Efecto de la adición de quitosana en la inhibición de la oxidación
lipídica en carne de cerdo molida
J. Food Sci. Gastron
. (January - June 2023) 1(1): 1-7
https://doi.org/10.5281/zenodo.13974644
ISSN: 3073-1283
ORIGINAL ARTICLE
Effect of chitosan addition on the inhibition of lipid
oxidation in ground pork
Mario A. García
mario.garcia@sangregorio.edu.ec
1 Instituto de Farmacia y Alimentos, Universidad de La Habana,
Cuba.
2 Universidad Técnica de Manabí, Ecuador.
3 Centro de Investigación y Desarrollo de Medicamentos,
La Habana, Cuba.
Received: 12 December 2022 / Accepted: 16 January 2023 / Published online: 31 January 2023
© The Author(s) 2023
Jorge Cruz
1
·
Mario A. García
2
·
Nilia de la Paz
3
Abstract
The objective of this study was to evaluate the
efect of UV irradiation on chitosan’s antioxidant activity us
-
ing the ABTS assay and its inhibition of lipid oxidation in
ground pork through the TBA assay. Chitosan was obtained
from lobster chitin (
Panulirus argus
) through a thermo-al
-
kaline N-deacetylation process. Six treatments were applied
at diferent times (5, 15, and 30 min) and two wavelengths
(254 and 365 nm). A sample of non-irradiated chitosan was
used as a comparison standard. The chromatic coordinates of
the chitosan solutions were also determined. UV irradiation
of chitosan did not signifcantly vary (
p
>0.05), neither in the
chromatic coordinates nor in the antioxidant activity of its
1% (w/v) solutions under the tested conditions. Adding the
chitosan solution in 1% (v/v) lactic acid at a ratio of 2.5 mL
per 50 g of ground pork reduced lipid oxidation from 0.23 to
0.14 mg MDA/kg after 24 hours at room temperature.
Keywords
chitosan, irradiation, antioxidant activity, lipid
oxidation, ground pork.
Resumen
El objetivo del presente trabajo fue evaluar el
efecto de la irradiación UV de la quitosana en su actividad
antioxidante mediante el ensayo ABTS e inhibición de la
oxidación lipídica en carne de cerdo molida mediante el
ensayo TBA. Se utilizó quitosana obtenida de la quitina de
langosta (
Panulirus argus
), a través de un proceso de N-des
-
acetilación termo-alcalina. Se aplicaron seis tratamientos a
diferentes tiempos (5; 15 y 30 min) y dos longitudes de onda
(254 y 365 nm). Como patrón de comparación, se empleó
una muestra de quitosana no irradiada. También se deter
-
minaron las coordenadas cromáticas de disoluciones de las
quitosanas. La irradiación UV de la quitosana no varió sig
-
nifcativamente (
p
>0,05), ni las coordenadas cromáticas, ni
la actividad antioxidante de sus disoluciones al 1 % (m/v)
en las condiciones ensayadas. La adición de la disolución de
quitosana en ácido láctico al 1 % (v/v) a razón de 2,5 mL por
50 g de carne de cerdo molida, redujo su oxidación lipídica
desde 0,23 hasta 0,14 mg MDA/kg a las 24 h a temperatura
ambiente.
Palabras clave
quitosana, irradiación, actividad antioxidan
-
te, oxidación lipídica, carne de cerdo molida.
How to cite
Cruz, J., García, M.A., & de la Paz, N. (2023) Efect of chitosan addition on the inhibition of lipid oxidation in ground pork.
Journal of Food Science and
Gastronomy
, 1(1), 1-7. https://doi.org/10.5281/zenodo.13974644
J. Food Sci. Gastron
. (January - June 2023) 1(1): 1-7
2
Introduction
Food safety remains a critical public health issue due to the
constant outbreaks of foodborne diseases. In recent years,
an increase in food poisoning, especially in animal-origin
foods, has been observed. This is attributed, in part, to the
growth of global trade, changes in food production, and new
lifestyles that have altered food consumption patterns.
