Food Research International 100 (2017) 757–763

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Food Research International

journal homepage: www.elsevier.com/locate/foodres

Application of electrolyzed water for improving pork meat quality MARK

Dirceu Rodrigues Athayde, Diego Rafael Martins Flores, Jéssica Soares da Silva,
Ana Luísa Gomes Genro, Marianna Stafanello Silva, Bruna Klein, Renius Mello,
Paulo Cezar Bastianello Campagnol, Roger Wagner, Cristiano Ragagnin de Menezes,
Juliano Smanioto Barin, Alexandre José Cichoski⁎
Departamento de Tecnologia e Ciência dos Alimentos, Universidade Federal de Santa Maria, Santa Maria 97105-900, RS, Brazil

A R T I C L E I N F O A B S T R A C T

Keywords: The microbiological and oxidative qualities of pork loin sprayed with different types (slightly acidic, acidic and
Acidic electrolyzed water basic) and combinations of electrolyzed water (EWs) were evaluated. The EWs were applied at two temperatures
Basic electrolyzed water (18° and 30 °C) and pressures (30 and 45 psi) and the volume corresponded to approximately 10% water
Slightly acidic electrolyzed water commonly used in carcass washing. EW after spraying exhibited a chlorine concentration between 15 and
Spray
25 ppm. The application of acidic EW (AEW) alone or in combination with basic EW (BEW) decreased
(P < 0.05) the microbial counts shortly after spraying. In addition, the combination of BEW + AEW (30 psi)
reduced the mesophilic and psychrotrophic bacteria counts throughout the refrigerated storage (P < 0.05). The
EWs did not increase the lipid oxidation of the samples. On the other hand, a high protein oxidation was ob-
served in the samples sprayed with AEW and slightly acidic EW (SAEW), while BEW was effective to reduce the
oxidation reactions. Therefore, the results showed that the combination BEW + AEW may be a viable alternative
to reduce the volume of water used at slaughter and to improve the microbiological quality of pork meat.

1. Introduction
Washing of pork carcasses after toileting prior to refrigerated sto-
rage is performed in most industries, using water at room temperature
and pressure of 3 atm, and a large amount of water per carcass (Cattani,
2012). Electrolyzed water (EW) is an emerging technology with po-
tential for industrial application because it returns to its original state
after application since chlorine species are consumed during contact
with organic matter (Al-Haq, Sugiyama, and Isobe, 2005). The acidic
EW (AEW) has pH between 2 and 3 and oxi-reduction potential
(ORP) > 1000 mV, whereas the slightly acidic electrolyzed water
(SAEW) has pH between 5 and 7 and ORP of 800–900 mV, and the basic
EW (BEW) has pH 10–12 and ORP < − 600 mV. Various chlorine
concentrations (CLC) can be found in different types of EW, as it is
influenced by the brine concentration, electrolysis time, and electric
current applied (Hsu, 2005). The main form of chlorine in AEW is Cl2,
while the hypochlorous acid (HOCl) and hypochlorite (OCl) are most
present in SAEW and BEW, respectively (Rahman, Ding, and Oh, 2010).
The effect of EW on the reduction of microbial counts in several
foods has been widely studied (Cao, Zhu, Shi, Wang, and Li, 2009;
Jadeja & Hung, 2014), which is more effective when compared to so-
dium hypochlorite and peroxyacetic acid (Cao et al., 2009; Mártinez-
Hernández et al., 2015). However, most of the studies evaluated the use
of one type of EW alone as a way to reduce microbial counts in foods
(Rahman, Park, Song, Al-Harbi, and Oh, 2012), and the combination of
different types of EW has not been studied. In addition, there is little
information in the literature about the effect of EW on lipid and protein
oxidation of meat. On the other hand, immersion is the main form of
application of EW (Fabrizio & Cutter 2005, Cao et al., 2009; Xu et al.,
2014). This approach, although effective in reducing microbial counts,
has the disadvantage of using large volumes of EW, which makes its
industrial application impracticable in most cases. Thus, studies that
aim to reduce the volume of EW and increase its antimicrobial effi-
ciency are necessary to enable the application of this emerging tech-
nology on an industrial scale.
Based on this, different types and combinations of EW were sprayed
on pork loins. The microbiological quality of the pork loins and the lipid
and protein oxidation were evaluated during the refrigerated storage.
2. Materials and methods
2.1. Production of different types of EW
Different types of EW (AEW, SAEW, and BEW) were obtained from a

