Food Hydrocolloids 19 (2005) 951–957

www.elsevier.com/locate/foodhyd

The role of salt washing of fish skins in chemical and rheological

properties of gelatin extracted
B. Giménez*, M.C. Gómez-Guillén, P. Montero
Instituto del Frı́o (CSIC), Ciudad Universitaria, 28040 Madrid, Spain
Received 10 March 2004; revised 23 September 2004; accepted 29 September 2004

Abstract
Dover sole (Solea vulgaris) skins were washed with different salt solutions previous to gelatin extraction by a mild acid pre-treatment. The
effects of NaCl, KCl, MgCl2, MgSO4 and no-salt washing on the mineral content, yield of extraction, molecular weight distribution, gel
strength, aggregation phenomena and viscoelastic properties of gelatin newly dissolved and after overnight cold maturation were evaluated.
Skin washing with NaCl and KCl induced noticeable changes in molecular weight distribution, and consequently in gel strength and
rheological properties, especially when compared with unsalted gelatin preparations. However, salts containing Mg2C gave rise to the
retaining of this bivalent cation in the resultant gelatin, which is detrimental to the gelatin quality.
q 2004 Published by Elsevier Ltd.

Keywords: Fish gelatin; Salt; Washing; Extraction; Gel strength; Viscoelastic properties

1. Introduction improved by using gelatin-modifying materials (Fernán-

Gelatin is a polypeptide obtained by hydrolytic degra-
dation from collagen, the major constituent of animal
connective tissue. It has found widespread use in the
photographic, pharmaceutical and food industries over the
years. In the food industry, gelatin is used as an ingredient to
enhance the elasticity, consistency and stability of food
products, and therefore good rheological properties are
required.
Most commercial gelatins are derived from mammalian
sources, mainly pigskin and cowhide, but for many socio-
cultural reasons alternative sources, like fish skins, are
highly demanded. In addition, it is also interesting the use of
fish skins in the exploitation of by-products from the fish
processing industry. Mammalian and fish gelatins are
different in the gelling and melting temperatures, which
are lower in fish gelatin. Furthermore, the mammalian gels
are also stronger, a characteristic directly related to the
higher hydroxyproline content (Ledward, 1992; Norland,
1990). The rheological properties of fish gelatins could be
* Corresponding author. Tel.: C34 31 5492300; fax: C34 91 5493627.
E-mail address: bgc@if.csic.es (B. Giménez).
0268-005X/$ - see front matter q 2004 Published by Elsevier Ltd.
doi:10.1016/j.foodhyd.2004.09.012
dez-Dı́az, Montero, & Gómez-Guillén, 2001).
One possible means of manipulating the characteristics
of a given gelatin is to trigger interactions by the addition of
salts (Elysée-Collen & Lencki, 1996). As reviewed by
Asghar and Henrickson (1982), electrolytes have a decisive
influence on the biophysical properties (swelling, solubility,
gelatin, viscosity and water-binding capacity) of a protein at
different ionic strengths and pH values (Hermansson, 1975).
Saline ions may either bind directly to the peptide backbone
of collagen, or affect collagen folding indirectly by
interacting with structurally bound-water molecules.
Hydration caused by the disruption of ions of neutral salts
with non-ionic bonds (e.g. hydrogen bonds) of collagen is
described as ‘lyotropic hydration’. Thus, lyotropic agents
may alter water structure around collagen molecules,
interrupt internal hydrogen bonds, or interact with internal
hydrophobic bonds by direct binding at same sites of the
protein chains (Asghar & Henrickson, 1982).
The effect of different salts on the rheological properties
when added to mammalian gelatins is well known
(Harrington & Von Hippel, 1961). However, little infor-
mation is available related to this effect in fish gelatins. In
this sense, Choi and Regenstein (2000) found that NaCl
952 B. Giménez et al. / Food Hydrocolloids 19 (2005) 951–957
depressed the gel strength and melting point when added to
pork and fish commercial gelatins. Sarabia, Gómez-Guillén,
and Montero (2000), stated that the addition of MgSO4,
(NH4)2SO4 or NaH2PO4 at 0.5 M to megrim gelatin was
critical in raising the melting point, whereas chloride salt
acted to reduce it. Fernández-Dı́az et al. (2001), showed that
the addition of MgSO4 at 0.1 and 0.5 M to fish gelatins led to
improve functional properties like melting temperature.
