Effect of previous chilled storage on rancidity development in frozen horse mackerel (trachurus trachurus)

J Sci Food Agric 82:1764–1771 (online: 2002)
DOI: 10.1002/jsfa.1261

Journal of the Science of Food and Agriculture

Effect of previous chilled storage on rancidity
development in frozen horse mackerel
(Trachurus trachurus)
Santiago P Aubourg,1* Ines Lehmann2 and Jose´ M Gallardo1
1

Instituto de Investigaciones Marinas (CSIC), Eduardo Cabello 6, E-36208 Vigo, Spain
Bundesforschunganstalt fu¨r Fischerei, Institut fu¨r Fischereitechnik und Fischqualita¨t, Palmaille 9, D-22767 Hamburg, Germany

2

Abstract: Rancidity development during frozen storage (À20 °C) of an underutilised medium-fatcontent fish species, horse mackerel (Trachurus trachurus), was studied. Special attention was given
to the effect of previous chilled storage (0, 1, 3 and 5 days) on the quality of the frozen fish. For this,
chemical (free fatty acid and conjugated diene contents; peroxide value, PV; thiobarbituric acid index,
TBA-i; fluorescent compound formation) and sensory (rancid odour and taste) analyses were carried
out. Hydrolytic rancidity showed an increase with frozen storage time; however, no effect of previous
chilling time was observed on the frozen product. Oxidative rancidity measured by chemical (PV,
TBA-i and fluorescence) and sensory (odour and taste) indices increased with frozen storage time and
also with previous chilling time. Satisfactory quality was maintained up to 7 months of frozen storage
of horse mackerel provided that a short chilling time (not longer than 3 days) was employed.
# 2002 Society of Chemical Industry

Keywords: underutilised fish; chilling; frozen storage; chemical and sensory analyses; rancidity; shelf-life

INTRODUCTION

Fish and other marine species give rise to products of
great economic importance in many countries. The
fish industry is actually suffering from dwindling
stocks of traditional species as a result of drastic
changes in their availability. Thus fish technologists
and the fish trade have turned their attention to some
unconventional sources of raw material.1,2 One such
species is horse mackerel (Trachurus trachurus), a
medium-fat-content fish abundant in the Northeast
Atlantic.3,4 Efforts have been made to utilise it in the
manufacture of several smoked,5 canned,6 frozen7,8
and restructured9 fish products.
During processing and storage, fish quality may
decline as a result of several factors. One of the most
important is oxidation of highly unsaturated lipids,10
which is directly related to the production of offflavours and odours.11,12 Frozen storage has been
widely employed to maintain fish properties before
consumption or use in other technological processes.13,14 However, during frozen storage of fish,
lipid hydrolysis and oxidation have been shown to
occur and to influence fish acceptance.15–17
Before the freezing step is accomplished, adequate
storage techniques of fish material should be employed
to reduce post-capture losses. Among the different onboard handling systems that efficiently cool fish, the

most common one employed is chilling.18,19 During
chilled storage of fish, important changes take place in
the lipid fraction, leading to significant losses of
sensory and nutritional values.20,21
The present study is related to the commercialisation of frozen horse mackerel and aims to study the
effect of previous chilled storage on the stability of

horse mackerel lipids during frozen storage. Chemical
and sensory lipid damage indices were studied to
assess rancidity development and, accordingly, to
evaluate the shelf-life of this species under such
conditions.

MATERIALS AND METHODS
Raw fish, sampling and processing

Fresh horse mackerel (T trachurus) were obtained 10 h
after catching in June 2000. The length of the fish was
in the range 18–24 cm; the weight was in the range
250–280 g. Upon arrival in the laboratory the fish were
stored on ice in an isothermal room (0–2 °C). Individual fish (120 pieces) were randomly distributed into
three batches that were studied independently. Horse
mackerel from each batch (40 individual pieces) were
taken for freezing after 0, 1, 3 and 5 days of chilled
storage.
Freezing was carried out at À80 °C for 24 h. After

* Correspondence to: Santiago P Aubourg, Instituto de Investigaciones Marinas (CSIC), Eduardo Cabello 6, E-36208 Vigo, Spain
E-mail:
Contract/grant sponsor: Co-operation Program (1999–2000) Germany–Spain (MCyT–INIA) of Agricultural Research
Contract/grant sponsor: Comisio´n Interministerial de Ciencia y Tecnologı´a; contract/grant number: ALI 99-0869
(Received 31 October 2001; revised version received 14 May 2002; accepted 5 August 2002)