⁎
Corresponding author.
E-mail address: cijoale@gmail.com (A.J. Cichoski).

http://dx.doi.org/10.1016/j.foodres.2017.08.009
Received 29 June 2017; Received in revised form 1 August 2017; Accepted 2 August 2017
Available online 03 August 2017
0963-9969/ © 2017 Elsevier Ltd. All rights reserved.
D.R. Athayde et al. Food Research International 100 (2017) 757–763

0.05% sodium chloride (Dinâmica, Brazil) using a portable electrolyser
(Envirolyte, Estonia). The pH and ORP were measured by direct im-
mersion of EW using a glass electrode (Digimed, Brazil) and a platinum
electrode (Digimed, Brazil), and the CLC was evaluated according to
APHA (1998) methodology.
2.2. Sampling and processing
Pork loins at 38 °C ± 1 were collected on 3 different days in a
slaughter in the central state of Rio Grande Sul, Brazil. The pork loins
were cut in 6 units for each application (250 ± 50 g, containing skin,
fat, and meat), and subjected to EW spraying. The temperature of each
loin was 36 °C ± 1 at the time of application, thus simulating the
washing step of the pork carcass after toileting prior to refrigerated
storage. In all experiments, a 10 cm distance was used between the
spray application and the loin, which was rotated at a 360° angle during
application. The pressures of 30 and 45 psi were selected in previous
trials. Four experiments were carried out, as follows: E1: application of
AEW, SAEW, and BEW alone, at 18 °C and 30 psi for 40 s; E2: appli-
cation of AEW, SAEW, and BEW alone at 30 °C and 30 psi for 40 s; E3:
application of BEW for 20 s + AEW for 20 s (BEW + AEW) or appli-
cation of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and
30 psi; and E4: application of BEW for 20 s + AEW for 20 s (BEW
+ AEW) or application of BEW for 20 s + SAEW for 20 s (BEW
+ SAEW) at 18 °C and 45 psi. Each 20 s of spraying corresponded to
100 mL of EW, and a control using deionized water under similar time,
temperature, and pressure conditions was performed. After application,
the pork loins were vacuum packed in low-density polyethylene bags
(24 μ thickness and TPVO2 < 30 cm3/m2·day), and stored at 4 °C ± 1
for 29 days. The determinations were performed at day 1, 15, and 29 of
refrigerated storage (4 °C ± 1) using 2 units for each sampling day.
were expressed as nano mol DNPH/mg protein.
2.7. Thiol groups
The thiol groups were evaluated according to Ellman (1959) with
modifications. For quantification, the absorbance readings were com-
pared to a cysteine standard curve. The absorbance was measured at
412 nm using an Agilent 8453 spectrophotometer (Agilent Co., Ger-
many) after reaction with 5,5′-Dithiobis 2-nitrobenzoic acid (Sig-
ma–Aldrich, St. Louis, MO). The results were expressed as thiol nmol/
mg protein.
2.8. Color measurements
The meat was removed from the vacuum package at 20 °C ( ± 1).
After 30 min, the color measurements were performed using a colori-
meter Minolta CR400 (Konica Minolta, Japan) by direct readings of 5
random points. The illuminant A, specular component included, and
10° observer angle were used to measure the color parameters L*, a*,
and b * (Brewer, Zhu, Bidner, Meisinger, and McKeith, 2001), and the
red index was calculated according to a*/b* (Chen, Chiu, and Huang,
1997).
2.9. Statistical analysis
Data were analyzed by ANOVA, and the differences were analyzed
using the Duncan test (P < 0.05) for comparison between means. The
entire experiment was repeated three times in different days, and each
analysis was carried out in triplicate, using the software STATISTICA
7.0 (StatSoft Inc., USA). The principal component analysis (PCA) charts
were made with SAS.