However, the effect of saline solutions (NaCl, KCl, MgCl2
or MgSO4) applied during washing of fish skins, previously
to gelatin extraction, has not been reported. These salts,
especially chlorides, are commonly used to solubilise
myofibrillar proteins, therefore they will help to remove
rests of muscle adhered to the skins, and also may
predispose collagen for a better extraction. In addition,
they may partially be retained in the resulting gelatin, and
consequently may influence its gelling properties.
The aim of this study was to examine rheological
properties (gel strength and viscoelasticity) and chemical
characteristics (mineral content, molecular weight distri-
bution and kinetics of aggregation) of gelatin extracted from
fish skins washed with different saline solutions. For that
purpose, skins from Dover sole (Solea vulgaris) were used,
a species that has been proved to offer a good potential for
gelatin manufacture.
2. Materials and methods
Frozen Dover sole (Solea vulgaris) skins were obtained
from a local market in Madrid, and were stored in the frozen
state until use. All reagents used were of analytical grade.
2.1. Cleaning of dover sole skins
Thawed skins were washed with tap water (1:6 w/v) in a
Stephan homogenizer (Model UM5; Stephan und Söhne
GmbH & Co., Hameln, Germany) at 5 8C for 2 min, and
rinsed with abundant running tap water. Skins were divided
into five sub-batches. Four of the sub-batches were further
washed with different salt solutions again in the Stephan
homogenizer at 5 8C for 2 min, being rinsed with abundant
tap water afterwards. The salt solutions tested were NaCl,
KCl, MgCl2 and MgSO4 (1:6 w/v), at a concentration of
0.8 M. Excess of water was removed by draining the
cleaned skins and squeezing in a manual press.
2.2. Gelatin extraction procedure
Gelatin extraction procedure was carried out according to
Gómez-Guillén and Montero (2001). Briefly, the method
consists in a mild acid swelling step in 0.05 M acetic acid
and subsequent gelatin extraction in distilled water at 45 8C
overnight.
2.3. Mineral anaylisis
Levels of NaC, KC, Mg2C and Ca2C were determined in
gelatin preparations from all sub-batches by atomic
absorption spectrophotometry (Model 5100, Perkin–Ele-
mer, Norwalk, Connecticut, USA). The content of minerals
was expressed by mg/g gelatin.
2.4. Electrophoretic analysis
Gelatin was first dissolved at 4 mg/ml in distilled water at
60 8C and then 3-fold concentrated loading buffer contain-
ing b-mercaptoethanol was added. Protein samples were
heat-denatured 5 min at 90 8C and analysed by SDS-PAGE
according to Laemmli (1970). NaCl, KCl, MgCl2 and
unsalted gelatin preparations were analysed using stacking
and resolving gels at 4 and 5% acrylamide, respectively. In
case of MgSO4 sample, stacking and resolving gels were
prepared at 5 and 10% acrylamide, respectively, to obtain
the protein bands. In all cases, a Mini Protean II unit
(Bio-Rad Laboratories, Hercules, CA) at 25 mA/gel was
used. The loading volume was 15 ml in all lines. Protein
bands were stained with Coomassie brilliant Blue R250.
Type I and type III collagens from fetal calf skin were used
as markers of a-chain, b- and g-component mobilities. A
low-molecular-weight reference kit (Amersham Bios-
ciences) was used as reference for molecular weights of
MgSO4 gelatin preparation, composed of phophorylase b
(97 kDa), albumin (66 kFa), ovalbumin (45 kDa), carbonic
anhydrase (30 kDa), trypsin inhibitor (20 kDa) and a-
lactalbumin (14 kDa).