According to the FAO and WHO, food contamination cau
-
ses signifcant economic losses in the food industry. There
-
fore, various chemical, physical, and new technological pro
-
cesses have been developed to extend the shelf life of foods,
highlighting the use of natural antioxidants as a promising
solution. In this context, chitosan has become as an efective
preservative for animal-origin foods (No et al., 2007; Dutta
et al., 2009). Moreover, its production from seafood industry
waste, such as crustacean exoskeletons, ofers an opportuni
-
ty to utilize these residues, as they are a rich source of chitin.
Chitin is the second most abundant natural polymer after
cellulose and is widely distributed in nature. Its high repleni
-
shment rate in the biosphere makes it an important renewa
-
ble resource (Hernández et al., 2009). The main source of
chitin comes from crustacean exoskeletons, such as lobsters,
which contain high concentrations of this polymer. Chitosan
can be obtained from chitin through a chemical N-deacetyla
-
tion process. Due to its functional and physicochemical pro
-
perties, chitosan has applications in various felds, including
food, medicine, agriculture, cosmetics, and pharmaceuticals.
Although there are methods for obtaining and characterizing
it, its use remains limited by the variability in its chemical
composition, degree of deacetylation, and chain size (Her
-
nández et al., 2009).
The presence of amino groups in the structure of chitosan
gives it great versatility for making modifcations, such as
enzyme immobilization, grafting reactions, and the creation
of cross-linked flms. These modifcations allow the produc
-
tion of materials suitable for applications in biotechnology,
food, and medicine.
One of the key properties of chitosan in the food industry
is its antioxidant capacity, primarily attributed to its efcien
-
cy in chelating metal ions, and preventing lipid oxidation
(Rhazi et al., 2002; Guibal, 2004). However, the exact me
-
chanism of its antioxidant action is still debated.
Previous studies (Youn et al., 2008) have demonstrated
that high-viscosity decolorized chitosan can be obtained
through UV irradiation without bleaching agents, improv
-
ing the product’s sensory characteristics and facilitating its
commercialization. Additionally, UV irradiation treatment
has shown promise as a large-scale chitosan production tech
-
nology, reducing energy consumption by efectively decol
-
orizing chitosan. However, further research is needed on the
efects of UV irradiation dosage on the properties of chitosan
obtained from lobster chitin (
Panulirus argus
). The objective
of this research was to evaluate the efect of UV irradiation
on chitosan and its impact on antioxidant activity and the
inhibition of lipid oxidation in ground pork.
Materials and methods
Chitosan from common lobster (
P. argus
) was used, and
obtained on a pilot scale at the Natural and Synthetic Pro
-
ducts Production Plant of the Center for Drug Research and
Development in Havana, Cuba (de la Paz et al., 2012). Chi
-
tosan is insoluble in water due to its high molecular weight;
in this study, water was used as a dispersing medium at a
ratio of 16 mL/g of chitosan with constant stirring at 120
rpm to ensure uniform treatment of the polymer during its
UV irradiation.
Six treatments (Table 1) were sequentially applied at room
temperature (28 ± 0.5 ºC) with diferent time intervals (5,
15, and 30 min) and wavelengths (254 and 365 nm) using a
low-intensity UV lamp (YL, Mod. WD-9403E, Beijing Liu
-
yi Instrument Factory, China). Immediately after UV irradia
-
tion, the chitosan samples were fltered and dried at 60 ºC for
4 hours. They were then hermetically sealed until used for
evaluating the antioxidant capacity and chromatic coordina
-
tes of their 1% (w/v) solutions.
Table 1.
UV irradiation treatments of chitosan
TreatmentWavelength (nm) Time (min)
1*
--
2
254
5
315
430
5
365
5
615
730
*
Control treatment.
Chitosan solutions at 1.0% (w/v) were prepared in a 1%
(v/v) lactic acid solution with stirring using a magnetic stirrer
for 2 hours. Previously, Tween 80 was added at 0.1% (v/v) to
the 1% (v/v) lactic acid solution. The total antioxidant capa
-
city was evaluated according to the methodology proposed
by Re et al. (1999) and Chien et al. (2007), with some modi
-
fcations. The color determination of the chitosan solutions
was performed using a spectrophotometric method, accor
-
ding to the methodology described by Casariego (2009). A
spectrophotometer (Shimadzu UV-2401PC UV-VIS, Japan)
was used to obtain the transmittance spectrum in the visible
region between 400 and 700 nm.
