Received: 4 September 2018 Revised: 8 April 2019 Accepted: 28 April 2019

DOI: 10.1002/pts.2450

RESEARCH ARTICLE

Shelf life extension of refrigerated breaded hake medallions

packed into active edible fish gelatin films

Laura M. Neira1 | Silvina P. Agustinelli2 | Roxana A. Ruseckaite1 | Josefa F. Martucci1

1
Instituto de Investigaciones en Ciencia y
Tecnología de Materiales (INTEMA), Consejo
Nacional de Investigaciones Científicas y
Técnicas (CONICET), Universidad Nacional de
Mar del Plata (UNMdP), Mar del Plata,
Argentina
2
Grupo de Investigación en Preservación y
Calidad de Alimentos (GIPCAL), Departamento
de Ingeniería Química y en Alimentos, Facultad
de Ingeniería, Universidad Nacional de Mar del
Plata (UNMdP), Mar del Plata, Argentina
Correspondence
Josefa F. Martucci, Instituto de Investigaciones
en Ciencia y Tecnología de Materiales
(INTEMA), Consejo Nacional de
Investigaciones Científicas y Técnicas
(CONICET), Universidad Nacional de Mar del
Plata (UNMdP), Av. Colón 10850, Mar del
Plata 7600, Argentina.
Email: jmartucci@fi.mdp.edu.ar
Funding information
Agencia Nacional de Promoción Científica y
Tecnológica, Grant/Award Number: PICT
2015‐2855; Agencia Nacional de Promoción
Científica y Tecnológica, Grant/Award Num-
ber: PICT 2016‐1672
Fish consumption has increased in recent years. However, fish meat is highly perish-
able, which demonstrates the need for technologies to preserve its quality. Edible
films might provide an alternative to extend the shelf life of fish. The goal of this
study was to evaluate the efficiency of edible fish gelatin films added with carvacrol
(blended: FG‐0.6CRV and grafted: FG/CRV‐g‐FG) on the quality retention of com-
mercially available prefried breaded hake medallions (BHMs). Samples of BHM were
wrapped into films, heat sealed, and stored at −4 ± 1 °C for 49 days. Free carvacrol
gelatin films and the unwrapped medallions were used as controls. After
predetermined periods of time, samples were analysed in terms of microbiological
and physicochemical properties. Overall the storage period, the total volatile basic
nitrogen value (TVBN) and the total psychrotrophic bacteria counts (TPC) were signif-
icantly lesser in BHM wrapped in FG‐0.6CRV than in the other samples. The total oxi-
dation index (TOI), peroxide value (PV), and p‐anisidine value (AV) were reduced by
18%, 30%, and 16%, respectively, compared with controls. FG/CRV‐g‐FG film was
not efficient to reduce the microbial spoilage or the oxidation, probably due to the
retention of CRV into the matrix avoiding its migration towards the foodstuff, proving
the importance of checking the films functionality in real food systems. Sensory eval-
uation of fried samples wrapped in FG‐0.6CRV was also performed. The results indi-
cate that the medallions can be directly cooked and consumed without removing the
packaging with a good acceptance by the consumer.

K E Y W OR D S

active films, breaded hake medallions, carvacrol, fish gelatin, shelf life

1 | I N T RO D U CT I O N require a minimal final cooking (usually deep frying) by the consumers

Fish, an excellent low‐fat source of protein, provides many health
benefits, such as omega‐3 fatty acids that reduce cholesterol levels,
the incidence of stroke, heart disease, preterm delivery, and enhances
cognitive development.1 According to the elaboration and preserva-
tion methods, fishery, meats, and dairy products are similar in nutri-
tional quality.2 In order to make fish products more attractive, the
battering and breading techniques have been extensively used to
improve the flavour and texture of fish.3-5 Fish nuggets and medal-
lions are prepared (battering coat and prefried) products that only
or food service providers.6 Without the presence of a preservative
compound, these kind of products are susceptible to physical, chem-
ical, and microbiological spoilage. The first two types of spoilage have
a negative impact on sensory attributes, while the latter leads to
health and economic issues.7
Natural antioxidant and antimicrobial agents are widely used for
food preservation. Many studies have been focused on the antioxi-
dant and antimicrobial agents present in essential oils (EOs) extracted
from plants (basil, thyme, oregano, cinnamon, clove, rosemary)
consisting of complex mixtures of different compounds including