2.5. Gel strength
The gel strength, according to Gómez-Guillén et al.
(2002), was determined on a 6.67% gel (w/v), formed by
dissolving the dry gelatin in distilled water at 60 8C, and
cooling the solution in a refrigerator at 7 8C (maturation
temperature) for 16–18 h. The gel strength at 7 8C was
determined on a texture analyser (Aname, model XT2i,
Stable Micro Systems, Godalming, England) with a load
cell of 5 kN, cross-head speed 2 mm/s, equipped with a
1.27-cm-diameter flat-faced cylindrical Teflon plunger. The
dimensions of the sample were 3.3 cm diameter and 6 cm
height. Gel strength was expressed as maximum force (in g),
taken when the plunger had penetrated 4 mm into the gelatin
gels, was average of five determinations.
2.6. Viscoelastic properties
Dynamic studies were performed on a Bohlin CRS-10
controlled stress rheometer rotary viscometer (Bohlin
Instrumments Ltd, Gloucestershire, UK) using a cone-
plate geometry (cone angle 48, gapZ0.15 mm). Solutions
were prepared by mechanical stirring for 15–20 min of pre-
weighed (6.67% w/v) dry gelatin in water at 40 8C. Also
 B. Giménez et al. / Food Hydrocolloids 19 (2005) 951–957
the samples were cooled in a refrigerator at 7 8C (maturation
temperature) for 16–18 h overnight to analyse the effect of
maturation time on gelatin. Temperature ramps were
performed at a scan rate of 1 8C/min, frequency 1 Hz, and
oscillating applied stress of 3.0 Pa. The ramps were
implemented from 40 to 6 8C and back to 40 8C for
dissolved gelatin, to study gelatin gelation and subsequent
melting. Gelatin matured overnight (16–18 h) at 7 8C was
also subjected to a temperature ramp from 6 to 40 8C, to
evaluate the effects of maturation time on the gelatin gel.
The melting temperature was taken as the point at which the
phase angle (d) peaks immediately after a sharp increase.
Setting time (gel onset time) was determined as the time in
minutes elapsing between the last temperature of maximum
d and the first temperature of minimum d (gelling point).
The elastic modulus (G 0 ) and viscous modulus (G 00 ) (in Pa)
were taken at 6 8C to compare characteristics at a given
standard temperature. The error in the reproducibility of the
parameters considered in different determinations of a
single sample was 6% or less.
2.7. Kinetics of aggregation at 314 nm
Aggregation of gelatin subunits during the gel formation
can be monitored by changes in absorbance at 314 nm.
Dried gelatin was dissolved in distilled water at 40 8C.
Gelatins from Dover sole skins washed with NaCl, KCl and
without salt were dissolved at 2%, whereas a 0.5% solution
was prepared for gelatins from skins washed with salts of
Mg2C. The increases in absorbance at 314 nm were
measured for 80 min using a spectrophotometer UV-1601
(Model CPS-240, Shimadzu, Japan). During the time of
analysis, the samples were cooled until 7 8C, achieved after
30 min approximately.
3. Results and discussion
3.1. Mineral analysis
Mineral content analysis reveals that the skins from
Dover sole, washed with salts containing Mg2C during the
gelatin extraction process, gave rise to the retaining of this
bivalent cation in the resultant gelatin. As it is shown in
Table 1, these gelatin preparations contained significantly
higher levels of Mg2C than those obtained from skins
washed with NaCl, KCl or washed with any salt addition.
Table 1
Analysis of minerals (mg/g gelatin) in gelatins extracted from Dover sole skins previously washed with different salts (meanGSD)
Ca2C Mg2C
NaCl 0.115G0.004 0.012G0.001
KCl 0.112G0.005 0.010G0.007
MgCl2 0.152G0.001 0.215G0.004
MgSO4 0.180G0.008 0.303G0.005
No salt 0.193G0.017 0.018G0.002
953
On the other hand, Ca2C content is noticeable in gelatins
extracted from skins washed with Mg2C salts and also from
those in which no salts were used to wash. It seems that the
skin washing with NaCl or KCl led to a Ca2C release from
skins in greater proportion, without retention of either NaC
or KC in the resultant gelatin, as it is observed from the
levels of these minerals in the table. Skins washed with
MgSO4 presented the highest level of all the analysed
minerals.