The meat was taken from the leg and nerve region of the
same animal. All visible fat was then removed. The meat was
J. Food Sci. Gastron
. (January - June 2023) 1(1): 1-7
3
ground using a food processor (Sumeet, Mumbai, India) and
then divided into two groups: a control group and another
group to which the chitosan solution in 1% (v/v) lactic acid
was added at a ratio of 2.5 mL per 50 g of ground meat.
The mixture was thoroughly combined and packed in sealed
plastic tubes.
The antioxidant capacity was determined using the TBA
method, following the methodology proposed by Vyncke
(1975) with modifcations. The results were expressed as
malondialdehyde in mg/kg of sample.
A two-way analysis of variance was performed using the
Statistics software (version 7, 2004, StatSoft. Inc., Tulsa,
USA) and Duncan’s multiple range test to compare diferen
-
ces between the evaluated samples for p ≤ 0.05.
Results and discussion
Table 2 shows the behavior of the antioxidant capacity of
chitosan DFC (1% w/v) with diferent UV treatments. As can
be seen, the wavelength and application time did not infuen
-
ce (
p
>0.05) the antioxidant capacity of the solutions, with
values ranging from 4.93 to 5.48 mg Trolox/100 mL.
Table 2
. Infuence of UV irradiation of chitosan on the an
-
tioxidant capacity of 1% (w/v) solutions
Treatment
Antioxidant capacity
(mg Trolox/100 mL)
14.93 (1.1) a
24.96 (1.0) a
35.01 (0.8) a
45.38 (1.8) a
55.48 (2.1) a
65.09 (1.3) a
75.12 (1.5) a
Mean (Standard deviation); n= 3.
Diferent letters indicate signifcant diferences (
p
≤0.05) according to Dun
-
can’s multiple range test.
UV irradiation can induce the formation of new polar
groups in the chitosan molecule through photooxidation, ge
-
nerating radicals that, in the presence of oxygen, can give
rise to carbonyl, hydroxyl, and hydroperoxide groups (Ra
-
bek, 1995). In particular, OH
●-
radicals derived from irra
-
diated chitosan are highly reactive and can interact with
macromolecules, producing new radicals and macroradicals
(Rabek, 1995). Given that chitosan contains numerous OH
groups, the formation of a large number of OH
●-
radicals and
macroradicals is possible, which could generate cross-linked
structures and afect the polymer’s antioxidant capacity.
It could be expected that UV irradiation increases chito
-
san’s antioxidant capacity due to photochemical changes,
such as polymer chain scission (Sionkowska et al., 2006),
resulting in the cleavage of the acetal bond (C1 and C4),
which would form new active centers capable of reacting
with highly reactive species like free radicals (Figure 1). Ad
-
ditionally, irradiation could cause amide bond cleavage, lea
-
ding to partial deacetylation of the molecule (Rashid et al.,
2012). The degree of deacetylation of chitosan, determined
by FTIR-ATR, slightly decreases after UV treatment (Mucha
& Pawlak, 2002).
Various studies have evaluated the antioxidant activity of
aqueous chitosan solutions, reporting its ability to scavenge
hydroxyl radicals (Xie et al., 2001) and metals (Xue et al.,
1998). The antioxidant activity of chitosan can be explained
through several mechanisms. One of them is its ability to
neutralize free radicals, where nitrogen at C-2 of the poly
-
mer plays a fundamental role. It has been suggested (Xie et
al., 2001) that this ability is related to the reaction of free
radicals with hydrogen ions from ammonium ions (NH3+)
present in stable chitosan molecules (Yen et al., 2008).
Researchers like Chien et al. (2007) examined the antioxi
-
dant activity of three types of chitosan with diferent mole
-
cular weights, reporting that chitosan with 12 kDa had the
highest antioxidant activity, with 2.15 µmol of Trolox equi
-
valents, compared to 1.46 µmol and 0.89 µmol for chitosans
with 95 kDa and 318 kDa, respectively. The antioxidant ca
-
pacity of natural compounds manifests both in the termina
-
tion stage of the free radical formation mechanism and in
their reducing power, which refects the ability to transfer
electrons, being a signifcant indicator of the potential an
-
tioxidant activity of a compound (Meir et al., 1995).