# 2002 Society of Chemical Industry. J Sci Food Agric 0022–5142/2002/$30.00

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Rancidity development in frozen horse mackerel

that time, all fish samples were stored at À20 °C.
Analysis of fish samples was undertaken on the white
muscle of fish material stored for 0, 1, 3, 5 and 7
months at À20 °C.
Composition analyses

Chemicals employed (solvents and reactants) were
reagent grade (E Merck, Darmstadt, Germany).
Water content was determined by weight difference
between the fresh homogenised muscle (1–2 g) and
after heating for 24 h at 105 °C. Results were calculated as g water per 100 g muscle.
Lipids were extracted by the Bligh and Dyer22
method. Results are expressed as g lipid per 100 g wet
muscle.
Chemical lipid damage measurements

Free fatty acid (FFA) content was determined by the
Lowry and Tinsley23 method based on complex
formation with cupric acetate/pyridine. Results are
expressed as g FFA per 100 g lipid. Conjugated diene
(CD) formation was measured at 233 nm.24 Results
are expressed according to the formula CD = BV/w,
where B is the absorbance reading at 233 nm, V is the
volume (ml) and w is the mass (mg) of the lipid extract
measured. The peroxide value (PV), expressed as meq
oxygen kgÀ1 lipid, was determined by the ferric thiocyanate method.25 The thiobarbituric acid index
(TBA-i) was determined according to Vyncke.26

Results are expressed as mg malondialdehyde kgÀ1
fish sample.
Interaction compound formation

Interaction compounds formed between lipid oxidation products and protein-like compounds were
investigated by measuring fluorescence formation
(Perkin-Elmer LS 3B) at 327/415 and 393/463 nm as
described in previous studies.27,28 The relative fluorescence was calculated as RF = F/Fst, where F is the
fluorescence measured at each excitation/emission pair
and Fst is the fluorescence intensity of a quinine
sulphate solution (1 mg ml À1 in 0.05 M H2SO4) at the
corresponding wavelength. The fluorescence ratio
calculated as FR = RF393/463 nm/RF327/415 nm was
studied in the aqueous phase resulting from lipid
extraction.22
Sensory analysis

Sensory analysis was conducted by a trained tasting
panel consisting of six to nine experienced judges. For
each sample analysis, fillets of five fish were cooked in
plastic bags in a water bath. Rancid odour and taste
were then evaluated on a scale from 0 (stage of no
rancidity at all) to 100 (stage where no increase in
rancidity is possible). Three categories were ranked:
good quality (0–29), fair quality (30–59) and rejectable quality (60–100). Scores among panellists were
averaged.
J Sci Food Agric 82:1764–1771 (online: 2002)

Statistical analyses

Data from the different chemical and sensory quality
measurements were subjected to one-way ANOVA
and correlation analysis (p < 0.05);29 comparison of
means was performed using a least squares difference
(LSD) method.

RESULTS AND DISCUSSION

Water content ranged between 750 and 790 g kgÀ1.
Lipid content ranged between 6.0 and 20.0 g kgÀ1.
Differences in both constituents may be explained as a
result of individual fish variation, and not arising from
chilled or frozen storage; it is recommended to employ
several individual fish for each sample to minimise this
source of variation. Comparison of the present results
with previous research showed a higher water content
for horse mackerel than for fattier fish species27 and a
lower water content for horse mackerel than for leaner
fish species,30,31 in accordance with an inverse ratio
between water and lipid matter.32
Lipid hydrolysis

In the present study the FFA content of the raw
material (7.9 g kgÀ1) was similar to that of fatty fish
species (tuna, sardine)27,33 and lower than that of lean
fish species (blue whiting, haddock, cod).30,31 As a
result of frozen storage, significant (p < 0.05) differences were only observed at month 3, where a sharp
increase in the FFA content of all samples was found
(Fig 1). For each frozen storage time, few significant
(p < 0.05) differences could be observed among the

four different chilling times studied, so that an effect of
previous chilling time on hydrolysis during frozen
storage could not be concluded.
Lipid oxidation

