Article

Development of Quality Index Method for anchovy(Engraulis anchoita) stored in ice: Assessment of itsshelf-life by chemical and sensory methods

AE Massa1,2, E Manca2 and MI Yeannes1,3

AbstractThe objective of this study was to develop a quality index method for Engraulis anchoita stored in ice and todetermine its shelf-life based on this quality index method and chemical indices such as total volatile basesand thiobarbituric acid-reactive substances. Besides, the chemical composition with emphasis on the poly-unsaturated fatty acids content was determined. The results indicate that E. anchoita is a valuable proteinsource and lipid with important content of n-3 polyunsaturated fatty acids. The developed quality indexmethod scheme was composed of 28 demerit points, divided into 4 parameters and 10 attributes. All attri-butes showed correlation with time of storage (R> 0.90). The quality index (QI) presented a linear relationshipwith storage (QI¼ 2.55x days in iceþ 1.76; R2

¼ 0.98). In the shelf-life assessment-based quality indexmethod, the rejection sensory point was observed after 7 days of storage due to the presence of unpleasantodours and deteriorated appearance. The total volatile basic nitrogen value remained below the upper limit ofacceptability during the 10 days of ice storage. The evolution of thiobarbituric acid-reactive substances indi-cates lipids oxidation during the storage of anchovies. According to the results, the quality index methodscheme developed for the E. anchoita stored in ice could be considered adequate to evaluate their freshnessand to estimate its shelf-life.

KeywordsEngraulis anchoita, quality index method scheme, ice storage, shelf-life

Date received: 7 March 2011; revised: 22 July 2011

INTRODUCTION

Engraulis anchoita is a small pelagic fish, which is foundin the South Western Atlantic Ocean (SWAO), fromVitoria (22 �S) in Brazilian waters to San Jorge Gulf(47 �S) in Argentine waters (Hansen et al., 2006;Pastous-Madureira et al., 2009). This pelagic specie ismore abundant in cold and low-salinity waters, but alsoinhabits in regions of higher temperatures and salinity(8–23 �C; 14–35 g/L) such as the waters facing thesouth-eastern basin of Brazil. This specie represents aviable commercial alternative for this region due itshigh biomass values and the decline of demersal fishery

(Hansen et al., 2006; Vas et al., 2007). In Argentina, thecommercial exploitation of E. anchoita is seasonal. Thehighest catch occurs during spring time in coastal sec-tors of Buenos Aires Province, where massive spawningoccurs (Pastous-Madureira et al., 2009). In the lastyears, catches were greatly increased, reaching 30,000tons (SAGPyA, 2009). This species is mainly exportedto European Union countries as ripened semipreserved

1Consejo Nacional de Investigaciones Cientıficas y Tecnicas(CONICET), Argentina2Instituto Nacional de Investigacion y Desarrollo Pesquero(INIDEP), Argentina3Departamento de Ingenierıa Quımica, Facultad de Ingenierıa,Universidad Nacional de Mar del Plata, Argentina

Corresponding author:AE Massa, Consejo Nacional de Investigaciones Cientıficas yTecnicas (CONICET), Argentina; Instituto Nacional deInvestigacion y Desarrollo Pesquero (INIDEP), ArgentinaEmail: aguedamassa@inidep.edu.ar

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anchovies by a salting and ripening process (Cabreret al., 2002; Yeannes and Casales, 2008). A smaller pro-portion of it is used by the canning industry and hasbeen started to be exported as marinated anchovies(Yeannes and Casales, 2008).