Kim & Thomas (2007) noted that the ability of chitosan to
scavenge free radicals depends on the polymer’s concentra
-
tion and molecular weight. However, Sweetie et al. (2008)
indicated that chitosan has limited antioxidant capacity, ob
-
taining low values in the DPPH assay. Although the nitro
-
gen atom in chitosan has a pair of unshared electrons that
could be donated, in aqueous solutions, the -NH2 groups
are mostly protonated, which hinders this electron donation.
Additionally, chitosan lacks a readily donatable hydrogen
atom, limiting its capacity as an antioxidant (Schreiber et
al., 2013). In contrast, phenolic compounds, classifed as pri
-
mary antioxidants, act by donating a hydrogen atom or an
electron, and the resulting phenoxyl radicals are stabilized
through electron delocalization in the aromatic ring (Eskin
& Przybylski, 2000; Leopoldini et al., 2011).
Generally, a trend has been observed in which the antioxi
-
dant capacity of chitosan increases as its molecular weight
decreases, as reported in previous studies. However, it is
important to consider several factors that can infuence the
results, such as the polymer concentration, the ratios be
-
tween the reagent and the sample, and diferences between
J. Food Sci. Gastron
. (January - June 2023) 1(1): 1-7
4
the chitosans used in each investigation, which limits direct
comparisons of the results.
According to Frankel & Meyer (2000), multiple factors
infuence the efectiveness of antioxidants in heterogeneous
and complex systems such as foods and biological systems.
These factors include the properties of the lipid and aqueous
phases of the antioxidant, the oxidation conditions, and the
physical state of the substrate susceptible to oxidation. Sin
-
ce the infuence of all these parameters cannot be evaluated
with a single test method, the cited studies have employed
diferent techniques to determine the antioxidant capacity of
chitosan. All the studies agree on demonstrating chitosan’s
protective efect against oxidation reactions.
The diferent assays used to estimate antioxidant capacity
allow either for evaluating whether a compound can act as
an antioxidant in one or more ways,
in vivo
or in food matri
-
ces. They can also indicate that antioxidant action is possible
when a compound shows in vitro protection at concentra
-
tions relevant to foods or biological systems. However, an
antioxidant that works in vitro will not necessarily be efecti
-
ve in vivo or in foods, as it might not be absorbed, might not
reach the appropriate site of action, or might be rapidly me
-
tabolized into inactive products (Halliwell, 2002). Therefore,
it is crucial to evaluate the efect of chitosan coatings as an
active packaging method to inhibit lipid oxidation in foods.
Finally, the L*, a*, and b* values of the chitosan solutions
presented in Table 3 show that neither the wavelength nor the
exposure time signifcantly infuenced (
p
>0.05) the chroma
-
tic coordinates of the solutions, with luminosity (L*) values
ranging between 67.39 and 70.04.
When analyzing the values of the component a*, it was
observed that there were no signifcant diferences between
each treatment. The b* value is the parameter that describes
the color of the solutions, due to the yellow color of the chi
-
tosan, and this is the chromatic component that most infuen
-
ces the total color diference (ΔE*) between the chitosans.
The chromaticity values (C*) showed a similar behavior to
that described for the b* component. It is widely accepted
that color changes are related to possible chemical and bio
-
logical changes in a substance. It should be noted that not
only the source of chitosan but also the extraction process
infuences its color (Peniche, 2006).
Youn et al. (2007) reported that the color values for chi
-
tosan dried in the sun for 4 hours were: L* = 86.53; a* =
-0.98; and b* = 10.4, values that are much lower than tho
-
se obtained in this study, which could be due to the factors
infuencing color and may also be afected by the intensity
of UV light used. Among the factors that did not afect the
decolorization are the chitosan/water ratios and the stirring
speed, as neither of these factors signifcantly infuenced co
-
lor; they were set according to the best results obtained from
previous studies (Youn et al., 2008).
Figure 1.
Bond breakage in the chitosan molecule.
