QUALITY OF FISH SAUCE PRODUCTS FROM RECYCLED BY-PRODUCTS FROM FISH GEL AND KAMABOKO PROCESSING

QUALITY OF FISH SAUCE PRODUCTS FROM RECYCLEDBY-PRODUCTS FROM FISH GEL AND KAMABOKO PROCESSINGjfq_434 1..11

T. TAKANO1, K. SHOZEN2*, M. SATOMI3, W. TAIRA4†, H. ABE4‡ and Y. FUNATSU5,6

1Umekama Co., Ltd., Toyama, Japan2Toyama Food Research Institute, Toyama Prefectural Agricultural, Forestry & Fisheries Research Center, Toyama, Japan3National Research Institute of Fisheries Science, Yokohama, Japan4Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, Japan5Department of Food Science and Human Wellness, Rakuno Gakuen University, Ebetsu 069-8501, Japan

6Corresponding author. TEL:+81-11-388-4892; FAX: +81-11-387-5848;EMAIL: [email protected]

Present/permanent address: *Fisheries andFishing Port Division of Agriculture, Forestryand Fisheries Department, Toyama PrefecturalGovernment, Toyama 930-8501, Japan†Department of Primary Education, TsuruUniversity, Yamanashi 402-8555, Japan‡Research Institute of Seafood Biochemistry,Tokyo 167-0023, Japan

Received for Publication February 7, 2011Accepted for Publication December 24, 2011

doi:10.1111/j.1745-4557.2012.00434.x

ABSTRACT

In order to make effective reuse of kamaboko processing wastes and to minimize theamount of waste disposal, the production of two kinds of fish sauces was tried on asmall industrial scale from the wastes with or without the addition of the meat ofdeepsea smelt (Glossanodon semifasciatus). They were fermented for 6 months atroom temperature using salt and koji mold. As a control, a fish sauce was also pro-duced only from the deepsea smelt meat. The recovery of fish sauce from the initialmashed mixture ranged between 75 and 79%, depending on the ingredients in thethree fish sauce products. The total nitrogen content of the waste sauce and themixed sauce was lower than that of the control. The levels of the original additivesto kamaboko products, b-carotene and sorbic acid, were very low in these fish sauces.The taste-active components of the waste sauce and mixed sauces were lowerthan those of the control. Sensory evaluation revealed that the former two wastesauces were less bitter and higher in saltiness than the control. However, no differ-ence was found in umami taste between these products. These findings suggest thatthe wastes from kamaboko processing factories could be reused as fish sauce for foodcondiments.

PRACTICAL APPLICATIONS

Recent development in the food industry in Japan has enabled the surimi-basedproducts to be mass produced and standardized even in the fish gel, kamaboko trade.However, a serious problem has arisen in the development of kamaboko processingthrough the discharge of the wastes, i.e., nonstandard products or fragments, fromkamaboko processing factories. It is clear that the waste from kamaboko processingfactories is transformed effectively into fish sauce by using soy sauce koji mold. As aresult, the amount of the discharged wastes from kamaboko factories are able to beminimized because the liquefaction ratio of the fish sauce mushes (moromi) fromthe wastes after fermentation was high and the products have a high umami taste andagreeable soy sauce-like flavor.

INTRODUCTION

A variety of marine products have been consumed worldwidefor a long time. Particularly in Japan, the diversified eatinghabits have brought various kinds of marine products intothe market. The products are roughly categorized into six

product groups such as surimi-based products, frozenproducts, dried products, canned products, salted productsand fermented products (Funatsu 1998). Of these products,surimi-based products are widely and largely consumed inJapan and many kinds of the products such as fried, broiled,steamed, boiled and flavored kamaboko, and broiled chikuwa

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Journal of Food Quality ISSN 1745-4557

1Journal of Food Quality •• (2012) ••–•• © 2012 Wiley Periodicals, Inc.

have been produced for a long time (Park 2004). In this rangeof kamaboko products, maki kamaboko such as akamaki orkobumaki, which have rich tastes and soft textures, are kindsof steamed kamaboko and are the special products of theToyama Prefecture in Japan.

On the other hand, recent developments in the foodindustry have enabled the surimi-based products to be massproduced and standardized even in the kamaboko trade.However, a serious problem has arisen in the development ofkamaboko processing with the discharge of the wastes, i.e.,nonstandard products or fragments, from kamaboko process-ing factories. A prompt solution is required because theprice of raw material, frozen surimi, has been rising andcausing financial strain to factories. Annual disposal ofthe wastes even in a factory, Umekama Co., Ltd., in Toyama,amounted to 16.7 tons in 2006, although the seasonal changewas rather large. Most of the nonstandard products and frag-ments from kamaboko processing factories are used as live-stock, feeds and organic fertilizers, and the remainder isdisposed as industrial waste.

In the meat processing factories, however, the wastesfrom raw meat during processing of meat products such asham, sausage and bacon have been reused as raw materials forother meat products such as meatball and steak using adhe-sive technology with enzymatic reactions. However, wasterecyclability in surimi-based products is rather difficultowing to problems such as differences in ingredients, varyingsizes of the nonstandard products or fragments and bacterialproliferation in the waste during storage.