Packag Technol Sci. 2019;1–10. wileyonlinelibrary.com/journal/pts © 2019 John Wiley & Sons, Ltd. 1
2 NEIRA
terpenoids, esters, aldehydes, ketones, acids, and alcohols.8 Extracts
derived from herbs and EOs contain many natural compounds such
as thymol, linalool, and carvacrol with a broad antimicrobial range
against different pathogenic and spoilage microorganisms including
gram‐negative and gram‐positive bacteria,9,10 as well as against
yeast11 and molds.12 In general, EOs are considered to be safe, and
they have been classified as generally recognized as safe (GRAS) by
13-18
the American Food and Drug Administration. The structure and
characteristics of EOs and active compounds19 and their prospective
application in meat and bakery industries have been extensively stud-
ied.5,20-25 One interesting alternative to extend the shelf life of fish
breading products is the incorporation of these natural additives to
edible films. Edible film can be prepared from natural polymers such
13,26
as proteins, polysaccharides, and lipids. As a typical protein film‐
based material, gelatin is used due to its abundance, acceptability,
low manufacturing cost, and great film‐forming properties. 13,26,27
There are several works in the literature, where active protein films
are used for the preservation of fish products with promising
results.13,14,28-33 Ahmad et al34 reported the extension of shelf life
of sea bass slices wrapped in fish gelatin (FG) films incorporated with
cinnamon EO during storage at 4 °C for 12 days, mainly by retarding
the microbial growth and lowering the lipid oxidation. Kaewprachu
et al35 investigated the changes in various distintic attributes of refrig-
erated bluefin tuna slices wrapped with active fish myofibrillar protein
(FMP) films for 10 days. The microbiological, chemical, and sensory
evaluation showed an increase of 400% of shelf life of the tuna
wrapped with FMP. Similar results were obtained by other authors in
pigskin gelatin films with oregano and rosemary extracts for the con-
servation of cold‐smoked sardine, 33
in gelatin–chitosan film containing
clove EO to wrapp refrigerates cod fillets,36 in gelatin–chitosan film
containing oregano EO for fresh grass carp muscle preservation37
and in refrigerated rainbow trout slice covered with gelatin–alginate
film containing oregano EO.38
The active agent is generally incorporated by blending with the bio-
polymer matrix, and therefore, it has enough mobility to diffuse towards
the food with time. The main drawbacks of this method are the incorpo-
ration of odour and flavour to the packed food and the alteration of
mechanical and functional properties of films during time.39 To over-
come the above mentioned limitation, a useful approach is the covalent
linkage of active substances onto polymer matrix, enhancing their sta-
9,39-44
bility and reducing the effects of migration. Phenolic substances
were conjugated onto polysaccharides43,45 and proteins9,42 to obtain
antimicrobial and/or antioxidant materials with potential application
for food preservation. To ensure the viability of the material to be used
as a food packaging, it is necessary to carry out in vivo tests where the
material interacts with the food. Schreiber and coworkers39 obtained
gallic acid‐grafted chitosan films (GA‐g‐CH) and showed the efficiency
of GA‐g‐CH for conservation of unroasted peanuts at 50 °C for
15 weeks.39 To our acknowledged, the bibliography about the use of
grafted biopolymer films in contact with food is scarce.
The objective of this study was to determine efficiency of grafted
FG films with carvacrol (FG/CRV‐g‐FG) as packaging material for com-
mercially available prefried breaded hake medallions (BHMs) in
ET AL.
comparison to FG films added with carvacrol by bleding (FG‐
0.6CRV). Changes in microbiological quality (mesophilic and
psychrotrophic bacteria) and in physicochemical parameters (pH,
humidity retention (HR), total volatile basic nitrogen (TVBN), peroxide
value (PV), and p‐anisidine value (AV)) during storage at −4 ± 1 °C
were evaluated. To assess the effectiveness of the active film in a
packaging system for ready to eat BHM, sensory evaluation of
wrapped and fried BHMs pieces was performed.
2 | MATERIALS AND METHODS
2.1 | Materials
The FG was kindly supplied by Rousselot (Argentina), bloom strength
208, and isoionic point (Ip) 8.6. Buffer Phosphate pH 7, hydrogen per-
oxide (H2O2), and methanol were obtained from Anedra (Argentina).
5‐Isopropyl‐2‐methylphenol (CRV, 98%), 1, 1‐Diphenyl‐2‐
picrylhydrazyl (DPPH), peptone, potassium iodide, p‐anisidine, and
ascorbic acid were from Sigma Aldrich (USA). Glycerol (Gly, 98%)
was supplied by DEM Chemicals (Argentina). Boric acid, acetic acid,
chloroform (99%), and sulphuric acid (98%) were purchased from
Biopack (Argentina). Sodium chloride and magnesium oxide were from
Cicarelli (Argentina). Sodium thiosulfate and isooctane (p.a.) were
obtained from Merck (Argentina). Plate Count Agar (PCA) was from
Britania (Argentina). All reagents were analytical grade and used as
received.
Prefried BHMs (Hake, Merluccius hubbsi) were supplied by Grangy's
Frozen Food, (Mar del Plata), proximate composition (on wet basis):
18.46% carbohydrates; 13.84% proteins; 6.92% total fats; 0.61% die-
tary fibre, and 0.32% sodium.
2.2 | Methods
2.2.1 | Chemical modification of FG
CRV was grafted onto FG powder following the procedure described by
Spizzirri et al,44 with some modifications. FG was dissolved in distilled
water (1% w/v) at room temperature under constant stirring. Then,
1 mL of H2O2 (100 vol) and 0.25 g of ascorbic acid were introduced into
FG solution and mixed at 25 °C for 2 hours. At that point, CRV
(0.35 mM) was added to the solution and let to react for 24 hours under
nitrogen atmosphere. The solution was dialyzed (MWCO: 12‐14,000)
against distilled water for 48 hours with two changes of water per
day. The presence of unreacted CRV was detected by colour test with
FeCl3 solution (1% w/w). The dialysed solution was freeze‐dried (Virtis
Benchtop SLC lyophilizer, USA) and the carvacrol‐grafted FG (CRV‐g‐
FG) was obtained as a white powder.9
2.2.2 | Film preparation
Active FG films were prepared by two methods: (a) by blending carva-
crol with FG and (b) by blending FG with the appropriate amount of
NEIRA ET AL.
CRV‐g‐FG in order to attain the same antioxidant activity as deter-
mined by radical scavenging activity against DPPH.9 FG (10 g) was dis-
solved into distilled water (100 mL), and glycerol (2 g) was added as
plasticizer. The mixture was stirred for 30 minutes at 40 °C and
500 rpm to achieve a film‐forming solution (FFS). Carvacrol (0.6%
w/v) was incorporated into the FFS and homogenized using a homog-
enizer (Ultra Turrax IKA T25, Germany) at 11,000 rpm for 1 minute
and 20,000 rpm for another minute. This concentration is sufficient
to ensure antioxidant and antimicrobial activity in the film without
imparting taste or odour to the packed food.9 The obtained FFSs were
poured into Petri dishes with Teflon and dried at 40°C until constant
weight to obtain films (FG‐0.6CRV). The CRV concentration in the
resulting film was 0.591% w/v,10 indicating a retention of the 98.5%
of CRV after processing. Control films (FG‐0CRV) were achieved with
the same procedure without CRV.
In the method (b), FG (5 g), CRV‐g‐FG (5 g), and glycerol (2 g) were
dissolved together into distilled water (100 mL) and stirred for
30 minutes (40 °C, 500 rpm). The obtained FFS was poured into Petri
dishes coated with Teflon and dried at 40 °C until constant weight to
obtain films (FG/CRV‐g‐FG). All films were peeled off the molds and
stored at 23 ± 2 °C and 65 ± 5% RH, during 2 days before testing.
2.2.3 | Wrapping BHM into active FG films
Ultra frozen BHMs (dimensions: 8.5 cm in diamter, 1 cm in thickness,
weight 87 ± 1 g) were separated into four lots: (a) unwrapped medal-
lions (control), (b) wrapped into FG film without CRV (FG‐0CRV), (c)