2.3. Microbiological determinations

3. Results and discussion

Mesophilic (MA) and psychrotrophs (PS) aerobic microorganisms

3.1. Characteristics of different types of EW before and after spraying
were quantified using plate count agar medium (PCA, Merck,
Darmstadt, Germany), lactic acid bacteria (LAB) were quantified using
The concentration of chlorine (CLC) decreasead up to 70% after
Man–Rogosa–Sharpe agar (MRS, Merck, Darmstadt, Germany) by plate
spray due the interaction between chlorine and organic matter.
count technique, with incubation at 36 °C for 48 h for MA and LAB, and
However, a greater reduction of CLC was observed in the EWs at 30 °C
7 °C for 7 days for PS (APHA, 2001).
(AEW-77%, and SAEW-60%) drained from the pork loin, when com-
pared to EWs applied at 18 °C (AEW-69%, and SAEW-51%, Table 1),
2.4. pH and redox potential (ORP)
once that higher temperatures favored the evaporation of Cl2. Probably,
the presence of chlorine in the AEW (23 ppm in E1, E3, and E4; and
For determination of pH and ORP, 5 g sample were homogenized
with 50 mL distilled water with subsequent readings in a pH meter
Table 1
(Digimed, Brazil) according to AOAC (2006). Characteristics of the different types of electrolyzed water used to spray the pork meat.

2.5. Tiobarbituric acid reactive substances (TBARS) Experiments pH POR (mV) Chlorine Post-spray % post-spray
before Chlorine chlorine
spray (mg/L de reduction
TBARS were determined as described by Raharjo, Sofos, and (mg/L de Cl2) (residual watera)
Schimidt (1992), by the reaction of 10 g sample and thiobarbituric acid Cl2)
(Merck, Darmstadt, Germany) at 95 °C for 5 min, followed by absor-
bance readings at 531 nm. The results were expressed as mg of mal- E1 40″ 18 °C
AEW 2.60 1185 74 23 69
onaldehyde (MDA)/kg sample. SAEW 6.5 940 47 23 51
BEW 11.45 − 826 0 0 0
2.6. Carbonyl groups E2 40″ 30 °C
AEW 2.60 1134 74 17 77
SAEW 6.41 875 38 15 60
The carbonylated proteins were determined according to Levine
BEW 11.40 − 877 0 0 0
et al. (1990) with modifications. The protein pellets were dissolved in E3 e E4
2% sodium dodecyl sulfate (SDS, Sigma–Aldrich, St. Louis, MO). The AEW 2.60 1200 74 23 68
samples derived from the precipitate treated with 2,4-dini- SAEW 6.15 930 51 25 51
trophenylhydrazine (DNPH, Sigma–Aldrich, St. Louis, MO) were mea- BEW 11.40 − 830 0 0 0

sured at 370 nm, and a blank was measured at 370 nm for protein
Note: Results expressed as mean.
quantification. The carbonyl group was quantified as [Abs370 with AEW: acidic electrolyzed water; SAEW: slightly acidic electrolyzed water; BEW: basic
DNPH - Abs370 blank] to eliminate the effect of protein absorbance, electrolyzed water.
using the molar extinction coefficient 22,000 M− 1 cm− 1. The results a
Residual water is the water that drained from the pork loin when applied.

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Fig. 1. Relationship between pH and lactic acid bacteria (LAB) on pork loins submitted to different types of electrolyzed water and pressure of spray applications. (E1, E2, E3 and E4) in
vacuum package and refrigerated to 4 °C ± 1 along storage time. E1: application of AEW, SAEW, and BEW alone, at 18 °C and 30 psi for 40 s; E2: application of AEW, SAEW, and BEW
alone at 30 °C and 30 psi for 40 s; E3: application of BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 30 psi; and
E4: application of BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 45 psi. Each 20 s of spraying corresponded to
100 mL of EW, and a control using deionized water under similar time, temperature, and pressure conditions was performed.