Fish sauces are the most important fermented fish prod-ucts used as traditional condiments in Southeast Asian coun-tries. However, these traditional fish sauces contain largeamounts of salt (>20%) and have a peculiar smell, althoughthe products have strong umami taste which is derived fromfree amino acids and oligopeptides originating from fish pro-teins (Funatsu et al. 2004a). There have been some attempts(Sakai 1993; Dohmoto et al. 2001; Osako et al. 2005) toimprove the taste and smell of fish sauces by using koji moldduring fermentation. Sakai (1993) tried to prepare condi-ments from fisheries factory waste to develop efficient usageof marine resources, i.e., trash meat from masu salmon andpink salmon, for the purpose of efficient usage of marineresources. Dohmoto et al. (2001) prepared fish sauces fromtwo kinds of fishes and a squid employing not only koji moldbut also lactobacilli and yeast for the multipurpose usageof fish sauces. Osako et al. (2005) reported that the taste offish sauces prepared from the raw meat of Japanese anchovyand rabbit fish using wheat malt were species-specific, but thetaste of those prepared from heated meat were similar to eachother. Uchida et al. (2005) reported that fish sauces preparedfrom Chinese silver carp with both koji mold and lactic acidbacteria or only the former received high scores by sensoryevaluation when compared with the fish sauces prepared

without koji mold or Thailand and Chinese commercial fishsauces.

For more efficient utilization of several underutilizedfishes, the authors (Funatsu et al. 2002, 2004a; Uchida et al.2005; Taira et al. 2007) have tried to produce fish sauces fromseveral fish species on a small industrial scale using soy saucekoji mold and have examined the characteristics of the prod-ucts from many angles. However, no information is availableon the characteristics of the fish sauces fermented using thewastes from kamaboko processing factories. In this study, wetried to produce fish sauces from the wastes from kamabokoand characterize the qualities of the products from someaspects such as physicochemical and microbial parameters,taste-active components, adenosine-5′-triphosphate (ATP)and its related compounds, volatile compounds and sensoryattributes. The qualities of these fish sauces were comparedwith those of fish sauce prepared from fish meat only.

MATERIALS AND METHODS

Materials

The wastes from kamaboko processing were collected andsubsequently stored frozen at -20C until use. The deepseasmelt (Glossanodon semifasciatus) was purchased from fishmarkets in Toyama and Niigata, Japan, and also stored frozenat -20C until use.

Preparation of Fish Sauce

After thawing, the wastes and whole fish were minced in ameat grinder. Minced wastes (260 kg) were mixed with 40 kgof soy sauce koji prepared by incubating 1:1 mixture ofsteamed defatted soybean and roasted wheat with Aspergillusoryzae (Ichimurasaki, Bio’c, Toyohashi, Japan), 62 kg of saltand 86 kg of water in a polypropylene tank of 500 L volumewith a cover (Table 1). The fish sauce prepared from the abovemixture was hereafter called “waste sauce.” Minced wastes(120 kg) and deepsea smelt meat (150 kg) were also mixedwith 40 kg of koji, 64 kg of salt and 86 kg of water in the sametype of tank as described earlier (Table 1). This fish sauceprepared from this mixture was called “mixed sauce.” As a

TABLE 1. INGREDIENT COMPOSITION OF FISH SAUCE PRODUCTS:DEEPSEA SMELT SAUCE (CONTROL), KAMABOKO WASTE SAUCE ANDTHE MIXTURE (MIXED SAUCE)

Deepseasmeltmeat (kg)

Kamabokowastes (kg)

Kojimold (kg)

Salt(kg)

Water(kg)

Control 260 0 40 66 86Waste sauce 0 260 40 62 86Mixed sauce 150 120 40 64 86

QUALITY OF FISH SAUCES FROM KAMABOKO PROCESSING WASTES T. TAKANO ET AL.

2 Journal of Food Quality •• (2012) ••–•• © 2012 Wiley Periodicals, Inc.

control, minced fish (260 kg) was mixed with 40 kg of the koji,66 kg of salt and 86 kg of water (Table 1). The salt concentra-tions between kamaboko and deepsea smelt meat weredifferent. The salt concentration of the kamaboko is about1.5%, although that of the deepsea smelt meat was very low.Therefore, final salt concentrations of the three fish sauceproducts before fermentation were adjusted almost at thesame level (14.3–14.7%, w/w). After mixing, these fish saucemushes (moromi) were subsequently fermented for 6 monthsat room temperature from April to September in a prefabri-cated house without air conditioner. The monthly average ofthe outer temperature from May to September in Toyama citywas about 7, 12, 17, 22, 25 and 26C, respectively. To achieveuniform fermentation, tanks were well stirred by a paddle atabout 2-week intervals. After fermentation, the three fishsauce mushes were put into nylon bags and squeezed by acompressor (KS-4, Shinkomagata Machinery, Tokyo, Japan).The fish sauce squeezed out was then heated up to 90C ina steam caldron (Hattori Industry, Okazaki, Japan). Aftercooling to room temperature, the fish sauce was filtered by acirculating filtration machine with membranes of 1.68 m2

area and 0.2 mm pore size (7/M-1, Noritake, Nagoya, Japan).In this study, part of the filtrate was bottled in plastic bottles asan experimental sample.