Table 3
. Effect of UV irradiation of chitosan on the color of the solutions
TreatmentL*a*b*C*
170.04 (0.372) a1.96 (0.21) a13.11 (0.56) a13.25 (0.42) a
267.83 (0.005) a1.90 (0.04) a12.64 (0.66) a12.76 (0.46) a
368.61 (1.843) a3.10 (1.28) a12.64 (0.64) a13.04 (0.22) a
467.39 (0.013) a1.67 (0.06) a13.19 (0.80) a13.28 (0.55) a
570.01 (0.437) a1.83 (0.04) a13.77 (0.34) a13.89 (0.24) a
669.26 (0.596) a1.62 (0.21) a12.50 (0.05) a12.60 (0.01) a
768.44 (0.647) a1.99 (0.25) a13.27 (0.57) a13.41 (0.42) a
Mean (Standard deviation); n = 2.
Diferent letters indicate signifcant diferences (
p
≤0.05) by Duncan’s multiple range test.
J. Food Sci. Gastron
. (January - June 2023) 1(1): 1-7
5
Figure 2 shows the efect of inhibiting lipid oxidation in
pork with 2 mL of 1% chitosan solution, where a decrease
in lipid oxidation in the chitosan-treated pork was evident 24
hours after the start of the treatment compared to the control
sample (
p
>0.05), which maintained the highest MDA value.
The decrease in peroxide concentration is due to the
biopolymer acting as a barrier against oxygen difusion onto
the meat, which slows the formation of hydroperoxide deri
-
vatives. The macromolecule also mitigates the impact of se
-
condary oxidation; this is due to chitosan, which has amino
groups capable of reacting with malondialdehyde, causing a
reduction in the levels of this aldehyde to below 1 mg MDA/
Kg, considerably delaying the formation of volatile com
-
pounds that may negatively afect sensory properties.
Figure 2.
Inhibition of lipid oxidation by the addition of chi
-
tosan in ground pork at room temperature for 24 hours.
In other studies, Darmadji & Izumimoto (1994) observed
that the addition of 1% w/v chitosan to ground beef signif
-
cantly reduced the thiobarbituric acid (TBA) value compa
-
red to the control sample, demonstrating that the addition
of chitosan decreases lipid oxidation in meat, resulting in a
desirable efect on the stability of the red color of the product
during storage. Lee et al. (2003) observed that pieces of pork
immersed in chitosan solutions with molecular weights of 30
and 120 kDa at 1% (w/v) had a longer shelf life and lower
lipid oxidation.
Other authors have also reported the use of chitosan as
an antioxidant and concluded that this efect improves the
appearance of meat, related to color, and reduces the unplea
-
sant odor generated by the rancidity of the lipids inherent to
the meat (Kanatt et al., 2008; Rao et al., 2005).
Pasanphan et al. (2010) suggested that chitosan began to
be used as a natural antioxidant alternative due to the abi
-
lity of oligomers from chitosan solutions to trap hydroxyl
radicals through ionic reactions with the amino groups in its
chemical structure.
Kanatt et al. (2004) concluded that lamb meat with irradi
-
ated chitosan increased lipid peroxidation. Ahn et al. (1998)
observed that raw pork vacuum-packed with irradiated
chitosan increased lipid inhibition, resulting in an extend
-
ed shelf life. Many authors have studied this phenomenon,
showing diferent results regarding the inhibition time, but
all conclude that chitosan has excellent properties as an in
-
hibitor of lipid oxidation in meats.
Conclusions
UV irradiation of chitosan did not signifcantly vary
(
p
>0.05) the chromatic coordinates or the antioxidant acti
-
vity of its 1% (w/v) solutions under the tested conditions.
The addition of a 1% (v/v) chitosan solution in lactic acid
at a rate of 2.5 mL per 50 g of ground pork reduced lipid
oxidation from 0.23 to 0.14 mg MDA/kg after 24 hours at
room temperature.
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Conficts of interest
The authors declare that they have no conficts of interest.
Author contributions
Jorge Cruz, Mario A. García and Nilia de la Paz: Con
-
ceptualization, data curation, formal analysis, investigation,
methodology, supervision, validation, visualization, drafting
the original manuscript and writing, review, and editing.
Data availability statement
The datasets used and/or analyzed during the current study
are available from the corresponding author on reasonable
request.
Statement on the use of AI
The authors acknowledge the use of generative AI and
AI-assisted technologies to improve the readability and cla
-
rity of the article.
Disclaimer/Editor’s note
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cations are solely those of the individual authors and contri
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tors disclaim any responsibility for any injury to people or
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