The chemical composition of fish varies between andwithin species depending upon several factors such asfood availability, the season, localisation, sex and age(Akpınar et al., 2009; Exler et al., 1975; Gorgun andAkpınar, 2007; Rueda et al., 1997; Shearer, 1994). Thisvariation will influence the quality of the fish productsand could be decisive in the yield and the application oftechnological processes (Huss, 1995; Stansby, 1967;Yeannes and Almandos, 2003). E. anchoita is a goodprotein source and lipids with high content of n-3 fattyacids, especially eicosapentaenoic acid (EPA, C20: 5n-3) and docosahexaenoic acid (DHA, C22: 6n-3)(Garcia-Torchelsen et al., 2009; Massa et al., 2009).These fatty acids play important roles in humanhealth and nutrition (Sahena et al., 2009; Simopoulos,2009; Valenzuela and Sanhueza, 2008). Great vari-ations were reported in the fat content and fatty acidscomposition for this anchovy. These variations wererelated to the life cycle of the fish and to external fac-tors like temperature, salinity and food availability(Chiodi, 1970; Garcia-Torchelsen et al., 2009; Massaet al., 2007, 2009).

Freshness is another important aspect to be con-sidered in the fishing industry. After capture, there isa progressive decline in the sensory and nutritionalquality of the fish due to intrinsic chemical and physicalchanges (called autolysis) and bacterial growth (Haard,1992, Huss, 1995; Kyrana and Lougovois, 2002; Ozogulet al., 2005; Surette et al., 1988). These processes gen-erate different organic compounds affecting the qualityof seafood (Haard, 1992; Howgate, 2010a; Huss, 1995;Ozogul et al., 2006; Pedrosa-Menabrito andRegenstein, 1988; Sykes et al, 2009). The rates and pat-terns of changes depend on several factors such as spe-cies, spawning, feeding habits, temperature and watersalinity, catching methods and storage conditions(Alasalvar et al., 2002; Ashie et al., 1996; Gill, 1990;Huss, 1995; Howgate, 2010a; Olafsdottir et al., 2004).Biochemical, microbiological and sensory methodshave been used to assess freshness and quality of fishduring handling and storage (Huss, 1995;Koutsoumanis et al., 2002). Sensory evaluation is themost satisfactory method to determine the freshnessand quality of fish and fish products, since it allowsassessment of autolytic and microbiological changes(Alsalvar et al., 2002; Barbosa and Vaz-Pires, 2004;Connell, 1995; Howgate, et al., 1992; Howgate,2010b; Huss, 1995; Hyldig and Green-Petersen, 2004).

The quality index method (QIM) has been intro-duced and widely studied as an alternative to others

sensory methods traditionally used (Botta, 1995;Bremner, 1985; Costell, 2002; Sveinsdottir et al., 2003;Sykes et al., 2009). The QIM is a fast and simplemethod to determine freshness in fishery products.Originally it was developed by researchers from theTasmanian Food Research Unit (Bremner, 1985).This method is based on the objective evaluation ofcertain sensory parameters of raw fish (skin, eyes,gills, etc.) that significantly changes during degradationprocesses (Cardenas Bonilla et al., 2007; Hyldig andGreen-Petersen, 2004; Olafsdottir et al., 1997;Sveinsdottir et al., 2003). The set of descriptors ofeach attribute is assigned a demerit points range (0–3), which are in direct proportion to their importancein the deterioration pattern of the species. The demeritscore awarded to each parameter are added together togive a total sensory score, called quality index (QI). Inthis way, a QI of zero is given for fresh fish and anincreasingly higher score as the fish deteriorates. Theobjective is to obtain a linear correlation between QIand storage time in ice, which makes it possible to pre-dict the remaining storage life (Cardenas Bonilla et al.,2007; Hyldig and Green-Petersen, 2004; Luten andMartinsdottir, 1997; Martinsdottir, et al., 2001;Sveinsdottir et al., 2003). Other important characteris-tic of QIM is that it is developed for each species andfishery product, because it takes into account their par-ticular characteristics. Therefore, it is necessary todevelop specific QIM scheme for each specie and fishproduct. In the same way, is needed evaluate the applic-ability of the QIM for fish stored under different con-ditions (frozen–thawed, ice slurry, superchilling,modified atmosphere packaging, etc.). A list of the spe-cies for which QIM schemes have been developed wascompiled by Barbosa and Vaz-Pires (2004), Hyldig andGreen-Petersen (2004) and Cyprian et al. (2008), amongother authors. QIM Eurofish team has developed amanual for the fish sector (Martinsdottir et al. 2001),available in 11 languages. This manual contains QIMschemes for 12 fish species and information about howto use the QIM schemes. Also, more than 50 scientificpublications are available on the website QIM Eurofish.So far, few QIM schemes have been developed withspecies of commercial interest of Latin-American: floun-der, Paralichthys patagonicus (Massa et al., 2005); whitecroaker (Micropogonias furnieri) in ice stored (Massaet al., 2006; Teixeira et al., 2009) and common carp,Cyprinus carpio (Agueria et al., 2007). So, the develop-ments of QIM scheme for other important fish speciesfrom Latin America are needed to monitor and studytheir quality.