Analytical Methods

Color characteristics were determined by measuring L*, a*and b* values using a color difference meter (TC-8600, TokyoDenshoku, Tokyo, Japan). Hue angle (h) and saturation (C*)values were calculated with using a* and b* values. Totalnitrogen was determined by the Kjeldahl method. pH and saltcontents were determined according to the soy sauce analysismethod (Funatsu et al. 2004b). Trimethylamine nitrogen wasdetermined by the method of Yamagata et al. (1968). Sorbicacid levels were determined by AOAC Official Methods ofAnalysis (AOAC 1984), with a slight modification. Gas chro-matography (GC) analysis for the determination of the sorbicacid was carried out under the following conditions: column,Thermon 1000 (Chromosorb 80–100 mesh, length, 2.1 minner diameter [i.d.], 3.2 mm); column temperature, 160C;injection port temperature, 170C; GC detector, flame ioniza-tion detector; carrier gas, N2 (0.2 mL/min). b-Carotene levelwas determined by the method of Granelli and Helmersson(1996). Each specimen was diluted with deionized water, fil-tered through a cellulose membrane (0.45 mm) and used foranalysis as follows. Free amino acids were analyzed with anamino acid autoanalyzer (L-8500A, Hitachi, Tokyo, Japan).Organic acids were determined by high-performance liquidchromatography (Shimadzu, Kyoto, Japan). Six kinds oforganic acids were separated using a Shim-pack SCR-102Hcolumn (8 mm i.d. ¥ 300 mm, Shimadzu), eluted by 5 mMp-toluenesulfonic acid solution at 40C at a flow rate of

0.8 mL/min, and monitored at 210 nm. Histamine was deter-mined by using a Check Color Histamine Kit (Kikkoman,Noda, Japan) with enzymatic reactions (Sato et al. 2005).

Microbial Analyses

Fish sauce (10 g) was transferred aseptically from each finalproduct and homogenized in 90 mL of sterilized saline solu-tion containing 10% NaCl (w/v). Serial dilutions of homoge-nates were made and nonhalophilic bacterial counts weredetermined using standard count agar as the medium (EikenChemical, Tokyo, Japan). Halophilic bacteria were countedusing marine agar (Becton Dickinson, Franklin Lakes, NJ)containing 2% NaCl. Growth of Escherichia coli and othercoliform bacteria was determined on desoxycholate agar(Eiken Chemical). For heterotrophic plate counts and othercoliform bacteria, yeasts and molds, the incubation was at37C for 48 h. Microbial data are presented as colony-formingunits (cfu).

Volatile Compound Analyses

Five milliliters of sample of each specimen and 4.5 mL of 1%cyclohexanol (Wako Pure Chemical Industries, Osaka, Japan)were put into a glass vial (1.5 cm i.d. ¥ 5.0 cm length; Sigma-Aldrich, St. Louise, MO). After sealing, the vial was vibratedwith a test tube mixer for 30 s. A solid phase micro extrac-tion (SPME) fiber (75 mm carboxen/polydimethylsiloxane,Supelco, Tokyo, Japan) was inserted into the vial and the vola-tile compounds were absorbed on the fiber at 40C for 60 min.After absorption, the SPME fiber was injected into a gas chro-matograph. Gas chromatography/mass spectroscopy (GC/MS) analysis was carried out under the following conditions:gas chromatograph, Hewlett–Packard 6890 type (Hewlett–Packard, Palo Alto, CA); mass spectrometry detector,Hewlett–Packard PTA-5 column (0.32 mm i.d. ¥ 30 m; filmthickness, 1.5 mm); column temperature, 40C (2 min)-250C(23 min); carrier gas, He; injection port temperature, 250C;injection method, split less (flow rate, 1.5 mL/min); heatingrate, 10C/min; ion-trap manifold temperature, 230C; elec-tron impact ionization voltage, 70 eV. Peak concentrationvalues represent the ratios of the peak concentration in a fishsauce product to that of the internal standard (cyclohexanol).In the GC/MS analysis, the obtained peaks were identifiedusing the standard National Institute of Standard and Tech-nology mass spectrum database (Van den Dool and Kraft1963) and/or by comparing their retention times with thoseof the authentic compounds.

Sensory Evaluation

Sensory evaluation was conducted by trained Japanese panel-ists comprising 10 persons aged 20–30 years. Each assessment

T. TAKANO ET AL. QUALITY OF FISH SAUCES FROM KAMABOKO PROCESSING WASTES


was conducted according to the paired comparison test(Basker 2010). A set of test solutions was presented to eachpanel member in brown glass cups at room temperature. Thepanelists were requested to make an evaluation of the follow-ing items based on their taste and smell: sweetness, saltiness,sourness, bitterness, umami, first taste and aftertaste. Thesmell of fish sauce products was tested by sniffing (Fukamiet al. 2004). The panelists were asked to score the smell andtaste of each fish sauce from -3 to +3 in comparison with thatof a fish sauce prepared only from the deepsea smelt meat(control).