3.2. Gelatin extraction
Gelatin preparations were performed under identical
conditions, with the exception of the washing previous to
acid treatment of the skins. The yield obtained, expressed as
grams of dry gelatin per 100 g of clean skin, was different
depending on the salt used for washing. The skin washing
with KCl and NaCl led to an improvement of the extraction
yield (19 and 16%, respectively), as compared with no salt
washing gelatins (13%). However, the extraction yield was
lower for the skins washed with salts containing Mg2C
(MgCl2, 7.4%; MgSO4 4%). This difference could be
attributed to the bivalent nature of Mg2C, capable to form
ionic bonds between protein collagen chains that difficult
the gelatin extraction.
The solubility in distilled water of the different gelatin
preparations was similar. However, preparations obtained
by skin washing with Mg2C salts showed a higher light
scattering than the others did. The higher turbidity of these
preparations may be due to the solubilisation of material that
shows higher aggregation ability, as well as the presence of
Mg2C cations in the resultant gelatin, which favour these
aggregation phenomena due to the bivalent nature.
3.3. Molecular weight distribution
In order to characterise differences in molecular weight
distribution, gelatin preparations were analysed by poly-
acrylamide gel electrophoresis in the presence of SDS
(Fig. 1). Gelatin extracted with unsalted skin washing
presented a clear different band pattern compared to the rest
of the gelatin preparations. This sample showed a very low
content in intact a-chains and b-polymers. No clear protein
bands were observed, but mainly corresponded to protein
material around and below 100 kDa. Thus, skin washing
without any salt previous to gelatin extraction, gives rise to a
gelatin composed by a considerable amount of low
NaC KC
0.025G0.003 0.041G0.011
0.028G0.002 0.033G0.001
0.075G0.006 0.047G0.001
0.094G0.004 0.075G0.016
0.054G0.007 0.058G0.010
954 B. Giménez et al. / Food Hydrocolloids 19 (2005) 951–957
Fig. 1. Electrophoretic analysis of gelatin preparations (SDS-PAGE) in the presence of 2-b-mercaptoethanol. (a) NaCl, (b) KCl, (c) MgCl2, (d) unsalted and
(e) MgSO4.
molecular weight components. On the contrary, NaCl and
KCl gelatins were characterised by the presence of a
noticeable amount of a-chain polymers, especially b-
components and higher molecular weight polymers. This
molecular weight distribution is consistent with an
increased yield in gelatin extraction as compared to the
gelatin prepared from skins washed with no salt addition.
Skin washing with NaCl or KCl may open the collagen
structure and facilitate the subsequent swelling by inter-
action with acetic acid. As a result, higher molecular weight
polymers could be more easily extracted. MgCl2 gelatin
showed less content of dimmers of a-chains, trimmers and
greater polymers, compared to NaCl and KCl gelatins. As
noted above, the use of Mg2C made extraction more
difficult, rendering a gelatin preparation with lower yield
than with the monovalent cations NaC and KC. MgSO4
gelatin preparation was composed of peptides of very low
molecular weight, and no band pattern was obtained when a
5% acrylamide resolving gel was used. However, when this
sample was analysed in a 10% acrylamide resolving gel,
protein bands were observed corresponding mainly to 20
and 14 kDa peptides. Given that the extraction yield in this
sample was very low (around 4%), this indicates that only
very small collagen fractions could be extracted.
3.4. Viscoelastic properties
A comparison among the viscoelastic properties of the
four gelatine preparations was made after dissolving them at
6.67% (w/v) in distilled water. Fig. 2 shows the elastic (G 0 )
and viscous (G 00 ) moduli, both measured at 6 8C, plotted as a
function of melting temperature, immediately after dissol-
ving (left part of the panel) and after maturing for 16–18 h at
7 8C (right part of the panel).