One of the principal chemical changes that occur inmarine fish during spoilage is the production of volatileamines. These molecules are responsible for odour andflavour typical of fish with several days catch and they

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are commonly used as criteria for assessing the fishquality (Castillo-Yanez et al., 2007; El Marrackchiet al., 1990; Huss, 1995; Karungi, et al., 2004;Sikorski, 1990). The designation ‘volatile amines’regroups mostly three molecules, ammonia, dimethyla-mine (DMA) and trimethylamine (TMA). DMA andTMA result from the degradation of trimethylamineoxide (TMAO), a naturally occurring compound inmarine fish that plays an important role in the osmo-regulation process. DMA is mostly produced byendogenous enzymes while TMA is formed by activityof specific spoilage bacteria (Etienne, 2005; Huss,1995). DMA is present in low concentration in fishthat is freshly caught, it can be used to monitor fishfreshness and to evaluate the quality of frozen-storedproducts (Etienne, 2005; Howgate 2010c; Huss, 1995).TMA is an excellent spoilage index; is useful for object-ively measuring the eating quality of various fishesespecially on the medium-later phases of spoilage butit does not reflect the earlier stages of spoilage (Etienne,2005; Howgate 2010c; Huss, 1995). Total volatile basesnitrogen (TVB-N) was one of the first chemical indexesapplied to evaluate spoilage of fish (Dalgaard, 2000;Huss, 1995; Olafsdottir et al., 1997). This index includesthe most important nitrogenous volatile compoundsthat are present in fish under storage condition. TheTVB-N level in fish that is freshly caught is between 5and 20mg N/100 g muscle and levels of 30–35mg N/100 g muscle are considered the limit of acceptabilityfor ice-stored cold water fish (Connell, 1995;Dalgaard, 2000, Etienne, 2005; Howgate, 2010c;Huss, 1995). The European Union has established thelevel ranging from 25 to 30mg of TVB-N/100 g (EU,CEE/95/149).

During the ice storage of fish, significant changesalso occur in the lipid fraction. The marine lipids com-prise highly unsaturated fatty acids that are known tobe prone to oxidation (Chaouqy et al., 2008; Church,1998; Mazorra-Manzano et al., 2000; Xiong, 1994). Inthe advanced stages of oxidation, the breakdown ofhydroperoxides generates low molecular weight car-bonyl compounds and alcohol that lead to appearanceof objectionable odours and colours (Aubourg et al.,2005, 2007; Chaijan et al., 2006; Sikorski, 1990).Therefore, it is important to determine the fatty acidprofile to determine the degree of unsaturation and itssusceptibility to oxidative rancidity. Several methodshave been developed to assess lipid oxidation infoods. The thiobarbituric acid-reactive substances(TBARS) test is among the most widely used methodsused to quantify lipid oxidation products and it is asimple and fast test (Nishimoto et al., 1985; Ozogulet al., 2007; Papadopoulos et al., 2003; Tarladgiset al., 1960). The TBARS test determines the amountof malondialdehyde (MDA), a major secondary by-

product of lipid oxidation. The determination ofMDA in fish products has been widely studied, andsignificant correlations between the TBARS valuesand sensory assessment have been reported (Chaijanet al., 2006; Hoyland and Taylor, 1991; Undelandet al., 1999).