Statistical Analyses

Data for physicochemical and microbial parameters, ATP andits related compounds, and volatile compounds were statisti-cally analyzed using Dunnett’s test (jmp 8.0.2, SAS Institute,Tokyo, Japan) and the data for sensory evaluation were alsostatistically analyzed using the Mann–Whitney U-test.

RESULTS AND DISCUSSION

Physicochemical and Microbial Parametersof Fish Sauce Products

The recovery of fish sauce from the initial mush rangedbetween 75 and 79%, depending on the ingredients of threefish sauce products (Fig. 1). It is considered that the recoverybetweentheproductspreparedwithmanykindsof ingredientscould be similar in this study.However, there were some differ-ences in recovery between the present trial and our previous

one (Taira et al. 2007) due to the difference in fermentationconditions, fish species and the ingredients of the products.The recovery of fish sauce from deepsea smelt was lower thanthat of fish sauces from flying fish and common dolphinfish ofsmall size because the lipid content of the deepsea smelt,whichare caught in spring season, was high. However, the recoverywas slightly higher in the former two than the waste and mixedsauces in this study (Taira et al. 2007).

The lightness (L*) value of the waste and mixed sauceswas significantly higher (P < 0.05) than that of the control,although the hue angle (h) and saturation (C*) values ofthe former two were significantly lower (P < 0.05) than thoseof the latter, suggesting that the color of the products waspaler than the control with light yellowish brown (Table 2).The colors of nampla, nuocmam, patis and yeesui were dullorange, grayish orange and grayish bright yellow, respectively(Funatsu et al. 2004b). It is considered that these color differ-ences between fish sauces have a close relation to the Maillardreaction products during fermentation (Hondo 1993; Murataet al. 1995).

The pH values of the products were in the range from 4.6 to4.8, and were lower than those reported for fish sauces fromSoutheast Asian countries (Park et al. 2001) and Japanese-made fish sauces prepared traditionally without using kojimold (Funatsu et al. 2000). This may be due to the differencein the fermentation mechanisms between the samples withand without koji mold.

Control Waste sauce Mixed sauce0

10

20

30

40

50

60

70

80

90

100

Rec

over

y (%

)

FIG. 1. RECOVERY OF FISH SAUCE FROM THE INITIAL MUSH

TABLE 2. PHYSICOCHEMICAL AND MICROBIAL PARAMETERS OFTHREE FISH SAUCE PRODUCTS

ControlWastesauce

Mixedsauce

L* 78.00 86.75* 87.01*h 85.89 92.29* 92.61*C* 61.56 48.40* 47.27*pH 4.8 4.6 4.7Salt (%) 16.5 18.2 18.8Total nitrogen (g/100 mL) 2.0 1.5* 1.8Trimethylamine nitrogen (mg/100 mL) 10 3* 6*Sorbic acid (g/L) nd nd 0.03*Histamine (ppm) 108 88 237*b-Carotene (mg/100 mL) 1.1 2.1 1.8Nonhalophilic bacterial count (cfu/g) <30 <30 <30Coliform bacteria (cfu/g) <30 <30 <30Halophilic bacteria (cfu/g) <30 <30 <30Yeasts and molds (cfu/g) <30 <30 <30

Color characteristics were measured with a 4-mm glass cell.See Table 1 for a detailed description of the control, the waste sauce andthe mixed sauce.Values are means from three different experiments. The significancebetween the control, waste sauce and mixed sauce was analyzed Dun-nett’s test: *P < 0.05.nd, below detection limit (<0.01 g/L); h = tan-1(b*/a*); C* = [(a*)2 +(b*)2]1/2.



The salt concentration of the waste and mixed sauces wasslightly higher than that of the control. This may be due to thedifference in salt contents between deepsea smelt meat andkamaboko wastes and the difference in lipid contents ofthe underutilized fishes that have various sizes and fishingarea. Total nitrogen contents of the fish sauces produced inthis study fall into grade II according to the Thai fish saucestandard (Brillantes and Samosorn 2001) and also fall intohigh grade according to the Japanese Agricultural Standardof Soy Sauce (Japanese Agricultural Standard of Soy Sauce2009). Trimethylamine nitrogen level was significantly lower(P < 0.05) in the waste and mixed sauces than in the control.In general, the main materials of kamaboko are frozen surimi.The important step of surimi processing to ensure odorlessand colorless surimi, as well as gel forming ability, is efficientwashing (Park and Morrissey 2001). Some differences intrimethylamine nitrogen levels between control and othersamples might be related to the mixing ratio of frozen surimiand deepsea smelt meat.

Sorbic acid and b-carotene levels of the fish sauce productswere below 0.03 g/kg and 1.1–1.9 mg/100 mL, respectively.The small amounts of sorbic acid and b-carotene werederived from the additives of original kamaboko as a preserva-tive and colorant, respectively.