Oxidised compounds as measured by CD formation
(Fig 2) did not show a clear trend as a result of frozen
storage, nor as a result of previous chilled storage.
Increases were found at month 5 in samples previously
chilled for 0 and 1 day and also at month 7 in samples
previously chilled for 0 and 3 days. In the present
study, CD detection did not prove to be sensitive for
following changes arising from the chilling and
freezing treatments employed; this index has been
shown to be more accurate in the case of initial
oxidation and model systems.34,35
Peroxide formation (Fig 3) showed a big increase at
month 1 of frozen storage in all samples; thereafter,
few significant increases were observed. A strong effect
of chilled storage time on peroxide formation during
frozen storage was observed. During the whole frozen
storage period, samples previously chilled for 3 and 5
days showed a higher PV than those previously chilled
for 0 and 1 day, indicating a negative effect of the
previous chilling treatment.
Secondary oxidation as measured by TBA-i showed
a gradual general increase during frozen storage for all
samples (Fig 4). A marked effect on carbonyl com1765

SP Aubourg, I Lehmann, JM Gallardo

Figure 1. Free fatty acid (FFA) content in frozen (0,
1, 3, 5 and 7 months) horse mackerel that was
previously chilled (0, 1, 3 and 5 days).

pound formation during frozen storage was observed
for samples previously chilled for 3 and 5 days,
resulting in higher TBA-i values than those for
samples previously chilled for 0 and 1 day.
Interaction compound formation

Few significant differences in fluorescence development (FR) were observed among samples during the
first 3 months of frozen storage (Fig 5); thereafter a
significant (p < 0.05) increase was detected, which was
especially sharp at month 5 in samples previously
chilled for 5 days and at month 7 in samples previously
chilled for 3 and 5 days. Increasing the length of chilled
storage time prior to freezing led to a slight increase in
the formation of fluorescent compounds.

Formation of fluorescent products as a result of
interactions between lipid oxidation compounds and
nucleophilic molecules (proteins, peptides, etc) has
been reported to be dependent on the formation of
lipid oxidation products (peroxides and carbonyls)
and also on temperature.36,37 In the present experiment, fluorescence formation developed faster in the
later stages (5 and 7 months) of frozen storage, in
accordance with increases in PV and TBA-i values
(Figs 3 and 4).38,39

Sensory evaluation40

Rancid odour showed an increase in all samples at
month 3 (Fig 6). Thereafter a significant increase
could only be assessed at month 7 in samples previ-

Figure 2. Conjugated diene (CD) formation in
frozen (0, 1, 3, 5 and 7 months) horse mackerel that
was previously chilled (0, 1, 3 and 5 days).

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J Sci Food Agric 82:1764–1771 (online: 2002)


Rancidity development in frozen horse mackerel

Figure 3. Peroxide (PV) formation in frozen (0,
1, 3, 5 and 7 months) horse mackerel that was
previously chilled (0, 1, 3 and 5 days).

ously chilled for 3 and 5 days, indicating a negative
effect of previous chilling time on rancid odour.
According to the categories assessed by the trained
panel, samples previously chilled for 3 and 5 days were
judged to be of fair quality at month 7 of frozen
storage; however, those previously chilled for 0 and
1 day were judged to be of good quality at month 7.
Rancid taste increased at month 3 of frozen storage
(Fig 7) and continued to increase with frozen storage

time for most samples. However, compared with most
samples, those previously chilled for 5 days scored
higher values. After 7 months of frozen storage,
samples without chilling pre-treatment were found to
be of good quality, those previously chilled for 1 and 3

days were of fair quality and those previously chilled
for 5 days were rejected.
Correlation analyses

The different chemical and sensory analyses were
tested for correlation with the previous chilled storage
time at each frozen storage time (Table 1).
According to the results shown in Figs 1–5, PV and
TBA-i were the only chemical indices that showed
significant (p < 0.05) linear correlation values with the
chilled storage time at all the frozen storage times
tested. PV showed, in all cases, correlation values
higher than 0.84. Sensory analyses (odour and taste)
also showed satisfactory correlations in most cases,

Figure 4. Thiobarbituric acid (TBA-i) values
obtained in frozen (0, 1, 3, 5 and 7 months)
horse mackerel that was previously chilled (0,
1, 3 and 5 days).