The aim of this study was to develop a QIM for E.anchoita during ice storage. Also, the freshness andremaining shelf-life for this specie during ice storagewas estimated based on QIM scheme developed andchemical indices.

MATERIALS AND METHODS

Samples were collected during research cruise con-ducted by the INIDEP on board the RV ‘Oca Balda’with a midwater trawl net. The study area included thecoastal sector of Buenos Aires province between 38 �Sand 41LS during the months of October–November,period of commercial catch.

Methods

Sample preparation. For chemical composition ana-lysis, the anchovies were placed in polyethylene bags;these were vacuum closed and frozen at �20 �C untilfurther analysis. The sensory analysis was performedwith adult individuals of commercial size (14–16 cm).These were stored in self-draining boxes and coveredwith ice (fish/ice ratio: 1:2). The boxes were stored in arefrigerated chamber (0–4 �C) and the melted ice wasreplaced daily. Anchovies were sampled immediatelyafter being caught and then every 24 h during 10 daysof storage. At each sampling time, six specimens wererandomly chosen for sensory assessment and six forfreshness chemical analysis.

Chemical composition. For the analysis of chemicalcomposition whole anchovies were used. The sampleswere homogenised with Omni Mixer, Sorvall. Watercontent was determined by drying the samples at105� 1 �C until constant weight was reached (AOAC,1995). Ash content was determined in muffle furnace at550 �C until white ashes were obtained. The proteins(%N� 6.25) were determined using the Kjeldahlmethod (AOAC, 1995). Total lipids were extractedusing the method described by Bligh and Dyer andmodified by Undeland et al. (1998). The lipid extractswere stored at �20 �C for fatty acid determination. Allanalyses were performed in triplicate. The results ofeach chemical component were expressed as g/100 gwet weight.

Fatty acid analysis. Lipids were saponified and deriva-tised to their methyl esters for fatty acid analysis

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(AOCS, 1990). The fatty acid methyl esters were ana-lysed in a Shimadzu GC-17A gas chromatograph,equipped with FID and an electronic integrator. AnOmegawax 320 fused silica capillary column(30m� 0.32mm ID, 0.25 mm phase film) was used.The oven program was 190 �C (2min), 190–225 �C(2�C/min). The injector and flame-ionisation detectorwere held at 260 �C. The carrier gas was nitrogen at aflow of 25mL/min. Identification and quantificationwere done by comparison with the retention timesand peak areas of reference standard PUFA-1,Marine Source Supelco� Supelco Inc.). Retentiontimes and peak areas were processed by Shimadzu�

SMI Class – GC 10 software

Total volatile bases nitrogen. TVB-N was determinedin acid extract of the anchovy fillet according to thereference method specified by the EU (CEE/95/149).

Lipid oxidation analysis. The lipid oxidation was deter-mined in anchovy fillet by the TBARS index, accordingto Schmedes and Holmer method (1989) and modifiedby Tironi (2005). The TBARS number was expressed asmilligram of malonaldehyde equivalents per kilogramof sample. The absorbance was determined by a spec-tophotometer (Shimadzu UV-1800) at 532 nm against ablank containing distilled water and TBARS solution.

Sensory evaluation. The methodology used to developand evaluate the QIM scheme was based on the methodearlier described by Martinsdottir et al. (2001),Sveinsdottir et al. (2003) and Hyldig and Green-Petersen (2004). In order to design the QIM, thechanges that occurred during ice storage of E. anchoitawere observed and registered by three assessors withexperience in seafood sensorial analysis. The observedchanges were considered to design the preliminary QIMscheme. Furthermore, to obtain the most appropriatedescriptors the sensory schemes available for otherEngraulidae were discussed. The QIM scheme wasdeveloped based on the changes of several parametersand its attributes such as general appearance, colourand shape of eyes, colour and odour of gills andmuscle and abdomen texture. For each attribute a setof descriptors was assigned with demerit points from 0to a maximum of 3, where 0 represented optimal qual-ity and a higher score indicated progressive deterior-ation of quality. Afterwards, training sessions wereperformed to familiarise the panel members with thedeveloped sensory scheme.