Histamine contents of the products ranged from 88 to237 ppm. According to Funatsu et al. (2004a) and Brillantesand Samosorn (2001), the histamine contents of the commer-cial fish sauces varied from 74 to 851 ppm. Thus, the hista-mine levels in the present products fall in this range. However,a high level of histamine accumulation (>1,000 ppm) wasfound in one of the fish sauce mushes prepared from smalldolphinfish (Coryphaena hippurus) during fermentation asdescribed in our previous report (Taira et al. 2007). One ofthe major histamine-producing bacteria during fish saucefermentation is thought to belong to the genus Tetragenococ-cus (Ito et al. 1985; Satomi et al. 1997; Kimura et al. 2001).Satomi et al. (2008) reported that a predominant histamine-producing bacteria during fish sauce fermentation wasidentified to be a strain H of Tetragenococcus halophilus byphenotypic and 16S rRNA analyses. Therefore, it is possiblethat T. halophilus H strain is closely related to histamine accu-mulation during our previous fish sauce fermentation (Tairaet al. 2007). Shozen et al. (2010) reported recently that theaddition of T. halophilus to the mush of the wastes fromkamaboko processing and koji mold at the beginning of fishsauce fermentation promoted a rapid decline of pH at anearly stage of fermentation and restrained the proliferation ofhistamine-producing bacteria. As a result, the histamine levelin the fish sauce mush was below 50 ppm throughout the fer-mentation. These facts indicate that some of the ingredientsof the wastes could be responsible for the control of histamineaccumulation during fish sauce fermentation. Nonhalophilic,halophilic and coliform bacteria as well as yeasts and molds

were under the detection limit (<30 cfu/g) in all fish sauceproducts.

Taste-Active Components of FishSauce Products

The free amino acid and organic acid compositions of thethree fish sauce products are shown in Tables 3 and 4, respec-tively. The total free amino acid levels of the waste and mixedsauces (5,753 and 7,197 mg/100 mL, respectively) were lower

TABLE 3. FREE AMINO ACID COMPOSITIONS OF THREE FISH SAUCEPRODUCTS

(mg/100 mL, mean of duplicate analysis)

ControlWastesauce

Mixedsauce

Taurine 144 18 57Aspartic acid 199 123 191Threonine 565 230 354Serine 575 267 345Glutamic acid 1,345 1,299 1,219Glycine 292 380 300Proline 360 153 220Alanine 1,366 550 830Valine 757 297 460Cystine 25 nd ndMethionine 387 180 229Isoleucine 723 305 449Leucine 1,149 584 784Tyrosine 93 77 88Phenylalanine 586 248 348Tryptophan 14 nd 11Ornithine 603 222 347Lysine 1,192 617 776Histidine 230 103 105Arginine 83 100 86Total 10,687 5,753 7,197

See Table 1 for a detailed description of the control, waste sauce andmixed sauce.nd, not detected.

TABLE 4. ORGANIC ACID COMPOSITION OF FISH SAUCE PRODUCTS

(mg/100 mL, mean of duplicate analysis)

Control Waste sauce Mixed sauce

Malic acid 32 34 7Succinic acid 7 81 6Lactic acid 1,598 1,218 1,345Fumaric acid 7 2 5Acetic acid 92 78 180Pyroglutamic acid 526 388 226Total 2,262 1,801 1,768

See Table 1 for a detailed description of the control, waste sauce and themixed sauce.



than those of the control (10,687 mg/100 mL). Taurine levelsof the waste and mixed sauces were also far lower than thoseof the control. Essential amino acid levels in the waste andmixed sauces (summed up to 2,164 and 2,991 mg/100 mL,respectively) were lower than in the control (4,787 mg/100 mL), although tryptophan was similar in both mixedsauce and control sample. The reason why the waste andmixed sauces showed lower amino acid values compared withcontrol may originate from the difference in protein contentsbetween fish meat having high protein and kamaboko wasteshaving low protein contents.

Total organic acid contents of the waste and mixed sauceswere also lower than those of the control. Lactic acid, thehighest organic acid component of the products, was lower inthewasteandmixedsauces(1,218and1,345 mg/100 mL)thanin the control (1,598 mg/100 mL).The larger amount of lacticacid in the control was due to the minced meat, which origi-nally contained a large quantity of lactic acid, and the produc-tion of lactic acid by lactic acid fermentation. Other organicacids, however, were not always high in the control fish sauce.For instance, malic and acetic acid are the highest in the wasteand the mixed sauce, respectively, suggesting the differentmicrobial activities in each fish sauce during fermentation.The pyroglutamic acid in the waste and mixed sauces wasfound at high concentrations compared with the control.

Park et al. (2002) reported that 11 compounds, whichconsist of glutamic acid, aspartic acid, threonine, alanine,valine, histidine, proline, tyrosine, cystine, methionine andpyroglutamic acid, were identified to be the taste-active com-ponents in a Vietnamese fish sauce, and these compoundsmight contribute to umami, sweetness and overall taste of thefish sauce. Figure 2 shows the star diagram of the contentsof taste-active components of three fish sauce products.Control, wastes and mixed sauces show somewhat differentswallow-like patterns due to the different contents of eachcomponent between these three products. Although the

alanine and pyroglutamic acid levels were different, the wastesauce and the mixed sauce showed a similar pattern.Accordingto the data from Tables 3 and 4, the total amounts of the taste-active components in the control,waste and mixed sauces were5,853, 3,400 and 3,922 mg/100 mL, respectively. Therefore,the taste strength of the waste and mixed sauces might be dif-ferent from that of the control. Although the taste strength ofthe former two is estimated to be low, this disadvantage wouldbe improved by mixing the kamaboko wastes with high ratio ofdeepsea smelt meat containing high protein content.

