5% vinegar after they were cooled. Besides, parsley, mustard seed, garlic and white pepper were added into the solution and jar caps were closed. Then, they were stored at 4°C for 150 days.

Chemical analysis

pH analysis was carried out with the instrument Werkstätten 82362 Weilheim, Germany, according to CURRAN et al (1980). Total vola- tile basic nitrogen (TVB-N) was determined according to method of Lucke and Geidel modified by ANTONACOPOULOS (1989). Thiobarbituric Acid Reactive Substances (TBAR) was determined according to TARLADGIS et al. (1960). TMA (mg TMA-N /100 g) analysis was carried out according to the method proposed by BOLAND and PAIGE (1971).

Microbiological analysis

Microbiological analyses were made according to Baumgart (19). 10 g of fish sample was taken, transferred into 90 mL sterile Physiological Saline Solution (0.85%) and then homogenized in a homogenizer (IKA Yellow Line DI 25 Basic). From the 10 1 dilution, other decimal dilutions were prepared and inoculated. Plate Count Agar was used as medium for total mesophilic aerobic bacteria and psychrophilic bacteria counts, petri dishes were incubated at 28°C for 3 days and 4°±1°C for 10 days, respectively. For total yeast-mold count, Potato Dextrose Agar was used as medium and petri dishes were incubated at 28°C for 3 days. To count coliform bacteria, Violet Red Bile Agar was used as medium and petri dishes were incubated at 35°C for 24 h. Results were given as log cfu/g. (ROGER et al, 1987; GOKTAN, 1990; VARLIK et al, 1993; GOKALP et al., 1999).

Sensory analysis

Sensory analyses were made according to NEUMAN et al. (1983). 10 g of fish sample was served to each panelist to evaluate the sensory attributes (appearance, odor, texture, flavor) of the sample. The samples were presented in separate plates to every panelist. According to the scoring test, a total score of sensory attributes of 20 was indicating excellent quality. Scores between 18.2 and 19.9 indicated "very good" quality, scores between 15.2 and 18.1 indicated "good" quality and scores between 11.2 and 15.1 indicated "middle" quality; scores between 7.2 and 11.1 indicated the limit of acceptability, and scores between 4.0 and 7.1 indicated spoiled samples.

Statistical analysis

The Minitab 15 (Minitab Inc. USA) program was used to search for significant differences among mean values of different results. Differences between means were analyzed by one - way analysis of variance (ANOVA). The results are presented as mean ± SE. The P value (P < 0.05) was used to determine significant differences.

RESULTS AND DISCUSSION

TVB-N values of the fish ball marinated after frying are shown in Fig. 1. The TVB-N content of fresh material was determined as 7.6 mg/100 g. The initial TVB-N content of fish ball marinated after frying was 5.6±0.00 mg/100 g. This value increased to 13.66±0.08 mg/100 gat the end of the storage period of 150 days. It was observed that TVB-N values increased significantly (P<0.05) during storage of the fish ball.

There have been limited number of studies related with the fish ball marinated after frying. However, there have been great number of studies about the marinades without frying (AKSU et al, 1997; DOKUZLU, 2000; GOKOGLU et al., 2004; OZDEN and BAYGAR, 2003; DUYAR and EKE, 2009; OZOGUL et al 2009; GUNSEN et al., 2011).

GOKOGLU et al. (2004) reported that TVB-N values of sardine marinade samples containing 2 and 4% acetic acid were 28.9 and 23.32 mg/100 g at the end of the storage period of 150 days, respectively. In another report TVBN N in anchovy marinated with acetic acid of 4% and stored at 4°C increased from 9.8 to 14 mg/100 g during the storage of 8 months (DOKUZLU, 2000). DUYARand EKE (2009) stated that TVB-N values of anchovy and bonito marinades were 17.63 and 18.67 mg/100 g at the end of the storage period of 170 days, respectively. These values were higher than our TVBN N (13.66 mg/100 g). Using different fish types and different rates of salt and acetic acid, application of heating process (frying) that has lethal effects in bacteria can be considered as the possible reasons of this situation. In addition, probability of solubilization in the marination solution of volatile amines might be effective to diminish TVB-N value. VARLIK et al. (2000) marinated fried anchovy meat ball similarly to our study and stored at 4°C. TVB-N val- ue of the product was reported as 10.45 mg/100 g at the end of the storage period of 150 days. This value was similar to the our finding (13.66 mg/100 g). Accordingly, it could be said that the frying process that was applied before marination was effective to diminish TVB-N value of marinated products.

One of the most important chemicals in fish muscle is trimethylamine N-oxide. Trimethylamine N-oxide is found in seafish mostly. TMA is produced by the decomposition of trimethylamine N-oxide caused by bacterial spoilage and enzymatic activity (SCHORMULLER, 1968; KIETZMANN, 1969). It was stated by FAO (1986) that, the TMA-N value is approximately 1 mg/100 g for fresh seafood and it is over 8 mg/100 g for spoiled samples. Besides, according to the TMAN N the seafood is classified as 'good' up to 4 mg/100 g; 'marketable' up to 10 mg/100 g; 'spoiled' if bigger than 12 mg/100 g(KUNDAKCI, 1989).