Finally, to assess the freshness and shelf-life of E.anchoita in ice storage by QIM scheme application ses-sions were performed. These sessions were carried outby eight trained panellist. Six anchovies were evaluatedby every evaluator in each of the different storage times.

The scores for the parameters were added to give thetotal sensory punctuation: the quality index (QI).Sensory characteristics were used to define the time ofrejection, based on refusal by all the assessors. This wasconsidered ideal for this species because usually, marineproducts are safe within the sensory shelf-life especiallyfor small-sized fish such as anchovies (Kose et al., 2008;Kose, 2010).

Statistical analysis. The results of chemical compos-ition and fatty acid profile of anchovy are presentedas mean value and standard deviation. Each compo-nent was analysed in triplicate. The mean values ofTVB-N and TBARS were plotted separately againstthe storage time. To establish the final QIM scheme,the sensory data were submitted to a correlation ana-lysis between the values of each parameter evaluatedand time (in days). Then, a principal component ana-lysis (PCA) was conduced to evaluate importance ofeach parameter during fish spoilage. A regression ana-lysis was carried out between the QI and the storagetime in ice. The uncertainty of prediction of days on icefrom the QI was estimated using partial least-squaresregression (PLS). The confidence level was 95% inevery statistical test used. The analyses were done byusing the statistical package STATISTICA 6.0(Statsoft, Inc., Tulsa, OK, US).

RESULTS AND DISCUSSION

Chemical composition

The chemical composition of anchovy used in thisstudy was 73.07� 1.40 g/100 g of water content,17.78� 0.34 g/100 g of protein, 7.37� 1.93 g/100 g oflipids and 2.86� 0.18 g/100 g of ash. These values thatare within the normal limits for this species (Massaet al., 2007, 2009, Garcia-Torchelsen et al., 2009), rep-resent a good substrate for microbial growth. Severalstudies show that the fat content of E. anchoita presentswide variations during the year, is minimal during thesummer months when the individuals are in post-spawning and maximal in autumn and spring wherethe spawning occurs (Yeannes and Casales, 1995).The lipids presented 31.01% of saturated fatty acidsdominated by palmitic acid (C16:0) followed by myr-istic acid (C14:0). In the monounsaturated fatty acidsfraction (41.75%), the major constituents were palmi-toleic acid (C16:1), oleic acid (C18:1) and docosaenoicacid (C22:1). The content of polyunsaturated fattyacids was 28.94%, with eicosapentaenoic acid(C20:5n-3, EPA) being the most important and doco-sahexaenoic acid (C22:6n-3, DHA). The n-3 fatty acidscontent was 24.02% of the total fatty acids. The resultsare similar to those described by Massa et al. (2007) forthis species and are in agreement with the data

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presented by other cool deep-sea fish oil such as men-haden cod sardine and anchovy (Karakoltsidis et al.,1995; Saglik and Imre, 2001; Shamsudin and Salimon,2006). The highly unsaturated fatty acids present in thisspecies, makes it very susceptible to oxidative rancidity.The importance of maintaining the nutritional qualityof this species is implicit in several studies showing thatEPA and DHA fatty acids may help to lower the risk ofchronic diseases such as heart disease, cancer and arth-ritis (Pamela, 2001; Sahena et al., 2009; Weaver andHolub, 1988). In addition, these oxidative processesmay affect sensory and technological characteristics ofthis product.