The level of ATP and its related compounds of threefish sauce products are shown in Table 5. Although noneof ATP, adenosine-5′-diphosphate ADP, adenosine-5′-monophosphate (AMP), inosine-5′-monophosphate (IMP),

Glutamic acid

Aspartic acid

Threonine

Alanine

Valine

HistidineProline

Tyrosine

Cystine

Methionine

Pyroglutamic acid1500

500

1000

Glutamic acid

Aspartic acid

Threonine

Alanine

Valine

HistidineProline

Tyrosine

Cystine

Methionine


500

1000


Glutamic acid

Aspartic acid

Threonine

Alanine

Valine

HistidineProline

Tyrosine

Cystine

Methionine


500

1000

FIG. 2. STAR DIAGRAM OF TASTE-ACTIVE COMPONENTDS IN FISH SAUCE PRODUCTS, THE WASTE AND MIXED SAUCES, AND THE CONTROLData are compiled from Tables 4 and 5.

TABLE 5. THE LEVEL OF ATP AND ITS RELATED COMPOUNDS OF THREEFISH SAUCE PRODUCTS

(mg/100 mL)


ATP nd nd ndADP nd nd ndAMP nd nd ndGMP nd nd ndIMP nd nd ndHxR nd nd ndHx 58.9 45.5* 36.9*

* Waste sauce or mixed sauce groups whose means are significantlydifferent from the mean of the control group.See Table 1 for a detailed description of the control, waste sauce andmixed sauce.Values are means from three different experiments. Refer to the legend ofTable 2 for values and significance.ADP, adenosine-5′-diphosphate; AMP, adenosine-5′-monophosphate;ATP, adenosine-5′-triphosphate; GMP, guanosine-5′-monophosphate;Hx, hypoxanthine; HxR, inosine; IMP, inosine-5′-monophosphate; nd, notdetected.



guanosine-5′-monophosphate (GMP) and inosine (HxR)was detected in any of the specimens, hypoxanthine (Hx) wasdetected in all products. AMP, IMP and GMP, which areknown to contribute to umami taste, have not been detectedat all in fish sauces. Sado and Michihata (1996), Park et al.(2001), Osako et al. (2005) and Funatsu et al. (2004a) alsodetected none of AMP, GMP and IMP in all fish sauces exam-ined, i.e., Southeast Asian, Japanese traditional and Chinesefish sauces, and also Japanese new type fish sauces with kojimold. These facts suggest that there is no participation ofAMP, GMP and IMP in the umami taste of fish sauces, includ-ing those produced in the present experiment.

Volatile Compounds of Fish Sauce Products

The volatile compounds detected in fish sauce products areshown in Table 6. As a whole, 47 different volatile compoundswere identified by the SPME method and GC/MS analysis.According to some references (Mclver et al. 1982; Sancedaet al. 1984, 1986; Yokotsuka et al. 1998; Michihata et al. 2002),most of the volatile compounds detected in this study hadalready been detected by other methods such as steam distil-lation (Table 6). Thirty-five different volatile compoundsshown in Table 6 were also already identified by the SPMEmethod and GC/MS analysis (Funatsu et al. 2002). However,only five compounds, i.e., sorbic acid, n-nonanal, n-decanal,3-ethyl-phenol and ethyl caproate, were newly identified inthis study. Many kinds of volatile compounds, which werefound in soy sauce, were also detected in three fish sauce prod-ucts owing to the usage of koji mold in our preparations.

In acids, high level of isovaleric acid was commonlydetected in all samples. Isovaleric acid is one of the main acidsin fish sauce and is detected in fish sauce with and without kojimold (Funatsu et al. 2002). All samples of fish sauces hadisovaleraldehyde, 2-methylbutyraldehyde, 3-(methylthio)-propanal and benzaldehyde. Aldehydes have been derivedfrom lipid oxidation during fermentation, and branched,short-chain aldehydes or aromatic aldehydes might haveresulted from deamination of amino acids (Karahadian andLindsay 1989). These aldehydes are generally considered tocause unpleasant oxidation flavors in foods (Heath and Rei-neccius 1986).

In the waste and mixed sauces, many kinds of alcohols andesters were found. Particularly, the levels of alcohols such asethanol, isobutyl alcohol, isoamyl alcohol, 3-(methylthio)-1-propanol and phenylethyl alcoho; phenols such as guaiacol,3-ethyl-phenol and 4-ethyl-2-methoxy-phenol; esters such asethyl acetate and ethyl lactate; and lactones such as butyrolac-tone and g-valerolactone were significantly higher (P < 0.01)in the waste sauce than in the control. As the waste sauce con-tained 1.2% lactic acid, these alcohols might be obtained bymicrobial fermentation such as that of lactic acid bacteria.Esters are found in most fermented products and might be

products of esterification of alcohols with carboxylic acidsformed by microbial or enzymatic decomposition of lipid(Cha and Cadwallader 1995).