wrapped into FG‐0.6CRV, and (d) wrapped into FG/CRV‐g‐FG films.
Edible pouches were made by stacking together two lays of the same
film formulation (15 cm in diameter) and heat sealed by using a manual
tray sealer (Envamar, Argentina) with temperature control (220 V,
400 W, and 1 second). Each pouch was filled with one medallion
(87 g). Batches were stored at −4 ± 1 °C for 49 days. Samples were
taken at predetermined times and evaluated for microbiological and
physicochemical properties. Three samples of each group were
analysed at each sampling time.
2.2.4 | Total aerobic plate count assay
The BHM (10 g) were aseptically homogenized for 1 minute using a
Stomacher 400 Circulator Homogenizer laboratory blender (Seward,
UK) with 90 mL of diluent solution (8.5 g of NaCl/L, 1 g of
46
peptone/L, and 1 mL each 800 mL of phosphate solution [pH 7).
Decimal dilutions of the homogenate were prepared in peptone saline
solution by triplicate according to the recommended microbiological
protocols.46 Samples of serial dilutions were spread on the surface
of Petri dishes for determination of total aerobic plate count (TAPC),
on PCA, and incubated at 35 ± 0.5 °C for 2 days (mesophilic bacteria
count, MTC) and at 7 ± 0.5 °C for 10 days (PTC bacteria). Counted col-
onies were reported as the logarithm of an average of three replicates
per gram (log CFUs/g, CFU: colony‐forming units).
3
2.2.5 | Physicochemical analysis during the storage
period
Variations in HR, pH, TVBN, TOI, PV, and AV were followed over the
whole period of storage.
1. HR was determined by oven drying the samples at 105 ± 2 °C
(Mermmet, Germany) until constant weight according to the
method described in Association of Official Analytical Chemists
(A.O.A.C).47 Results were the average of three measurements for
each sample.
2. pH variation was analysed using a portable pH meter (Hanna, USA)
with HI1271 pH electrode; 10 g of minced BHM was homogenized
with 10 mL of distilled water, let to stand at room temperature,
and then, the pH was tested. Reported values were the average
of three replicates.
3. TVBN was determined by distillation method.48 A precise amount of
minced medallion (10 g) was placed into a distilling flask and blended
with 300 mL of distilled water and 1.5 g of MgO. After steam distil-
lation, volatile bases were absorbed by boric acid solution (3% w/v)
until completing a volume of 230 mL in the beaker and titrated with
a normalized sulphuric acid solution. TVBN content was calculated
by triplicate and expressed as mg TVBN/100 g of sample.
4. PV of lipid extracted from BHM was determined by iodometric titra-
tion according to a standard procedure indicated in the Official
Methods and Recommended Practices of the American Oil Chem-
ists' Society (AOCS) .49 A lipid sample (1 g) was dissolved in 30 mL
of the 3:2 acetic acid: chloroform solution, followed by the addition
of 0.5 mL of saturated KI solution and manual shaking for 1 minute.
After the addition of 30 mL distilled water, the sample was titrated
against sodium thiosulfate solution. Oil was extracted by the
method proposed by Bligh and Dyer.50 All determinations were
made in duplicate. The results were expressed as milliequivalents
(meq) of active oxygen per kilogram of lipid (meq O2/kg).
5. AV was determined by the standard method described in AOCS 51
using a spectrophotometer UV–visible Shimadzu 1601 PC (Tokyo,
Japan) at 350 nm. The data are representative of two oxidation
experiments performed by duplicate.
6. TOI was calculated from the relationship52:
TOI ¼ 2PV þ AV (1)
2.2.6 | Sensory evaluation
Sensory evaluation was performed by 30 untrained panellists of differ-
ent ages (20‐60 years), sex (45% male and 55% female), and occupa-
tions, to validate the test.53 Only samples of lot B (packed into FG‐
0CRV, control film) and C (packed into FG‐0.6CRV) were tested.
BHMs were aseptically cut with a punch (2 cm in diameter), packed
4 NEIRA
into edible pouches (4 × 4 cm), and heat sealed. Before the evaluation,
all wrapped samples were fried in vegetable oil at 180 °C for
3 minutes,9 followed by drying with absorbent paper to remove the
oil excess. Samples of fried wrapped BHMs, coded with three‐digit
random numbers, were given to the panellists asked to rate the global
acceptance of the different batches by judging the aroma, colour,
crunch, taste, cooking degree, and global acceptability on a hedonic
scale of 7 points (7 = like immensely, 6 = like very much, 5 = like
slightly, 4 = neither like or dislike, 3 = dislike slightly, 2 = dislike very
much, 1 = dislike immensely).54 Tests were carried out in individual
cabins for sensory analysis studies. Coffee and water were used to
neutralize aroma and flavour, respectively, between each sample eval-
uated. The sequence of preparation, frying, and testing is represented
in Figure 1.
2.2.7 | Statistical analysis
Data values obtained in the experiments were statistically analysed by
one‐way analysis of variance (ANOVA) employing OriginLab 8 soft-
ware. Differences between pairs of means were assessed on the basis
of confidence intervals using the Tukey test. The least significant dif-
ference was p <0.05.
3 | RESULTS AND DISCUSSION
3.1 | Properties of gelatin–CRV film
In our previous study, oregano EO (OEO) and their dominant com-
pound (carvacrol, CRV) were used to impart antioxidant and antimicro-
bial activity to gelatin films.8,10 The incorporation of 0.6% of these
additives induce satistically the same antioxidant capacity
(%RSA = 56.2 ± 1.2 and 60 ± 4% for gelatin‐0.6CRV and gelatin‐0.6
OEO, respectively), but the CRV incorporation produces the best
FIGURE 1 Preparation of samples for sensory assessment. (A) Cutting BHM (2 × 2 cm), (B) wrapping and heat sealing of BHM samples, (C) frying
process (180 °C and 3 minutes), (D) fried BHM. (E) The samples were introduce into the testing boxes (F) in individual containers, and (G, H) were
test by an untrained evaluator
ET AL.
antimicrobial performance of films (Inhibition halo against Escherichia
coli = 31.0 ± 1.0 and 18.0 ± 0.8 mm for gelatin‐0.6CRV and gelatin‐
0.6OEO, respectively).8,10 For this reason, carvacrol‐added gelatin
films were selected as potential protective films for food product.
The most important properties related with the preservative effects
of the film on food are their antimicrobial and antioxidant capacity,
water vapour (WVP) and oxygen permeability (OP). The incorporation
of CRV by blending (FG‐0.6CRV) or grafting (FG/CRV‐g‐FG) intro-
duced antioxidant capacity (%RSA = 56.2 ± 1.2, 60.1 ± 2.2 and
4.1 ± 1.1%/g for FG‐0.6CRV, FG/CRV‐g‐FG, and FG, respectively),
antibacterial properties (inhibition halo against Escherichia
coli = 31.0 ± 1.0, 8.0 ± 0.2, and 0 ± 0 mm for FG‐0.6CRV, FG/CRV‐
g‐FG, and FG, respectively), and improved water vapour permeability
of FG‐films (WVPFG‐0.6CRV = 3.5 ± 0.7 × 10−9; WVPFG/CRV‐g‐
FG = 2.1 ± 0.2 × 10−9 vs WVPFG = 8.8 ± 0.7 × 10−9 g/Pa.h.cm), without
altering the mechanical properties, optical attributes, and OP (OP=
2.8 ± 0.6, 3.2 ± 0.3 and 2.5 ± 0.5 cm3.μm.m−2.d−1.kPa−1 for FG‐
0.6CRV, FG/CRV‐g‐FG, and FG, respectively).9
3.2 | Aerobic plate count
The total microbial load of BHMs depends on the food ingredients and
the sanitary conditions as well as the storage time and temperature.
Figure 2 shows the variation of MTC in BHMs during refrigeration.
The initial MTC of BHM samples (before testing) was 3.1 log CFU/g,
in accordance with the data reported from fish finger immediately
after preparation.55,56 The MTC of unpacked control samples (lot A)
and packed in control film (FG‐0CRV, lot B) steadily increased up to
the 28th day. Beyond that time, lot A attained a maximum aerobic bac-
terial count close to 4.2 log CFU/g, which was below 7 log CFU/g that
is considered the onset of spoilage of fish products according to
FAO57 and the International Commission on Microbiological Specifica-
tions for Foods (ICMSF).56 Samples packed in FG‐0.6CRV (lot C) and
NEIRA ET AL. 5