Sensory evaluation

During the preliminary sessions, the changes in sensoryparameters for E. anchoita stored in ice were examined.The parameters that presented the most significantchanges were: general appearance, eyes, colour andodour of gills, dorsal texture and abdominal aspect.Immediately after capture, the E. anchoita presented abright appearance with dark blue iridescence in dorsalsurface and a silver colour in the ventral area. Duringthe spoilage process, the skin lost its iridescence andshowed the torn muscle mainly in the abdomen(Figure 1). The eyes changed from a convex shapewith transparent cornea and black shining pupil tosunken eyes, opaque cornea and grey distorted pupil.The gills initially bright red with a characteristic ‘sea-weed’ smell changed to grey-brown colour withstrongly unpleasant odours. The opercula were silver-shiny colour at the beginning of the storage and a pro-gressive increase of bloodiness was observed duringspoilage (Figure 1). The texture was determined accord-ing to Martinsdottir et al. (2001) by finger pressing themuscle and observing how the flesh recovered. Duringthe first day of storage, the anchovies were in rigor-mortis state. After the resolution of rigor-mortis, themuscle was relaxed again and later the flesh becamesoft due to muscle autolysis and due to the action ofmicrobial enzymes (Gill 1995; Nielsen 1995). One of themain problems affecting the anchovy during storagewas bursting of the abdominal tissue (belly bursting)caused by rapid autolytic degradation. This changethat was described in other anchovies (Hernandez-Herrero et al., 2002; Pons-Sanchez Casado, 2006) isrelated to stomach content and feeding at the time ofcapture. All these parameters and their attributes weretaken into account in developing the QIM scheme.Each attribute was assigned at least three descriptorswith their appropriate score (demerit points). The finalQIM scheme developed for E. anchoita constituted of 4parameters and 10 attributes with a total of 28 demeritpoints (Table 1). All attributes of QIM scheme showed

an increasing linear with time of ice storage (R> 0.90).Other characteristics, such as mucus in surface externaland gills, were excluded by a low correlation with stor-age time (R¼ 0.58 and 0.68, respectively).

The fillet aspect was omitted in this QIM schemebecause their evaluation was laborious and destructiveto the sample. But this is an important parameter to beanalysed if products, such as marinated anchovy fillet,are manufactured. The fresh anchovy fillet was firmwith translucent and bright appearance with a red cen-treline, which originates mainly by the blood remnantfrom arteries and veins cut during the filleting. Duringstorage in ice, the flesh softens and loses its brightness.The centreline becomes brown in colour by the autoxi-dation of oxyhemoglobin into methaemoglobin. Thisfact is in accordance with Jensen (2001) and Sivertsenet al. (2009), who also mentioned that autoxidation rateof haemoglobin increases with storage temperature andvaries between species.

PCA was performed to obtain a better understand-ing of how the quality sensory parameters of E.anchoita change with time of storage in ice (Figure 2).All parameters presented highly correlated with thePCA analysis. The first component (PC1) explainedmost of the variations (93.48%) whereas the secondcomponent (PC2) contributed with 2.91%. The mostimportant parameters for PC1 were storage time(0.993), pupil of eyes (0.995), texture of dorsal muscle(0.986) and belly bursting (0.984). For PC2, cornea ofeyes (0.28) and gills smell (0.21) were parameters thatmost contributed. The parameters of quality measuredcontributed to more than 96.39% of the total informa-tion given by QIM scheme developed for E. anchoita.

The total sum of scores evaluated according to thisscheme was presented as the QI. This index linearlyincreased with the storage time in ice, which showedthat the attributes gradually deteriorated with time(Figure 3) Its evolution was represented by the equationQI¼ 2.55xþ 1.76, with x¼ days in ice; r2¼ 0.985). Theanalysis of partial least square regression (PLS) indi-cated that the regression model proposed presented astandard error of estimate¼ 1.044; p< 0.001 (Figure 4).This result indicates that the storage time could be esti-mated with an accuracy of �1 day.

Shelf-life study

To estimate the shelf-life of E. anchoita stored in ice theQIM scheme that was developed and chemical indicessuch as TVB-N and TBARS that usually are measuredto follow the spoilage pattern were used. In sensorialanalysis, the evaluators found that the anchovy remainsin excellent state during the first day of storage andretained its freshness up to the third day (QI< 12)and maintained good condition up to sixth day in ice.