Nitrogen-containing compounds such as methylpyrazineand 2,5-dimethylpyrazine were detected only in the control. Itis well known that methylpyrazine and 2,5-dimethylpyrazineare related to nutty and/or roasty aroma (Qin et al. 2011).These compounds, which were only detected in the control,could be formed during fermentation and heating process.Therefore, volatile compounds of the waste and mixed saucesmight be considerably different from those of the controlowing to the differences in principal ingredient compositionsand food additives in the kamaboko wastes.

Trimethylamine, which was found in our previous report(Funatsu et al. 2002), could not be detected in this study.Although the broad peak that appeared in the range from 1 to2 min was considered to be trimethylamine, the peak wasnot exactly identical to that of authentic amines (datanot shown). From the results of trimethylamine nitrogencontents of the products shown in Table 2, the waste andmixed sauces (3–6 mg/100 mL) contain significantly lower(P < 0.05) trimethylamine than the control (10 mg/100 mL).This level in the control was slightly higher than that in theprevious report (3.2–6.2 mg/100 mL; Osako et al. 2005). It isthought that this difference may be derived from the differ-ence in freshness of the fish meat used as raw materials.

Nonomura et al. (1976) and Yokotsuka et al. (1998)reported that some volatile compounds such as 4-hydroxy-2(or5)-ethyl-5(or2)-3(2H)-furanone (HEMF), 4-hydroxy-2,5-dimethyl-3(2H)-furanone and 4-hydroxy-5-methyl-3(2H)-furanone, which have a strong caramel flavor, wereunique compounds found in soy sauce. However, these com-pounds were not detected in the present products. Yoshikawaet al. (2010) also reported that inoculation with halotolerantbacteria as a starter and koji initially in chum salmon saucefermentation was effective to improve the color and umamitaste of the products and to confer soy sauce-like flavorsuch as 2-phenylethyl alcohol, 4-ethyl-2-methoxy-phenoland HEMF on the fish sauce. Although the former two weredetected in the present fish sauces products, the latter was notdetected in any of the products.

Sensory Evaluation of Fish Sauce Products

To confirm the taste and smell characteristics of theseproducts, sensory evaluation was conducted using the controlfish sauce as a reference (Table 7). There was a significantdifference (P < 0.01) in smell between the waste sauce andthe control. Trimethylamine nitrogen level was significantlylower (P < 0.05) in the waste and mixed sauces than in thecontrol (Table 2). Thus, it is estimated that fishy odor could beweaker in the waste and mixed sauces than in the control.According to SPME method and GC/MS analysis mentioned



TABLE 6. VOLATILE COMPOSITIONS OF THREE FISH SAUCE PRODUCTS

Compounds Control Waste sauce Mixed sauce References

AcidsAcetic 0.027 � 0.008 0 0.120 � 0.068** a, b, c, d, e, fIsovaleric 0.244 � 0.004 0.169 � 0.021** 0.137 � 0.019** a, e, f2-Methyl-butanoic 0 0.110 � 0.025** 0 e, fIsocaproic 0.002 � 0.001 0** 0** a, e, fSorbic 0 0 0.046 � 0.016** NPRn-Decanoic 0.009 � 0.001 0.040 � 0.008* 0.020 � 0.010 b

AldehydesIsovaleraldehyde 0.041 � 0.021 0.054 � 0.005 0.081 � 0.019** e, f2-Methylbutyraldehyde 0.084 � 0.027 0.170 � 0.006** 0.222 � 0.017** d, e, f3-(Methylthio)-propanal 0.030 � 0.005 0.072 � 0.022 0.046 � 0.035 d, eBenzaldehyde 0.011 � 0.003 0.046 � 0.015** 0.025 � 0.010 d, e, fBenzeneacetaldehyde 0.006 � 0.002 0.017 � 0.006** 0 e, fn-Nonanal 0.004 � 0.003 0.013 � 0.005** 0.007 � 0.002 NPRn-Decanal 0 0.005 � 0.002** 0.002 � 0.001 NPR(2Z)-2-Phenyl-2-butenal 0 0.005 � 0.001** 0 f

Sulfur-containing compoundsDimethyl trisulfide 0 0 0.006 � 0.002** d, f

AlcoholsEthanol 0.012 � 0.005 0.671 � 0.440** 0.201 � 0.042 a, b, c, d, e, fIsobutyl alcohol 0 0.182 � 0.002** 0 e, f1-Penten-3-ol 0.037 � 0.005 0** 0.084 � 0.004** d, e, f3-Pentanol 0.055 � 0.007 0** 0** fIsoamyl alcohol 0.127 � 0.010 1.210 � 0.066** 0.092 � 0.009 a, b, d, e, f2-Methyl-1-butanol 0 0.014 � 0.002** 0 e, f2,3-Butanediol 0 0.406 � 0.129** 0 a, e, f1-Pentanol 0.006 � 0.001 0** 0** a, d, e, fFurfuryl alcohol 0.335 � 0.005 0.241 � 0.050 0.240 � 0.048** a, b, d, e, f3-(Methylthio)-1-propanol 0.010 � 0.001 0.335 � 0.101** 0.010 � 0.004 e, fEthylhexanol 0.011 � 0.006 0.025 � 0.008** 0.006 � 0.001 d, eBenzyl alcohol 0.002 � 0.001 0 0.004 � 0.001 b, c, e, fPhenylethyl alcohol 0.012 � 0.001 0.391 � 0.117** 0.012 � 0.005 c, e, f