the surface of the packed food, resulting the most effective against
PTC bacteria.62 In samples packed in FG/CRV‐g‐FG (lot D), the antimi-
crobial activity is just induced by the direct contact with the product
because the active agent is covalently attached (immobilized) to the
protein matrix.
The reduction in PTC bacteria of fisheries products by packaging
with protein active films compared with unwrapped and control films
was previously reported.28,34,35 Ahmad and co‐workers34 evidenced
the reduction of PTC in slices of refrigerated sea bass as a conse-
quence of packaging with gelatin films incorporated with lemongrass
vs control films. Andevari and Rezaei29 indicated that a gelatin coating
enriched with cinnamon oil is suitable for extending the shelf life of
fresh rainbow trout fillets while maintaining an acceptable quality dur-
ing storage¸ the active film reduced total bacteria growth during
15 days of cold storage compared to control films.

FIGURE 2 Total aerobic mesophilic microorganism (solid line) and 3.3 | Physicochemical analysis
psychrotrophic bacteria (dash line) colonies counting in refrigerated
breaded hake medallions (−4 ± 1 °C) as a function of the conservation
3.3.1 | Humidity retention
time and treatment: (■) lot a (control), (●) lot B (FG‐0CRV), (▼) lot C
(FG‐0.6CRV), and (▲) lot D (FG/CRV‐g‐FG)
Figure 3 shows the evolution of the HR during storage at −4 ± 1 °C.
Lot A showed a sustained drop in the water content that became sig-
FG/CRV‐g‐FG (lot D) films did not show colonies of mesophilic bacte- nificant after 20 days (p < 0.05). All packaged samples (lots B, C, and D)
ria (p < 0.05, Figure 2) from the seventh day of preservation until the retained a significantly higher percentage of water (p < 0.05). Thus, the
end of the study, demonstrating the ability of such films to delay the FG‐based films act as additional barriers to prevent moisture loss from
microbial growth. Since FG has no significant antibacterial activity,10 packaged samples, in spite of WVP values of FG‐based films (section
the inhibitory effect was ascribed to the presence of carvacrol, either 3.1.). No significant differences (Figure 3, P > .05) in humidity content
8,58
blended or grafted, in both packaging formulations. Carvacrol has a of lots B and C were detected, suggesting that the inclusion of 0.6%
well‐known broad‐spectrum antimicrobial activity against gram‐ CRV did not modify the moisture transfer through the protein film
positive and gram‐negative bacteria, fungi, molds, and yeasts.59 The under the preservation conditions analysed (T conservation <Tg ~49
antibacterial effectiveness of CRV lays in its phenolic structure and °C,9). The unfavourable effect observed in samples of lot D (BHM
60
hydrophobic character that allow it to penetrate through the cell packed in FG/CRV‐g‐FG film) was attributed to the presence of
wall of a bacterium and attack on the phospholipid bilayer present in
cell. This increases the membrane fluidity and favours the leakage of
protons and potassium ions, leading to a decrease in the pH gradient
across the cytoplasmic membrane, a collapse of the membrane poten-
tial, the inhibition of ATP synthesis, and finally, the cell death.60 Pro-
tein edible films containing oregano or thyme EOs as well as their
primary active components (carvacrol and thymol) led to a decrease
in the microbial growth on refrigerated cod,36 tuna fillets,32 and rain-
bow trout slices.38 Reduction in total bacterial count of braded fish
fingers coated with chitosan films upon storage at −18 °C for 6 month
was reported with TBC lower than 4 log CFU/g at the end of the stor-
age period.
The PTC bacteria are the primary group of microorganisms respon-
sible for the spoilage of aerobically stored fresh fish at low tempera-
tures.61 In our study, the initial count (day 0) in BHM was 2 log CFU/g
(Figure 2). PTC gradually increased with storage time and samples of
lots A, B, and D behaved similarly (Figure 2). Contrarily, at the end
FIGURE 3 Humidity retention of the refrigerated products (−4 ± 1
of the experiment, lot C samples showed a significantly lower count
°C) during time and treatment: (■) lot a (control), (●) lot B (FG‐
(4 log CFU/g) compared with values observed for the other samples
0CRV), (▼) lot C (FG‐0.6CRV), and (▲) lot D (FG/CRV‐g‐FG). Inset:
(5‐6 log CFU/g). In film used in lot C, the active is not covalently optical microscopy (50×) of the surface of FG/CRV‐g‐FG film after
attached to the matrix, and therefore, it is freely to diffuse towards 14 days of storage
6 NEIRA ET AL.

microcracks and pits that were evident after 14 day of storage by opti-
cal microscopy of the surface of the film (inset in Figure 3). Consistent
with these results, Nilsuwan, Benjakul, and Prodpran63 reported that
the presence of FG films as a protective package for dry products such
as cookies induce a reduction in water absorption.
3.3.2 | pH
The initial pH value of unpacked BHM was 6.41 ± 0.04 (Figure 4),64
and it increased up to 7.32 after 28 days of refrigeration, near to the
limit value for the safe consumption of this type of products.65 This
increment of pH reflects the accumulation of alkaline compounds,
mainly derived from microbial fish muscle spoilage.66 For all the pack-
aged products, the pH values varied between 6.66 and 7.09 (Figure 4),
and no significant differences (p > 0.05, Figure 4) were detected
between samples wrapped into FG‐0CRV and FG/CRV‐g‐FG (lot B
and D, respectively) during the whole conservation period in agree-
ment with the development of total PTC bacteria in the product
(p < 0.05, Figure 2). Samples of lot C showed the lowest values of
pH during all storage period. The slight initial pH increment observed
for samples wrapped in FG‐0.6CRV was associated with the delay in
the microbial degradation processes because of the antimicrobial
action of CRV (p > 0.05, Figure 2). Ahmad et al34 analysed the pres-
ervation of sea bass slices wrapped in FG films with lemongrass oil
at 4 °C and recorded a lower increase in the pH of the wrapped
product after 12 days of conservation, being the most noticeable
31
effect for the active film. Chandra Mohan and co‐workers
reported that the incorporation of natural extracts to films based
on tamarind seed starch for the conservation of seer fishes
(Scomberomorus guttatus) at 4 °C allowed maintaining the pH of
the packaged product during a longer storage time, associated with
the antimicrobial properties of the container that indirectly reduce
protein deterioration.31

3.3.3 | Total Volatile Basic Nitrogen, peroxide value,

anisidine value, and total oxidation index

The measure of TVBN is directly related to the growth of microorgan-

isms and the formation of basic compounds that result from their
metabolism and that lead to an increase in pH.48,67 As the storage time