17 ppm in E2) and SAEW (23 ppm in E1, 15 ppm in E2 and E3; and
25 ppm in E4) drained from the pork loin could help to sanitize the
surfaces in contact with meat, since studies have shown that the use of
EW with similar chlorine concentration reduced the Listeria mono-
cytogenes biofilms from stainless steel surfaces (Arévalos-Sanchez et al.,
2013), and inhibited the growth of E. coli O157: H7 (Gómez-López,
Lannoo, Gil, and Allende, 2014).
3.2. Microbiological determinations and pH measurements
A significant (P < 0.05) reduction of LAB was observed for both
AEW (E1 and E2) and the combination BEW + AEW (E3 and E4) only
on the 1st day of storage when compared to the control (Fig. 1). A
significant increase was observed in LAB counts (P < 0.05) for all
experiments during the refrigerated storage. However, no differences
were observed between the control and the EW treatments. During the
storage, its normal the pH values of meat increase due to proteolytic
reactions that produce alkaline compounds (Bechet, Tassa, Taillandier,
Combaret, and Attaix, 2005). In our study, the pH practically not in-
creased during the storage. This fact could be attributed to production
of acidic compounds by LAB that practically stabilized the pH values
(Pothakos, Devlieghere, Villani, Björkroth, and Ercolini, 2015). All
types of EW and the different conditions of the present study did not
affect (P > 0.05) the pH of meat after spraying and during storage in
relation to the control (Fig. 1). This suggests that the diluted HOCl and
HCl in SAEW and AEW, respectively, and the diluted hypochlorite and
NaOH in BEW (Rahman et al., 2010) acted only on the bacterial cells,
with no changes in the physicochemical properties of meat. In addition,
the greatest bactericidal action of these electrolyzed waters may have
occurred at the moment of application (spray or immersion).
In the experiments E1 and E2, at day 1, the application of AEW
alone allowed a reduction (P < 0.05) of mesophilic bacteria counts
shortly after spraying, when compared to the control. However, the
application of SAEW and BEW alone was not effective to reduce
(P > 0.05) these microorganisms (Table 2). These results are similar to
those reported by Feliciano, Lee, and Pascall (2012), who found that
AEW (pH 2.7 and 150 mg/L Cl2) was more effective in reducing Listeria
innocua when compared to both SAEW (pH 6.9 and 150 mg/L Cl2) and
BEW (pH 11.6). These authors reported that the EW provided mod-
ifications in the cell membrane and cytoplasm of this microorganism as
observed by transmission microscopy. The combined application of
BEW + AEW or SAEW was effective to reduce the initial mesophilic
bacteria counts (P < 0.05) in relation to the control. In addition, lower
(P < 0.05) mesophilic bacteria counts were observed at the end of
storage (29 days) in the samples subjected to the combined application
of BEW + AEW and BEW + SAEW at 30 and 45 psi, respectively, when
compared to the control.
The psychrotrophic bacteria counts were not affected by application
of AEW, SAEW, and BEW alone at 18 °C during refrigerated storage. On
the other hand, the application of AEW and BEW alone at 30 °C reduced
the initial counts (day 1) by 0.67 and 0.35 Log CFU·g− 1, respectively, in
relation to the control, with no significant differences (P > 0.05) in
other days (15 and 29). Similar to observed in mesophiles counts, the
combination of BEW + AEW and SAEW at 30 psi also reduced
(P < 0.05) the initial psychrotrophic bacteria counts, and the combi-
nation BEW + AEW was also effective to maintain a lower (P < 0.05)
psychrotrophic bacteria counts until the end of storage (day 29). This
higher antimicrobial action can be due the saponification of the fatty
acids in meat surface by the NaOH from BEW, which can facilitate the
action of AEW or SAEW against bacteria (Sun, Zhang, Chen, and Han,

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Table 2
Mesophilic and psychrotrophs counts (Log CFU·g− 1) on pork loins submitted to different types of electrolyzed water and pressures of sprays during refrigerated storage (4 °C ± 1).