J Sci Food Agric 82:1764–1771 (online: 2002)

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SP Aubourg, I Lehmann, JM Gallardo

Figure 5. Fluorescence (RF) formation in
frozen (0, 1, 3, 5 and 7 months) horse
mackerel that was previously chilled (0, 1, 3
and 5 days).

according to Figs 6 and 7. In all cases, exponential and
logarithmic correlations were also tested; however, the
best results were obtained if linear correlations were
used.
Correlation between the most reliable chemical

indices (PV and TBA-i) and the sensory assessments
was also carried out (Table 2). According to Table 1,
satisfactory correlation values were obtained for raw
samples and also in cases of advanced oxidation degree
(months 5 and 7).

Figure 6. Changes in rancid odour value in frozen
(0, 1, 3, 5 and 7 months) horse mackerel that was
previously chilled (0, 1, 3 and 5 days).

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J Sci Food Agric 82:1764–1771 (online: 2002)


Rancidity development in frozen horse mackerel

Figure 7. Changes in rancid taste value in frozen
(0, 1, 3, 5 and 7 months) horse mackerel that was
previously chilled (0, 1, 3 and 5 days).

CONCLUSIONS

In accordance with previous research on lean8,31 and
fatty15,27 fish species, the present experiment showed
important hydrolytic and oxidative rancidity development during frozen storage of horse mackerel. This
lipid damage was assessed satisfactorily by traditional
chemical indices (FFA, PV, TBA-i and FR) and by
sensory analysis (rancid odour and taste). Indeed,
some chemical analyses (PV and TBA-i) showed
satisfactory correlations with sensory values and
proved to be reliable methods for assessing quality
changes (Tables 1 and 2).
Chilled storage time prior to frozen storage did not
provide a significant effect on hydrolytic rancidity

Table 1. Linear correlation values between lipid indices (chemical and
sensory)a and previous chilling time calculated for each frozen storage time

development (FFA formation) in the frozen product.
However, a negative effect on oxidative rancidity,
according to chemical (PV, TBA-i and FR) and
sensory (odour and taste) parameters, was observed
for fish chilled for 3–5 days prior to frozen storage,
leading to a significant quality loss in the frozen
product.

These results concur with previous studies on fattier
species (frozen herring41–43 and canned sardine28,44)
indicating a faster increase in oxidation product
formation when pre-freezing chilled storage was
extended.
In the present study on the medium-fat-content
species horse mackerel, satisfactory quality was maintained up to 7 months of frozen storage provided that
onboard fish handling was not longer than 3 days. In
cases where longer chilling pre-treatments are needed

Frozen storage time (months)
Lipid index
FFA
CD
PV
TBA-i
FR
Odour
Taste

0

1

3

5

7

0.34
À0.76*
0.92*
0.58*
0.60*
0.89*
0.89*

0.37
À0.32
0.85*
0.84*
0.30
0.06
0.77*

0.20
À0.13
0.93*
0.67*
0.27
0.30
0.43

0.33
À0.63*
0.91*
0.75*
0.57
0.72*

0.69*

0.21
À0.28
0.85*
0.77*
0.72*
0.96*
0.96*

a
Abbreviations: FFA, free fatty acids; CD, conjugated dienes; PV, peroxide
value; TBA-i, thiobarbituric acid index; FR, fluorescence ratio.
* Significant at p < 0.05.

J Sci Food Agric 82:1764–1771 (online: 2002)

Table 2. Correlation values between sensory (odour and taste) and chemical
(PV and TBA-i)a indices calculated for each frozen storage time

Frozen storage time
(months)
0
1
3
5
7

Odour (PV/TBA-i)

Taste (PV/TBA-i)

0.87*/0.99*
0.06/À0.03
0.13/0.01
0.78*/0.78*
0.96*/0.92*

0.82*/0.56*
0.61*/0.73*
0.23/0.11
0.77*/0.73*
0.85*/0.90*

a
Abbreviations as specified in Table 1.
* Significant at p < 0.05.

1769


SP Aubourg, I Lehmann, JM Gallardo

for further frozen horse mackerel commercialisation,
employment of chilling or frozen storage in conjunction with appropriate antioxidant treatment45,46 is
recommended to guarantee a longer shelf-life when
consumed as a frozen product.

16

17

ACKNOWLEDGEMENTS

The authors thank Mr Marcos Trigo, Mrs Janet Ares
and Mrs Sybille So¨lter for technical assistance and are
grateful for the financial support provided by the Cooperation Program (1999–2000) Germany–Spain
(MCyT–INIA) of Agricultural Research and the
Comisio´n Interministerial de Ciencia y Tecnologı´a
(project ALI 99-0869; 2000–2002).

18

19

20

21

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