The MgSO4 gelatin preparation did not show gel forming
capacity. This may be due to the absence of a-chains, along
with the disability of the highly degraded peptides that
compose this gelatin to establish suitable junction zones for
collagen refolding to form a gel. The gelatin without salt
washing presented relatively poor gelling ability, showing
the lowest increase in G 0 and G 00 upon cooling. Furthermore,
MgCl2 sample showed a significant lower increase than
NaCl and KCl samples in both parameters, which presented
quite similar behaviour.
The washing of the skins with NaCl, KCl and MgCl2
previous to gelatin extraction, led to a notable rise of the
thermal stability of the gels, especially in case of NaCl and
KCl. Thus, melting temperature for gelatin preparation with
no salt washing was 7 8C lower than MgCl2, and 9 8C lower
than NaCl and KCl samples. The inferior viscoelastic
properties and melting temperature shown by the gelatin
with no salt washing could be explained by the differences
described in molecular weight distribution. As described by
Gilsenan and Ross-Murphy (2000), low molecular weight
gelatins melt at a lower temperature than high molecular
weight ones, since the lower the molecular weight, the
greater the number of crosslinks per unit volume needed to
form a gel. Moreover, the prevalence of smaller collagen
fractions in the gelatin from skins washed with no salt
probably impairs the formation of suitable strong junctions,
which, as review by Ledward (1992), dictate the melting
point.
The washing with NaCl and KCl gave rise to a prolonged
setting time when compared to the washing with MgCl2
(Table 2; 6 vs. 4 min). The samples without salt washing did
not reach the state of gel in the experimental conditions,
remaining as a viscous sol, as it can be deduced from the
value of phase angle achieved (Table 2). As a consequence
of the predominance of low molecular weight components
in this gelatin preparation, which leads to a limited
availability of junction zones, probably the number of
established cross links would not be enough to form a gel.
As observed for the melting temperature, KCl and NaCl
gelatins showed the highest gelling temperature (13 8C).
The viscoelastic properties of the gelatin preparations
were also analysed after cold maturation at 6 8C for 16–18 h
 B. Giménez et al. / Food Hydrocolloids 19 (2005) 951–957 955

Fig. 2. Elastic modulus (G 0 ) and viscous modulus (G 00 ) measured at 6 8C, plotted as a function of melting temperature, of dissolved gelatins at 6.67% (w/v)
extracted from Dover sole skins previously washed with different salts, upon cooling from 40 to 6 8C and subsequent heating from 6 to 40 8C (left panel) and
after overnight maturation at 7 8C, upon heating from 6 to 40 8C (right panel).

and further heating from 6 to 40 8C (Fig. 2). It is noticeable
the significant increase in elastic and viscous moduli
induced by cold maturation in all the gelatin samples.
After maturation time, NaCl and KCl samples presented the
highest G 0 value (6000 Pa). Although the gelatin without
salt washing exhibited the greater increase in G 0 (2.5-fold
increase), the value (2500 Pa) remained the lowest. During
cold maturation, the limited junction zones of this gelatin
preparation have grown allowing a sufficient collagen chain
annealing to form a gel. NaCl and KCl gelatins showed a
2-fold increase in G 0 , whereas G 0 increased 2.25-fold times
in MgCl2 samples. On the other hand, G 00 increased in all the
gelatin samples as well, especially in gelatin without salt
(6.25-fold increase), and small differences were presented
among gelatins. The thermal stability was also modified by
the cold maturation, increasing in KCl and NaCl samples
from 25 to 27 8C, and from 16 to 20 8C in case of gelatin
without salt washing. These results emphasise the import-
ance of the cold maturation time to promote cross-linking of
collagen chains to form a gel network. In contrast, cold
maturation did not improve melting temperature of MgCl2
gelatin.