FIGURE 4 pH values of the breaded hake medallions samples during increased, the TVBN values of packed and unpacked samples continu-
the conservation time at −4 ± 1 °C. (■) lot a (control), (●) lot B (FG‐ ously increased, but the increasing rate varied amongst samples. Con-
0CRV), (▼) lot C (FG‐0.6CRV), and (▲) lot D (FG/CRV‐g‐FG) trol samples (lot A) showed a rapid increase in TVBN, from 15 mg of

TABLE 1 Total volatile basic nitrogen (TVBN), peroxide value (PV), p‐anisidine value (AV) and total oxidation index (TOI) of the samples during
the conservation period

Sample Days TVBN, mg/100 g PV, meq O2/kg of lipid) AV TOI (2PV + AV)

Control (lot A) 0 1.50 ± 0.17a 1.62 ± 0.14a 20.12 ± 1.10a 23.36 ± 1.38a
7 1.51 ± 0.03a 3.93 ± 0.21b 27.50 ± 0.37b 35.36 ± 0.79b
14 1.86 ± 0.08b 3.42 ± 0.08c 33.40 ± 0.85c 40.24 ± 1.01c
28 2.49 ± 0.03c 3.35 ± 0.12c 37.86 ± 1.20d 44.56 ± 1.44d
49 2.90 ± 0.08d 3.24 ± 0.11c 55.76 ± 3.21e 62.24 ± 3.43e
FG‐0CRV (lot B) 0 1.50 ± 0.17a 1.62 ± 0.14 a 20.12 ± 1.10a 23.36 ± 1.38a
7 1.57 ± 0.07a 3.14 ± 0.25c 25.24 ± 2.28b 31.52 ± 2.78b
14 1.62 ± 0.11a 2.64 ± 0.16d 29.89 ± 1.95bf 35.17 ± 2.27bc
28 2.21 ± 0.02e 2.68 ± 0.08d 33.45 ± 3.65cf 38.81 ± 3.81c
49 2.52 ± 0.06c 2.72 ± 0.26d 48.87 ± 2.56g 54.31 ± 4.02d
FG‐0.6CRV (lot C) 0 1.50 ± 0.17a 1.62 ± 0.14a 20.12 ± 1.10a 23.36 ± 1.38a
7 1.67±0.00a 2.69 ± 0.23de 22.33 ± 0.82a 27.71 ± 1.28a
14 1.73 ± 0.11a 2.56 ± 0.06de 28.68 ± 1.56b 33.80 ± 1.68b
28 1.87 ± 0.08b 2.45 ± 0.21de 31.56 ± 0.95f 36.46 ± 1.37c
49 1.97 ± 0.09b 2.36 ± 0.10e 46.64 ± 3.01g 51.36 ± 3.21d
FG/CRV‐g‐FG (lot D) 0 1.50 ± 0.17a 1.62 ± 0.14a 20.12 ± 1.10a 23.36 ± 1.38a
7 1.56 ± 0.10a 2.96 ± 0.28c 28.95 ± 1.02b 34.87 ± 1.58b
28 2.37 ± 0.11ce 3.25 ± 0.12c 34.37 ± 2.36cd 40.87 ± 2.60c
49 2.66 ± 0.00d 3.57 ± 0.41c 55.16 ± 2.25e 62.30 ± 3.07d

Note. Equal letters in the same column indicate that there is no significant difference between the results. Tukey test, 95% confidence
NEIRA ET AL. 7

TVBN/100 g of sample up to 29 mg of NBVT/100 g sample at the end
of the 49 days of storage (Table 1) while the lower increasing rate was
observed for lot C samples, wrapped in FG‐0.6CRV film (Table 1).
TVBN contents were in accordance with the microbial load detected
during storage in the corresponding samples (Figure 2). As a general
trend, all stored samples kept TVBN content below 30 to 35 mg of
TVBN/100 g sample that is the threshold limit for fresh fish according
to the European Commission.67 The reduction in TVBN values of films
based on biopolymers incorporated natural extracts and EOs have
demonstrated that these films constrain the microbial growth in fish-
ery products.31,34
The evaluation of lipid oxidation over packaged product was
determined by the PV, and the AV, as indicators of the primary
and secondary oxidation products, respectively.49,51 The initial PV
of BHM samples was 1.62 ± 0.14 meq O2/Kg (Table 1), similar to
values reported for frozen mackerel burgers (3.6 meq O2/Kg).68
The low PV value was associated with the small fat content of fish
TABLE 2 Attribute scores of sensory evaluation of breaded hake
medallions wrapped in active (FG‐06CRV) and control (FG‐0CRV) films
Evaluated attribute FG‐0CRV FG‐0.6CRV
species used for the medallions and the good quality of raw material.
Samples without film (lot A) showed the highest PV at all sampling
points (Table 1), thus showing the protective effect of protein film
associated with its low OP,27 irrespective of the presence of the
active compound (section 3.1).9 In the same way, other authors indi-
cated that protein‐based coatings delayed lipid oxidation of salmon
fillets measured by PVs and TBARS.69 The PV values of HBMs
wrapped with FG/CRV‐g‐FG (lot D) increased continuously during
the whole storage period up to values similar to those of samples
packed in GP‐0CRV. In addition, the CRV/CRV‐g‐FG showed the
highest PI values. The presence of microcracks in FG/CRV‐g‐FG film
(Figure 3) and the higher superficial hydrophobicity caused by
grafting CRV onto FG surface (surface polarity (%), 7.92 ± 0.01 vs
4.34 ± 0.06, for FG‐0.6CRV and FG/CRV‐g‐FG, respectively) might
account for the increased PV and PI values as compared with FG‐
0.6CRV. The samples of lot C (packed into FG‐0.6CRV) displayed a
reduction in PV of about 30% (PV = 2.36 meq O2/Kg), more likely
due to the antioxidant ability of CRV,69 since OP values of both
films were statistically similar (section 3.1.).9
Similar results were reported in “seer fishes” packed in films of tam-
arind seed starch with natural extracts after 20 days of storage at 4 °C,31

Aroma 5.18 ± 1.05a 4.85 ± 1.21a in rainbow trout fillets wrapped with gelatin containing laurel EO28 and
in tuna slices wrapped with FMP film incorporated with catechin‐
Colour 4.57 ± 1.07a 4.64 ± 1.31a
Kradon extract.35 Giménez and co‐workers68 informed a 50% reduction
Flavour 5.61 ± 0.87a 5.25 ± 0.89a
in the PV relative to the control, after 60 days of storage of mackerel
Crunchiness in mouth 5.28 ± 1.24a 5.36 ± 1.25a
burgers packed in gelatin films with borage extract (ratio 1:1).
Cooking degree 5.21 ± 1.23a 5.39 ± 1.16a The AV and TOI values evolved in a similar manner than PV upon
Overall acceptability 5.25 ± 0.93a 5.00 ± 1.41a storage (Table 1). Samples of B and C lots exhibited lower AV and TOI
Note. Equal letters in the same row indicate that there is no significant dif- values than the control (lot A) and ample wrapped in FG/CRV‐g‐FG
ference between the results. Tukey test, 95% confidence during the whole conservation period, indicating a delay in the