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A score of 18 demerit point was considered as theacceptability sensory limit for E. anchoita in ice. Thissensory rejection point was obtained after 7 days ofstorage, mainly due to the presence of unpleasantodours and deteriorated appearance (dull pigmentationand bursting of belly). As was mentioned, the QI curveobtained by regression model can be used to predictstorage time in ice and remaining shelf-life of freshfish (Alasalvar et al., 2001; Huidobro et al., 2000;Kyrana et al., 1997).

The changes in TVB-N of anchovies stored in ice areshown in Figure 5. At the beginning of storage, TVB-Nvalue was 18.43� 0.89mg/100 g flesh. The TVB-Nvalues remained without significant differences duringthe first 5 days of storage (p> 0.05) and then started toincrease exponentially up to 30.09� 1.02mg/100 g on

day 10 of storage. The determination of TVB-N isone of the chemical indexes widely applied for theevaluation of the freshness of fish (El Marrakchiet al., 1990; Krzymien and Elias, 1990). TVB-Nvalues remained constant during autolytic degradationand then increased exponentially; this probably coin-cided with onset of spoilage and the logarithmicphase of microbial growth. This behaviour wasdescribed in several marine species stored in ice(Huss, 1995). The level of TVB-N in fish that is freshlycaught is generally between 5 and 20mg N/100 gmuscle, depending on the species, size and feedamong others factors, and levels of 30–35mg N/100 gmuscle are considered as the limit of acceptability offish stored in ice (Castro et al., 2006; Connell,1995; Huss, 1995). Although in this work TVB-N

Figure 1. General appearance of Engraulis anchoita stored in ice.

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values remained below the upper limit of acceptabilityduring the 10 days of ice storage, sensorial assessmentindicated the rejection point on day 7 of the study.

The highly unsaturated lipids present in E. anchoitaare prone to hydrolytic and oxidative reactions duringstorage and processing. This should be taken into con-sideration because in advanced stages of oxidation low-molecular-weight carbonyl compounds and alcohols

are generated, which may lead to the appearance ofobjectionable odours as well as alterations in texture,colour and nutritional value of the products(Olafsdottir et al., 1997; Sikorski 1990). Consideringthe lipid content in E. anchoita (7.37� 1.93%), it wasassumed that lipid oxidation could cause an importantimpact in the overall quality of this fish during the stor-age period. In this study, the initial value of TBARS

Table 1. Sensory quality index method scheme for Engraulis anchoita stored in ice

Parameters Attributes Descriptors Score

Generalappearance

Surfaceappearance

� Bright, dark blue iridescence in dorsal and silver in ventral surface 0

� Less bright with loss of iridescence 1

� Slightly dull, not bright 2

� Dull, the skin is removed easily from flesh 3

Skin � Complete 0

� Slightly broken 1

� Torn and damaged mainly in the abdomen 2

Operculum � Silver bright, bloodiness no visible (0%) 0

� Less bright, slight bloodiness (<10%) 1

� Some bloodiness (<50%) 2

� Very bloodiness (>50%) 3

Eyes Cornea � Clear, transparent 0

� Slightly cloudy 1

� Cloudy, opaque 2

Pupil � Bright black 0

� Dull black 1

� Grey 2

� Grey and distorted 3

Shape � Convex 0

� Flat 1

� Slightly sunken 2

� Sunken 3

Gills Colour � Bright red 0

� Dull-red 1

� Pink/light brown 2

� Discoloured grey-brown 3

Smell � Fresh, seaweedy 0

� Neutral 1

� Rancid, fishy 2

� Off odour, putrid 3

Consistency Texture � Rigid (this score is given only to fish still in rigor) 0

� Firm and elastic 1

� Slightly soft, less elastic 2

� Very soft 3

Abdomen(belly bursting)