Mesophilic microorganisms (Days) Psychrotrophs microorganisms (Days)

1 15 29 1 15 29

E1 AEW 3.67bB (0.28) 6.82aA (0.16) 6.86aA (0.52) 3.41aB (0.53) 6.79aA (0.27) 7.16aA (0.32)
SAEW 4.21abB (1.04) 7.00aA (0.18) 7.39aA (0.47) 3.63aB (1.05) 6.77aA (0.32) 7.45aA (0.12)
BEW 4.67abB (0.21) 7.00aA (0.48) 7.18aA (0.12) 3.92aB (0.44) 7.20aA (0.23) 7.30aA (0.40)
Control 5.34aB (0.08) 7.30aA (0.30) 7.19aA (0.15) 4.59aB (0.26) 7.34aA (0.28) 7.61aA (0.21)
E2 AEW 2.75bC (0.02) 6.80aB (0.30) 7.60aA (0.19) < 10cC1 6.39bB (0.26) 7.13aA (0.29)
SAEW 3.41aB (0.22) 7.03aA (0.34) 7.45aA (0.13) 2.68aC (0.20) 6.93aB (0.19) 7.29aA (0.09)
BEW 3.07abC (0.11) 6.56aB (0.04) 7.54aA (0.14) 1.79cC (0.17) 6.62abB (0.18) 7.23aA (0.14)
Control 3.42aB (0.24) 6.68aA (0.42) 7.20aA (0.49) 2.28bC (0.27) 6.72abB (0.14) 7.25aA (0.28)
E3 BEW + AEW 3.38cB (0.11) 6.71aA (0.31) 7.03bA (0.32) 3.10bB (0.16) 7.13aA (0.31) 6.84bA (0.15)
BEW + SAEW 3.81bC (0.11) 6.69aB (0.27) 7.29abA (013) 3.27bB (0.06) 7.41aA (0.12) 7.31aA (0.27)
Control 4.12aB (0.07) 7.02aA (0.56) 7.49aA (0.10) 3.69aB (0.28) 7.29aA (0.51) 7.66aA (0.13)
E4 BEW + AEW 1.89bC (0.17) 6.55aB (0.12) 7.35aA (0.19) < 101 4.99aB (0.43) 6.57aA (0.08)
BEW + SAEW 2.15bC (0.21) 5.58bB (0.57) 6.79bA (0.25) < 101 5.23aB (0.33) 6.57aA (0.28)
Control 2.92aC (0.25) 6.79aB (0.27) 7.46aA (0.07) < 101 5.61aB (0.53) 6.90aA (0.15)

Note: Lowercase/uppercase different letters on column/lines indicates statistical differences (P < 0.05) within the same experiment by Duncan's test. Results in mean (standard
deviation): E1: application of AEW, SAEW, and BEW alone at 18 °C and 30 psi for 40 s; E2: application of AEW, SAEW, and BEW alone at 30 °C and 30 psi for 40 s; E3: application of BEW
for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 30 psi; and E4: application of BEW for 20 s + AEW for 20 s (BEW
+ AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 45 psi. Each 20 s of spraying corresponded to 100 mL of EW, and a control using deionized water
under similar time, temperature, and pressure conditions was performed.

2012).
3.3. Carbonyl and thiol groups
The carbonyl groups can act as a marker of protein oxidation
(Estévez, 2011), which increases with the increase in oxidation. In this
study, the application of AEW and SAEW alone led to an increase in
carbonyl groups, mainly shortly after application (E1 and E2, Fig. 2).
This result can be due to the molecular species (HCl, Cl2, HOCl) in these
types of EW, which favor the production of carbonylated proteins
(Stadtman & Levine, 2003). On the other hand, the application of BEW
alone or in combination with AEW and SAEW at 30 psi (E3) and 45 psi
(E4) did not lead to an increase in the carbonyl values during storage
when compared to the control (P > 0.05) (Fig. 2). This fact suggests
that the BEW exerted an antioxidant effect, which protected the meat
protein from the AEW and SAEW attack. This result is in agreement