FIGURE 5 Overall acceptability of fried

breaded hake medallions packed into FG‐
0CRV (control) and FG‐0.6CRV films
8 NEIRA
rancidity process of the product associated with the good oxygen bar-
rier of FG‐based films.10 FG‐0.6CRV film (lot C) was the most effective
against the oxidation because the combination of the inherent oxygen
barrier property of FG film and the presence of CRV blended in the
matrix (AV: 46.64 ± 3.01 vs 55.76 ± 3.21, TOI: 51.36 ± 3.21 vs
62.24 ± 3.43, for lot C and A after 49 days of conservation, respec-
tively).70 Unlike samples of groups B and C, BHMs packed in
FG/CRV‐g‐FG film, did not exert any protection against lipid oxidation
arising from the inability of CRV to migrate towards the food surface
and the poor oxygen barrier because of microscopic.
Schreiber et al39 developed chitosan films grafted with Gallic acid
and applied them for the storage of crushed peanuts for 15 weeks
and French fries for 8 weeks at 50 °C. The authors observed a signif-
icant reduction in the PV and in thiobarbituric acid reactive substances
between the samples packed in the grafted film and the control sam-
ples, demonstrating the antioxidant effectiveness of the packaging.39
3.4 | Sensory assessment of fried BHM wrapped in
FG films
Amongst the analysed films, FG‐0.6CRV showed the best results in
BHM conservation under the conditions studied. Accordingly, the
acceptance test was carried out on fried BHM packed into FG‐0CRV
(control) and FG‐0.6CRV, and results are summarized in Table 2. Sam-
ples wrapped in the active film were broadly accepted as revealed by
the statistically similar scores of all the sensory attributes evaluated,31
as compared to the control (p > 0.05, Table 2). The panellist expressed
their negative opinion related to the sensory characteristics of the
BHM packed in FG‐0.6CRV, more likely due to the slightly invasive
aroma and flavour of CRV in the fried product. The presence of CRV
in edible film reduce the percentage of acceptability of cooked BHM
(Figure 5, 75 vs 82% with score > 5),31 but the values are still elevated.
Our results are in line with those reported for others. Du et al58 pro-
duced tomato‐based films with and without carvacrol for the preser-
vation and cooking of chicken. The addition of 0.5% CRV did not
affect the preference of the consumers of the chicken cooked with
any of the analysed films; however, an increase to 0.75% CRV drasti-
cally reduced the preference because of the aroma and flavour
imparted by the CRV to the product. Likewise, it was demonstrated
that the incorporation of oregano oil (whose main constituent is carva-
crol) reduced the acceptability of meat products by consumers.71
4 | C O N CL U S I O N S
Considering the health tendency of the modern food technology, the
use of bio‐based active packaging is highly promising for the future
improvement of food quality and preservation during processing
and storage of different food materials. This study proved that FG
films incorporated with 0.6% carvacrol by blending FG‐0.6CRV
delayed the chemical and biological spoilage of prefried BHM stored
at −4 ± 1 °C for 49 days. Results revealed the relevance of migration
of carvacrol as the active agent towards the packed food to impart
ET AL.
its antioxidant and antimicrobial protective action. It must be
highlighted that the FG‐0.6CRV active packaging materials can
be useful for preserving and extending the shelf life of prefried
BHM. Moreover, FG‐0.6CRV is a potential active edible packaging
because all the components are non‐toxic and GRAS, with the
advantage that the packed food can be directly cooked and con-
sumed without removing the packaging with a good acceptance by
the consumer.
ACKNOWLEDGEMENTS
Authors would like to express their gratitude to Consejo Nacional de
Investigaciones Científicas y Técnicas (CONICET) and Agencia
Nacional de Promoción Científica y Tecnológica (ANPCyT, grant num-
ber PICT 2016–1672 and PICT 2015‐2855) of Argentina for their
financial support.
ORCID
Roxana A. Ruseckaite https://orcid.org/0000-0002-7409-5546
Josefa F. Martucci https://orcid.org/0000-0002-9789-4359
RE FE RE NC ES
1. Willett WC. Fish: balancing health risks and benefits. Am J Prev Med.
2005;29(4):320‐321.
2. Maas‐van Berkel, B., van den Boogaard, B., & Heijnen, C. Preservation
of fish and meat. 2004. Agromisa Foundation, Wageningen, Editor:
Marja de Goffau‐Markusse. ISBN: 90–72746–01‐9
3. Biswas S, Chakraborty A, Sarkar S. Comparison among the qualities of
patties prepared from chicken broiler, spent hen and duck meats. J
Poult Sci. 2006;43(2):180‐186.
4. Duman M, Kuzgun NK. Quality changes of nugget prepared from fresh
and smoked rainbow trout during chilled storage. Br Food J.
2018;120(9):2080‐2087.
5. Gavahian M, Chu Y‐H, Lorenzo JM, Khaneghah AM, Barba FJ. Essential
oils as natural preservatives for bakery products: understanding the
mechanisms of action, recent findings, and applications. Crit Rev Food
Sci Nutr. 2018;1‐12. https://doi.org/10.1080/10408398.2018.
1525601
6. Albert A, Perez‐Munuera I, Quiles A, Salvador A, Fiszman SM,
Hernando I. Adhesion in fried battered nuggets: performance of differ-
ent hydrocolloids as products using three cooking procedures. Food
Hydrocoll. 2009;23(5):1443‐1448.
7. Smith JP, Daifas DP, El‐Khoury W, Koukoutsis J, El‐ Khoury A. Shelf
life and safety concerns of bakery products—a review. Crit Rev Food
Sci Nutr. 2004;44(1):19‐55.
8. Martucci JF, Gende LB, Neira LM, Ruseckaite RA. Oregano and laven-
der essential oils as antioxidant and antimicrobial additives of biogenic
gelatin films. Ind Crop Prod. 2015;71:205‐213.
9. Neira, L. M. (2017). Películas Comestibles de Gelatina de Pescado con
Actividad Antioxidante. Obtención y Caracterización in vitro e in vivo.
Doctoral Thesis. Universidad Nacional de Mar del Plata.
10. Neira LM, Martucci JF, Stejskal N, Ruseckaite RA. Time‐dependent
evolution of properties of fish gelatin edible films enriched with carva-
crol during storage. Food Hydrocoll. 2019;94:304‐310.
NEIRA ET AL.