It is shown in this study that the skin washing with salts,
especially NaCl and KCl, gives rise to an improvement of
and salt competing for water to hydrate (Elysée-Collen &
Lencki, 1996). It seems that NaCl, when added to a gelatin
solution, is capable of breaking both hydrophobic and
hydrogen bonds, hindering the stabilization of the gel
junction sites (Choi & Regenstein, 2000). However, as a
result of the addition of MgSO4 to a gelatin solution, a rise
in the melting temperature, elastic modulus and gel strength
has been described (Fernández-Dı́az et al., 2001; Sarabia et
al., 2000). This effect has been attributed to the production
of greater screening of electrostatic interactions and
promotion of useful junctions by correct protein unfolding
without distorting the subsequent assembly of chains into
collagen-like helical rods. Nevertheless, when MgSO4 is
used to wash skins previous to gelatin extraction, the gelatin
preparation obtained lacks any viscoelastic behaviour.
3.5. Kinetics of aggregation at 314 nm
The observation of changes in absorbance at 314 nm
makes possible to study associations among denatured
Table 2
Setting time, gelling temperature and phase angle upon cooling from 40 to
6 8C of dissolved gelatins at 6.67% (w/v) extracted from Dover sole skins
previously washed with different salts

the gelatins extracted. In contrast, when NaCl is added to Gelling temperature Setting Phase angle at
the resultant gelatin after extraction, a worsening of the (8C) time (min) 6 8C (degrees)
viscoelastic properties and gel strength is observed (Choi & NaCl 13 6 1.13
Regenstein, 2000; Sarabia et al., 2000). Salts have been KCl 13 6 1.13
MgCl2 11 4 1.10
reported to destabilize gelatin structure (Slade & Levine,
No salt !6 O4 3.45
1987), probably as a direct consequence of both protein
956 B. Giménez et al. / Food Hydrocolloids 19 (2005) 951–957
collagen/gelatin polypeptide chains, which may be directly
responsible for the enhancement of the nucleation step
required for triple helix formation and gelatin gels
stabilisation. Fig. 3 represents the kinetics of aggregation
of the studied gelatins during cooling from 38 to 7 8C. To
avoid artefacts in the interpretation of the kinetics curves, all
the gelatins were diluted to present an initial absorbance
lower than 0.5.
MgCl2 and MgSO4 gelatin preparations (0.5% w/v)
showed a significantly higher absorbance than the rest of
the samples (2% w/v), especially than NaCl and KCl
samples, even though the former presented a lower degree
of covalently linked aggregates. This may be due to the
presence of Mg2C in the resultant gelatins. As bivalent
cation, Mg2C can lead to aggregation of gelatin chains by
forming non-covalent interactions, which are independent
from temperature. Gelatin preparation without any salt also
presented higher non-covalent aggregation among gelatin
chains in solution than NaCl or KCl samples. As derived
from electrophoresis analysis, the gelatin is composed of
very short peptides, which may interact through bivalent
cations such us Ca2C or Mg2C, present in the resultant
gelatin, inducing aggregation. Furthermore, it has been
shown that the smaller the peptide gelatin chains, the higher
the probability of random interactions and more difficult the
folding of collagen upon cooling, since a higher amount of
disorder helix results, far from the original triple-helix
structure of collagen.
When the evolution of the kinetics upon cooling is
compared, the obtained curves are similar to those
described by Djabourov, Lechaire, and Gaill (1993) for
collagen aggregation, with a short lag phase, followed by
a sharp growth step until a plateau is achieved. In the
collagen aggregation kinetic curves, the growth step is
related to microfibrillar assembling by lateral aggregation
Fig. 3. Changes in absorbance at 314 nm upon cooling of gelatin preparations. Dry gelatins were dissolved at 0.5 or 2% (w/v) at 40 8C and loaded into preheated
cuvettes. Changes in the absorbance at 314 nm were monitored during cooling from 40 to 7 8C approximately 1 8C/min and for a total time of 80 min. (6)
Temperature (8C), (,) Unsalted, (:) MgCl2, (B) MgSO4, (&) KCl and (C) NaCl.