TMA values of the fish ball marinated after frying are shown in Fig. 3. The TMA content of the fresh material was 2.88 mg/100 g. In the present study, initial TMA-N value of the fish ball was found as 2.62 mg/100 g. At the end of the storage period of 150 days, it increased to 5.63 mg/100 g. In the study investigating the TMAN N of the anchovy ball marinated after frying, it was stated that the initial TMA-N value increased from 1.85 to 2.85 mg/100 gat the end of the storage period of 150 days (VARLIK et al, 2000). In our study, the TMA-N value was 2.88 mg/100 g initially and it was observed as 5.63 mg/100 gat the end of the storage. It was reported that the TMA content changes depending on season, catching area, species, type muscle and processing techniques (HERBARE) et al., 1982). It was thought that this difference was resulted from high initial TMA content and processing techniques such as immersing into different marination solution.

When compared with only marinated (without frying) samples, it was seen that our TMA values were higher than other studies. GOKOGLU et al. (2004) reported that TMA values of the sardine marinade samples containing 2 and 4% acetic acid were 1.67 and 0.86 mg/100 g at the end of the storage period of 150 days, respectively (DUYAR and EKE, 2009).

TMA values of the anchovy and sardine marinades were reported as 0.50 and 0.80 mg/100 g at the end of the storage period of 170 days by DUYAR and EKE (2009), respectively. These values are quite lower than our findings. It was thought that this situation was resulted from high TMA contents of the anchovies which were our raw material.

TBAR values of the fish ball marinated after frying are shown in Fig. 2. TBAR value of the fresh material was determined as 1.98 mg MA/ kg. The initial TBAR value of the fish ball increased from 1.71 to 5.68 MA/kg at the end of the storage period of 150 days. It was reported that, TBAR value that is used to determine rancidity in lipid is less than 3 mg MA/kg in a very good material and it must not be bigger than 5 mg MA/kg in a good material. Rancidity began if it exceeded 4 mg MA/kg and acceptability limit value is 7-8 mg MA/kG (CURRAN, 1980; VARLIK et al, 1993). Therefore it may be said that, the rancidity in the fish ball began on day 135. Similar to our study, DUYAR and EKE (2009) stated that the anchovy marinade which has 10.93 MA/kg of TBAR value at day 100 exceeded the acceptability limit values at day 130. There haven't been any studies investigating TBAR value of the fish ball marinated after frying. But there are studies related with marinades without frying (CADUN et al, 2005; DUYAR and EKE, 2009; BÍLGÍN et al., 2011; GUNSEN et al, 2011; SIMAT et al, 2011).

pH values of the fish ball marinated after frying are shown in Fig. 4.The pH value of the fresh material was determined as 6.1. The initial pH value of the fish ball that was 4.53 decreased to 3.42 at the end of the storage period of 150 days.

Microbiological quality of the fresh material and the fish ball marinated after frying is shown in Fig. 5, 6 and 7. The total mesophilic aerobic bacteria count of the fresh material that was 4.48 log cfu/g increased to 5.41 log cfu/g for the fish ball marinated after frying at the end of the storage period of 150 days. Total yeastmold count decreased along with the marination of the fresh anchovy; however it increased progressively throughout the storage.

Generally, Pseudomonas, Achromobacter and FLavobacterias are existing in newly caught fishes. Most of them have proteolitic properties. These microorganisms are sensitive to acid, so they are not existing in the marinades. Because development of these microorganisms is inhibited at about pH of 4.5. Betabacterium buchneri group of microorganisms are existing dominantly in the spoiled marinades. Beta-bacterias are isolated not only from cold marinades, but also from fried marinades and some special products with the cold marinade.

Because free aminoacids are in small amount, this situation does not occur too often in the fried marinades. The beta-bacterias and gamma-aminobutyric acid that is one of the reduced products in decarboxylation of aminoacids bind acetic acid and increase the pH value. The molds play important role only in the marinades kept in open cases (MEYER, 1965; KILINC and CAKLI, 2004). The total mesophilic bacteria of the fresh anchovy was determined as 4.8 log cfu/g by GUNSEN et al. (2011). There are not any studies investigating the microbiological quality of the fish ball marinated after frying. However several studies of the marinades without frying are existing (OLGUNOGLU, 2007; KURTKAYA, 2009; OZOGUL et al, 2009; PELIN CAN and ARSLAN, 2011).

Changes in the sensory quality and overall acceptability of the fish ball marinated after frying are shown in Table 1. Considering overall acceptability results of sensory analysis of the fish ball, it was seen that the fish ball having score of 9.6 indicated acceptable quality on day 135 of the storage period. And then the fish ball having score of 4.6 was evaluated as spoiled. Even though chemical and microbiological quality criteria did not exceed the acceptability limit values at the end of the storage periods of 150 days, the product spoiled in terms of sensory properties. Flavor and aroma development in the marinade is resulted from the degradation of protein and lipid with mutual effects of the acetic acid and salt with enzymes. (McLAY, 2001).

The shelf life of anchovy ball marinated in solution containing 6% salt and 1% vinegar after frying was determined as 120 days by VARLIK et al (2000). This result was similar to our study. In addition, there are several studies investigating the shelf life of the marinades without frying (AKSU et al. 1997; DOKUZLU, 1997; OZDEN and BAYGAR, 2003; DUYAR and EKE, 2009).

CONCLUSIONS

The shelf life of the fish ball marinated after frying was determined as 135 days. Even though the acceptability limit values were not exceeded according to chemical and microbiological analysis results, the product spoiled in terms of sensory properties. As a consequence, it might be suggested that the fish ball must not be consumed if it is stored over 4 months.

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Paper received June 17, 2013 Accepted November 4, 2013