11. Kuorwel K, Cran MJ, Sonneveld K, Miltz J, Bigger SW. Essential oils
and their principal constituents as antimicrobial agents for synthetic
packaging films. J Food Sci. 2011;76(9):R164‐R177.
12. Rodriguez‐Lafuente A, Nerin C, Batlle R. Active paraffin‐based paper
packaging for extending the shelf life of cherry tomatoes. J Agric Food
Chem. 2010;58(11):6780‐6786.
13. Dehghani S, Hosseini SV, Regenstein JM. Edible films and coatings in
seafood preservation: a review. Food Chem. 2018;240:505‐513.
14. Ju J, Xie Y, Guo Y, Cheng Y, Qian H, Yao W. Application of edible coat-
ing with essential oil in food preservation. Crit Rev Food Sci Nutr.
2018;1‐14. https://doi.org/10.1080/10408398.2018.1456402
15. Burt S. Essential oils: their antibacterial properties and potential appli-
cations in foods—a review. Int J Food Microbiol. 2004;94(3):223‐253.
16. Mokhtar SM, Youssef KM, Morsy NE. The effects of natural antioxi-
dants on colour, lipid stability and sensory evaluation of fresh beef
patties stored at 4°C. J Agroalim Proces Technol. 2014;20(3):282‐292.
17. Schelegueda LI, Delcarlo SB, Gliemmo MF, Campos CA. Effect of anti-
microbial mixtures and modified atmosphere packaging on the quality
of argentine hake (Merluccius hubbsi) burgers. LWT‐ Food Sci Technol.
2016;68:258‐264.
18. FDA (2017) Essential oils, oleoresins (solvent‐free), and natural extrac-
tives (including distillates) Substances generally recognized as safe, Vol.
21(6), Foods and Drugs Administration. Department of Health and
Human Services.
19. van Beek TA, Joulain D. The essential oil of patchouli, Pogostemon
cablin: a review. Flavour Fragr J. 2018;33(1):6‐51.
20. Debonne E, Van Bockstaele F, De Leyn I, Devlieghere F, Eeckhout M.
Validation of in‐vitro antifungal activity of thyme essential oil on Asper-
gillus niger and Penicillium paneum through application in par‐baked
wheat and sourdough bread. Lebensmittel‐ Wissenschaft & Technologie
LWT. 2018;87:368‐378.
21. Mani Lopez E, Valle Vargas GP, Palou E, López Malo A. Penicillium
expansum inhibition on bread by lemongrass essential oil in vapor
phase. J Food Prot. 2018;81(3):467‐471.
22. Nanasombat S, Piumnoppakun N, Atikanbodee D, Rattanasuwan M.
Combined effect of cinnamon essential oil and water activity on
growth inhibition of Rhizopus stolonifer and Aspergillus flavus and possi-
ble application in extending the shelf life of bread. Water properties in
food. Health, Pharma Biol Syst: ISOPOW. 2010;10:545‐550.
23. Faccin C, Alberti S, Frare L, Vieira LR, Salas‐Mell MDLM, Freitas EMD.
Bread with yerba mate aqueous extract (Ilex paraguariensis A. St.‐hil.).
Am J Food Technol. 2015;10(5):206‐214.
24. Goncalves ND, de Lima Pena F, Sartoratto A, et al. Encapsulated thyme
(Thymus vulgaris) essential oil used as a natural preservative in bakery
product. Food Res Int. 2017;96:154‐160.
25. Pateiro M, Barba FJ, Dominguez R, et al. Essential oils as natural addi-
tives to prevent oxidation reactions in meat and meat products: a
review. Food Res Int. 2018;113:156‐166.
26. Bourtoom T. Review article edible films and coatings: characteristics
and properties. Int Food Res J. 2008;15(3):237‐248.
27. Gennadios A. Protein‐based films and coatings. Boca Ratón. Florida:
CRC Press; 2002.
28. Alparslan Y, Baygar T, Baygar T, Hasanhocaoglu H, Metin C. Effects of
gelatin‐ based edible films enriched with laurel essential oil on the
quality of rainbow trout (Oncorhynchus mykiss) fillets during refriger-
ated storage. Food Technol Biotechnol. 2014;52(3):325‐333.
29. Andevari GT, Rezaei M. Effect of gelatin coating incorporated with cin-
namon oil on the quality of fresh rainbow trout in cold storage. Int J
Food Sci Technol. 2011;46(11):2305‐2311.
9
30. Cha JY, Cho YJ, Kim CJ, Kim CT. Characteristics of gelatin packaging
film incorporated with propolis for food storage. Food Engin Prog.
2007;11:112‐118.
31. Chandra Mohan CS, Rakhavan Kavindapadi R, Radha Krishnan K, et al.
Impact of S. aromaticum and C. cassia incorporated edible films on
shelf life of seer fish (Scomberomorus guttatus) stored at different tem-
perature conditions. J Food Proces Preser. 2016;41(4):1‐11.
32. Echeverría I, López‐Caballero ME, Gómez‐Guillén MC, Mauri AN,
Montero MP. Active nanocomposite films based on soy proteins‐
montmorillonite‐ clove essential oil for the preservation of refrigerated
bluefin tuna (Thunnus thynnus) fillets. Int J Food Microbiol. 2018;
266:142‐149.
33. Gómez‐Estaca J, Montero P, Giménez B, Gómez‐Guillén MC. Effect of
functional edible films and high pressure processing on microbial and
oxidative spoilage in cold‐smoked sardine (Sardina pilchardus). Food
Chem. 2007;105(2):511‐520.
34. Ahmad M, Benjakul S, Sumpavapol P, Nirmal NP. International journal
of food microbiology quality changes of sea bass slices wrapped with
gelatin fi lm incorporated with lemongrass essential oil. Int J Food
Microbiol. 2012;155(3):171‐178.
35. Kaewprachu P, Osako K, Benjakul S, Suthiluk P, Rawdkuen S. Shelf life
extension for Bluefin tuna slices (Thunnus thynnus) wrapped with myo-
fibrillar protein film incorporated with catechin‐Kradon extract. Food
Control. 2017;79:333‐343.
36. Gómez‐Estaca J, López de Lacey A, López‐Caballero ME, Gómez‐
Guillén MC, Montero P. Biodegradable gelatin e chitosan films incorpo-
rated with essential oils as antimicrobial agents for fish preservation.
Food Microbiol. 2010;27(7):889‐896.
37. Wu J, Ge S, Liu H, et al. Properties and antimicrobial activity of silver
carp (Hypophthalmichthys molitrix) skin gelatin‐chitosan films incorpo-
rated with oregano essential oil for fish preservation. Food Pack Shelf
Life. 2014;2(1):7‐16.
38. Kazemi SM, Rezaei M. Antimicrobial effectiveness of gelatin‐alginate
film containing oregano essential oil for fish preservation. J Food Saf.
2015;35(4):482‐490.
39. Schreiber SB, Bozell JJ, Hayes DG, Zivanovic S. Introduction of primary
antioxidant activity to chitosan for application as a multifunctional
food packaging material. Food Hydrocoll. 2013;33(2):207‐214.