of collagen sub-units. In the present study, a random side-
by-side or end-by-end aggregation of gelatin chains may
occur upon cooling, simultaneously to triple helix
development. Such mechanism of aggregation reaches an
equilibrium point in NaCl and KCl preparations, whereas
it persists in time in gelatins without salt and, especially,
in MgCl2 gels. It has been shown that circular dichroism,
as and index of triple helix development, and the
viscoelastic behaviour in gelatins are very related
(Gómez-Guillén et al., 2002). However, it seems that
helical structure formation is independent from this
random aggregation. Thus, it was found that MgSO4
gelatin sample was not able to establish sufficient triple
helix structures to show any viscoelastic behaviour.
Nevertheless, the short peptide chains that compose it
presented a high ability to aggregate due to the SO2K4 and/
or Mg2C content, bivalent ions both of them.
The observed differences may be attributed to the
effect derived from the use of different salts to wash the
fish skins previous to gelatin extraction. The bivalent
nature of Mg2C gives rise to ionic bonding between
collagen chains, hindering the posterior swelling of
collagen chains by acetic acid. As a consequence, both,
the yield of the extraction and the amount of alpha chain
polymers released in the extractive procedure, are low.
Furthermore, as shown before, Mg2C is retained and
present in the resultant gelatin, inducing aggregation and
disturbing the triple helix forming ability. As compared to
gelatin with no salt washing, it seems that the use of NaCl
or KCl to wash the skins before extraction separates the
protein collagen chains. This facilitates the incorporation
of acetic acid and the consequent swelling by the non-
ionised form of the organic acid, which cleaves hydrogen
bonds and reduces the molecular size of the chains. A
reduced molecular size would favour intermolecular
 B. Giménez et al. / Food Hydrocolloids 19 (2005) 951–957
associations and facilitate better stabilization of the
collagen helix, enhancing rheological properties of the
gelatins (Ledward, 1986).
3.6. Gel strength
The gel strength after overnight maturation at 7 8C varied
considerably depending on the salt treatment used for skin
washing. The highest gel strength was achieved in NaCl and
KCl gelatins (181.74G10.19 and 176.83G11.57, respect-
ively). In contrast, a soft gel was obtained when MgCl2 or no
salt were used (82.84G3.80 and 78.24G5.42 g, respect-
ively). These differences could be explained in part by
differences in molecular weight distribution. According to
the electrophoretic gel in Fig. 1, NaCl and KCl gelatins
presented the largest amount of a-chain polymers (dimmers
and trimmers), which allow better stabilising interactions to
form more organised triple helical structures (Sims, Bailey,
& Field, 1997; Stainsby, 1987). However, additional factors
may also influence gel strength as judged from the similar
values offered by the MgCl2 gelatin and the unsalted one,
having a very different average molecular weight. The low
molecular weight gelatin obtained by washing without salt
addition could reasonably explain the decrease in gel
strength. However, in case of MgCl2 gelatin preparation,
which present a significant amount of b-dimmers, the high
content in both, Mg2C and Ca2C, may interfere with gel
strength development.
4. Conclusions
Fish skins washed with NaCl and KCl at 0.8 M led to an
improvement in collagen extraction and its transformation
into soluble gelatin. An adequate amount of a-chain and its
polymers (dimmers and trimmers) was obtained and the
release of Ca2C from skin collagen was favoured, leading to
a gelatin with a low content in saline ions. This resultant
gelatin presented a higher gelling ability and higher stability
(melting and gelling points) than those obtained from skins
without salt washing.
The skins washed with MgCl2 previous to gelatin
extraction impaired the extraction of polymers of a-chains
and gave rise to the presence of a high Mg2C level in the
resultant gelatin. Both characteristics act to the detriment
of gel strength. The MgCl2 gelatin preparations presented
a higher turbidity as a consequence of the existence of
non-covalent interactions among the peptide chains,
which favour the random aggregation side-by-side or
end-to-end, independently of the triple helix forming
ability.
957
Acknowledgements
This research was financed by the Comunidad de Madrid
under project 07G/0052/00.
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