Nitrogen-containing compoundsMethylpyrazine 0.010 � 0.003 0** 0** d, e, f2,5-dimethylpyrazine 0.026 � 0.001 0** 0** a, b, c, d, e, f

Ketones2-Butanone 0.004 � 0.002 0** 0** b, d, e, f3-Hydroxy-2-butanone 0 0.018 � 0.001** 0 a, d, e, f2-Hydroxy-1-phenyl-ethanone 0 0 0.004 � 0.003** eMethyl pyrrol-2-yl ketone 0 0.023 � 0.007** 0 e

PhenolsGuaiacol 0.001 � 0.0001 0.005 � 0.002** 0.004 � 0.001 f3-Ethyl-phenol 0.0001 � 0.0003 0.002 � 0.001** 0.001 � 0.001 NPR4-Ethyl-2-methoxy-phenol 0.005 � 0.0003 0.009 � 0.003** 0.002 � 0.001 c, f

EstersEthyl acetate 0.025 � 0.017 0.078 � 0.001** 0.076 � 0.010** b, c, d, e, fEthyl lactate 0.071 � 0.003 0.621 � 0.074** 0.120 � 0.030 c, d, e, f2-Ethoxyethyl acetate 0 0.007 � 0.003 0 fMethyl Benzoate 0.001 � 0.0001 0** 0** eDiethyl succinate 0 0.005 � 0.002 0 fEthyl caproate 0.003 � 0.001 0.032 � 0.010 0.011 � 0.002** NPR

Furans2-Ethylfuran 0.062 � 0.013 0.008 � 0.002** 0.092 � 0.012** e, f

LactonesButyrolactone 0 0.040 � 0.010** 0.008 � 0.007 fg-Valerolactone 0.003 � 0.0001 0.109 � 0.024** 0.007 � 0.005 fg-Ethylbutyrolactone 0.010 � 0.001 0** 0** f

Values are means � standard deviation from three different experiments. The significance between the control, waste sauce and mixed sauce was ana-lyzed using Dunnett’s test: *P < 0.05; **P < 0.01.References: a, Mclver et al. (1982); b, Sanceda et al. (1984); c, Sanceda et al. (1986); d, Michihata et al. (2002); e, Funatsu et al. (2002); f; Yokotsukaet al. (1998).See Table 1 for a detailed description of the control, waste sauce and mixed sauce.NPR, not previously reported.



earlier, there were significant differences (P < 0.01) in volatilecompounds between waste sauce and the control (Table 5).Particularly, the contents of ethanol, isobutyl alcohol, isoamylalcohol, 3-(methylthio)-1-propanol, phenylethyl alcohol,4-ethyl-2-methoxy-phenol, ethyl acetate, ethyl lactate, g-butyrolactone and g-valerolactone were higher in the wastesauce than in the control. These volatile compounds hadalready been detected in soy sauce (Table 5). These findingssuggested that a smell similar to that of soy sauce (Yokotsukaet al. 1998; Funatsu et al. 2002) might be formed in the wastesauce during fermentation.

Compared with the taste of the control, the waste sauce wassignificantly lower in sweetness (P < 0.05) and bitterness(P < 0.01) and higher in saltiness (P < 0.05), while mixedsauce was significantly lower in bitterness (P < 0.01) andhigher in sourness (P < 0.01) and saltiness (P < 0.05). Therewas no significant difference (P > 0.05) in umami, first tasteand aftertaste between the products. Thus, it is consideredthat differences in the taste between the fish sauce productscould have originated from the different compositions of rawmaterials and from the activities of microorganisms duringfermentation.

CONCLUSIONS

It is clear that the waste from kamaboko processing factories istransformed effectively into fish sauce by using soy sauce kojimold in this study. As a result, we have found that it is possibleto minimize the amount of the discharged wastes from kam-aboko factories because the liquefaction ratio of the fish saucemushes (moromi) from the wastes after fermentation washigh (>75%) and the residues are possible to be reused forfeed and fertilizer (Funatsu et al. 2004a). The quality of theproducts was very high, retaining only low levels of food addi-tives derived from the original kamaboko such as sorbic acidand b-carotene and having a very low off-flavor or fishy odorbut high umami taste and agreeable soy sauce-like flavor.

ACKNOWLEDGMENTS

This research was supported by a fund from the JapaneseScience and Technology Agency for the Research for Pro-moting Technological Seeds (No.01-100) during fiscal year2006. The authors are grateful to Mr. T. Imai, former chiefof Toyama Prefectural Food Research Institute, Mr. K. Okui,president of Umekama Co., Ltd., and Dr. I. Kimura, formerdirector of the Fisheries Research Agency for their encour-agement and kind support. The authors thank Mr. D. A.Miller of Rakuno Gakuen University for proofreading thisarticle.

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QUALITY OF FISH SAUCE PRODUCTS FROM RECYCLED BY-PRODUCTS FROM FISH GEL AND KAMABOKO PROCESSING