40. Aljawish A, Chevalot I, Madad N, Paris C, Muniglia L. Laccase
mediated‐ synthesis of hydroxycinnamoyl‐peptide from ferulic acid
and carnosine. J Biotechnol. 2016;227:83‐93.
41. Fu S, Wu C, Wu T, Yu H, Yang S, Hu Y. Preparation and characteriza-
tion of chlorogenic acid‐gelatin: a type of biologically active film for
coating preservation. Food Chem. 2017;221:657‐663.
42. Curcio M, Puoci F, Iemma F, et al. Covalent insertion of antioxidant
molecules on chitosan by a free radical grafting procedure. J Agric Food
Chem. 2009;57(13):5933‐5938.
43. Liu J, Meng CG, Liu S, Kan J, Jin CH. Preparation and characterization
of protocatechuic acid grafted chitosan films with antioxidant activity.
Food Hydrocoll. 2017;63:457‐466.
44. Spizzirri UG, Iemma F, Puoci F, et al. Synthesis of antioxidant polymers
by grafting of Gallic acid and Catechin on gelatin. Biomacromolecules.
2009;10(7):1923‐1930.
45. Spizzirri UG, Parisi OI, Iemma F, et al. Antioxidant–polysaccharide con-
jugates for food application by eco‐friendly grafting procedure.
Carbohydr Polym. 2010;79(2):333‐340.
46. Guy Carvajal Carranza. Microbiología de Alimentos marinos (I). Lima:
SAGSA, Spencer JFT, Ragout de Spencer AL. Food Microbiology Proto-
cols. Totowa: Humana Press Inc; 2001.
10
47. A.O.A.C, 1990 47 A.O.A.C. Official Methods of Analysis. 15th ed.
Washington, D.C: Association of Official Analytical Chemists; 1990.
48. Giannini DH, Davidovich LA, Lupín HM. Adaptación del método de uso
comercial para la determinación de nitrógeno básico volátil en merluza
(Merluccius hubbsi). Revista de Agroquímica Y Tecnología de Alimentos.
1979;19(1):55‐60.
49. A.O.C.S. (1997). Peroxide Value. In AOCS Official Method Cd 8‐53.
Surplus 2003. Illinois, USA: Official Methods and Recommended Prac-
tices of the American Oil Chemists' Society, Champaign.
50. Bligh EG, Dyer WJ. Canadian journal of biochemistry and physiology.
Can J Biochem Physiol. 1959;37(8):911‐917.
51. A.O.C.S. (1998). p‐Anisidine value. In AOCS Official Method Cd 18‐90.
Illinois, USA: Official Methods and Recommended Practices of the
American Oil Chemists' Society, Champaign.
52. Kamal‐Eldin A, Pokorny J. Analysis of Editors. Illinois, USA: AOCS press;
2005.
53. Pedrero F, Daniel L, Pangborn RM. Evaluación sensorial de los
alimentos. Métodos Analíticos. 1989;641(41):43.
54. Anzaldúa Morales A. La Evaluación Sensorial de los Alimentos en teoría y
la práctica. Zaragoza, España: Acribia; 1994.
55. Dolea D, Rizo A, Fuentes A, Barat JM, Fernández‐Segovia I. Effect of
thyme and oregano essential oils on the shelf life of salmon and sea-
weed burgers. Food Sci Technol Int. 2018;24(5):394‐403.
56. ICMSF (Ed). Sampling plans for fish and shellfish. In: ICMSF (ed.) Micro-
organisms in Foods. Sampling for Microbiological Analysis: Principles and
Scientific Applications. 2nd ed. Vol.2 Toronto: University of Toronto
Press; 1986.
57. FAO. Food and Agriculture Organization. Chemical analysis manual for
food and water. 5th ed. Rome, Italy; 1997.
58. Du WX, Avena‐Bustillos RJ, Woods R, et al. Sensory evaluation of
baked chicken wrapped with antimicrobial apple and tomato edible
films formulated with cinnamaldehyde and carvacrol. J Agric Food
Chem. 2012;60(32):7799‐7804.
59. Friedman M. Chemistry and multibeneficial bioactivities of carvacrol (4‐
Isopropyl‐2‐methylphenol), a component of essential oils produced by
aromatic plants and spices. J Agric Food Chem. 2014;62(31):7652‐7670.
60. Ultee A, Bennik MHJ, Moezelaar R. The phenolic hydroxyl group of
carvacrol is essential for action against the food‐borne pathogen Bacil-
lus cereus. Appl Environ Microbiol. 2002;68(4):1561‐1568.
61. Ibrahim Sallam K. Antimicrobial and antioxidant effects of sodium ace-
tate, sodium lactate, and sodium citrate in refrigerated sliced salmon.
Food Control. 2007;18(5):566‐575.
NEIRA ET AL.
62. Kim JM, Marshall MR, Cornell JA, Preston JF III, Wei CI. Antibacterial
activity of carvacrol, citral and geraniol against Salmonella
Typhimuriumin culture medium\rand on fish Gubes. J Food Sci. 1995;
60(6):1364‐1674.
63. Nilsuwan K, Benjakul S, Prodpran T. Quality changes of shrimp cracker
covered with fish gelatin film without and with palm oil incorporated
during storage. Int Aquatic Res. 2016;8(3):227‐238.
64. Taşkaya L, Çaklı Ş, Kışla D, Kılınç B. Quality changes of fish burger from
rainbow trout during refrigerated storage. EU J Fisheries & Aquatic Sci.
2003;20(1):147‐154.
65. C.A.A. (2017). Código Alimentario Argentino. Capítulo VI. Alimentos
cárneos y afines, 247.
66. Pascual M, Calderón V. Microbiología alimentaria. Madrid, España: S.A.
Díaz de Santos; 2000.
67. 95/149/EC. (1995). Commission Decision of 8 March 1995 fixing the
total volatile basic nitrogen (TVB‐N) limit values for certain
categories of fishery products and specifying the analysis methods
to be used.
68. Giménez B, Gómez‐Guillén MC, Pérez‐Mateos M, Montero P,
Márquez‐Ruiz G. Evaluation of lipid oxidation in horse mackerel patties
covered with borage‐containing film during frozen storage. Food Chem.
2011;124(4):1393‐1403.
69. Rodriguez‐Turienzo L, Cobos A, Moreno V, Caride A, Vieites JM, Diaz
O. Whey protein‐based coatings on frozen Atlantic salmon (Salmo
salar): influence of the plasticiser and the moment of coating on quality
preservation. Food Chem. 2011;128(1):187‐194.
70. Pereira de Abreu DA, Losada PP, Maroto J, Cruz JM. Evaluation of the
effectiveness of a new active packaging film containing natural antiox-
idants (from barley husks) that retard lipid damage in frozen Atlantic
salmon (Salmo salar L.). Food Res Int. 2010;43(5):1277‐1282.
71. Scramlin SM, Newman MC, Cox RB, et al. Effects of oregano oil brine
enhancement on quality attributes of beef longissimus dorsi and
semimembranosus muscles from various age animals. J Food Sci.
2010;75(2):S89‐S94.
How to cite this article: Neira LM, Agustinelli SP, Ruseckaite
RA, Martucci JF. Shelf life extension of refrigerated breaded
hake medallions packed into active edible fish gelatin films.
Packag Technol Sci. 2019;1–10. https://doi.org/10.1002/
pts.2450