� Intact, firm, elastic 0

� Intact, soft 1

� Slightly burst, soft 2

� Burst, very soft 3

Quality index (QI) 0–28

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was 1.66� 0.05mg malonaldehyde/kg muscles(Figure 6). This index significantly increased to1.92� 0.21mg malonaldehyde/kg on day 2 of storageand then remained without changes until the seventhday. Finally, TBARS values again showed a significantincrease reaching 2.94� 0.18mg malonaldehyde/kg atthe end of storage. It has been suggested that TBARSvalues less than 5mg malonaldehyde/kg is indicative ofgood quality of the fish, with 8mg malonaldehyde/kgbeing the limit value for consumption (Schormuller,1969, Huss, 1995). According the described results,

the limit value of TBARS for consumption was notreached in the study period. It is important to notethat according to Aubourg (1993), TBARS valuesmay not reveal the actual rate of lipid oxidation, sincemalonaldehyde may interact with other fish compo-nents. Such components may be amines, nucleosidesand nucleic acid, proteins, amino acids of phospho-lipids, and other aldehydes, which are end products oflipid oxidation. Also, this interaction varies with fishspecies. Several studies indicate that it is difficult toset limits for TBARS values without previous studies

Days in iceSkin

Operculum

TextureBelly-bursting

Eyes Cornea

Eyes Pupil

Eyes Shape

Gills colour

Gills Smell

-0.5

-0.25

0

0.25

0.5

-0.25 0 0.25 0.5 0.75 1

PC 1 (93,48 %)

PC

2 (

2,91

%)

Surface

Figure 2. Loadings in PCA of Engraulis anchoita data including all quality parameters assessed in the QIM scheme (onlythe right side of the graph is displayed).PCA: principle component analysis; QIM: quality index method.

Rej

ectio

n po

int

0

5

10

15

20

25

30

0 1 2 3 4 5 6 7 8 9 10Days in ice

QI

scor

e

Figure 3. Quality index (QI) of Engraulis anchoita in icestorage. (�) Average and standard deviation for each pointof QI. (—) Lineal correlation: Y¼ 2.5576xþ 1.7495,R2¼ 0.98; R¼ 0.992.

day 0day 1

day 2

day 3day 4

day 5day 6day 7

day 8

day day 10

0

5

10

15

20

25

30

0 5 10 15 20 25 30

Predicted values

Mea

sure

d V

alue

s

Figure 4. PLS regression model of the 28 demerit points.QIM scheme measured vs. predicted values (limits of theregression: 95% confidence).PLS: partial least squares; QIM: quality index method.

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of fat content, processing and storage conditions(Beltran and Moral, 1990; Fey and Regenstein, 1982;Liston et al., 1963).

The TVB-N index in E. anchoita during ice storageremained unchanged until fourth day and then pre-sented an exponential increase, indicating microbialgrowth. The TBARS showed a similar pattern, whichindicates an induction period in the oxidation processand subsequent increase of lipid oxidation. Althoughthese parameters show the loss of freshness of this spe-cies, they are particularly sensible after the fourth dayof storage. These results are consistent with observa-tions in other fish species (Connell, 1995; Howgate,2010c; Huss, 1995). The sensorial changes were clearlynoticeable indicating the freshness of this species all

along the stored time in ice and determining theirshelf-life.

CONCLUSIONS

The scheme sensory (QIM) developed for E. anchoitaduring ice storage was satisfactory for evaluating thefreshness and to determine its commercial shelf-life.The QI presented a linear relationship with storagetime; this suggested that QI could be used as objectivesystem of quality assessment. Under the analytical con-ditions of this study, the TVB-N value remained belowthe upper limit of marketing acceptability and TBARSvalues showed the evolution of lipid oxidations.According to the sensorial assessment the sensory rejec-tion point was on day 7 of storage.

FUNDING

We thank the Consejo Nacional de Investigaciones Cientıficasy Tecnicas (CONICET- PIP 5052), Instituto Nacional deInvestigacion y Desarrollo Pesquero (INIDEP) andUniversidad Nacional de Mar del Plata (UNMdP - 15/

G264) for their financial support.

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