Fig. 2. Relationship between thiol and carbonyl on pork loins submitted to different types of electrolyzed water and pressure of spray applications (E1, E2, E3 and E4) in vacuum package
and refrigerated to 4 °C ± 1 along storage time. E1: application of AEW, SAEW, and BEW alone, at 18 °C and 30 psi for 40 s; E2: application of AEW, SAEW, and BEW alone at 30 °C and
30 psi for 40 s; E3: application of BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 30 psi; and E4: application of
BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 45 psi. Each 20 s of spraying corresponded to 100 mL of EW,
and a control using deionized water under similar time, temperature, and pressure conditions was performed.

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with Lee, Kim, Ryoo, Lee, and Park (2006), who reported the anti- Table 3
oxidant effect of BEW on different types of proteins. TBARS values (MDA/sample Kg) on pork loins submitted to different types of electrolyzed
water and pressures of sprays during refrigerated storage (4 °C ± 1).
Sulfur is present in the amino acids methionine and cysteine and is a
thiol grouping agent. It is very reactive towards oxidizing substances Days
(Birben, Sahiner, Sackesen, Erzurum, and Kalayci, 2012), such as those
present in AEW (HCl, Cl2) and SAEW (HOCl) (Hawkins, Pattison, and 1 15 29
Davies, 2003). Kerkaert et al. (2011) studied the effect of HOCl on bA bA
E1 AEW 0.235 (0.061) 0.173 (0.081) 0.385aA (0.130)
proteins such as casein and found that cysteine, methionine, and tryp- SAEW 0.165bB (0.092) 0.202bAB (0.040) 0.315aA (0.017)
tophan were the most susceptible amino acids to oxidation, and HOCl BEW 0.331abA (0.125) 0.202bA (0.040) 0.312aA (0.057)
led to protein aggregation by forming disulfide bonds (− SS-) between Control 0.424aA (0.029) 0.404aA (0.000) 0.385aA (0.021)
amino acids along the protein chain, which may also lead to a decrease E2 AEW 0.178aA (0.016) 0.063aB (0.032) 0.265cA (0.066)
SAEW 0.226aAB (0.032) 0.151aB (0.150) 0.345bcA (0.037)
in the thiol grouping. In all experiments (E1, E2, E3, and E4) and in all
BEW 0.226aB (0.008) 0.102aB (0.064) 0.438abA (0.108)
periods, the thiol values did not differ (P > 0.05) from the control, Control 0.257aB (0.098) 0.110aB (0.076) 0.504aA (0.036)
demonstrating no oxidative effect of HCl, Cl2, and HOCl present in the E3 BEW + AEW 0.304aA (0.093) 0.346aA (0.152) 0.096aB (0.021)
different types of EW used (Fig. 2). BEW + SAEW 0.312aA (0.109) 0.270aA (0.034) 0.058aB (0.049)
Control 0.234aAB (0.111) 0.347aA (0.115) 0.119aB (0.077)
E4 BEW + AEW 0.238bA (0.055) 0.243bA (0.030) 0.294aA (0.174)
BEW + SAEW 0.322bB (0.064) 0.441aA (0.022) 0.131aC (0.012)
3.4. ORP and color measurements
Control 0.602aA (0.149) 0.296bB (0.038) 0.252aB (0.074)

The ORP value indicates the final concentration of oxidizing and
reducing species in the medium. No differences (P > 0.05) were ob-
served between treatments in the experiments E1, E3, and E4 on the
first day of storage, which remained until day 29 (Fig. 3). In E2, ORP
values in the treatments with the different EW were significantly dif-
ferent from the control at day 15 (P < 0.05), which can be considered
a random effect. The results suggest that the application of EWs did not
affect the ORP of meat (Fig. 3), similar to that occurring with pH values
just after spraying (Fig. 1).
The ORP values decreased (P < 0.05) during storage in the ex-
periments E2, E3, and E4, and increased (P < 0.05) in E1. High ORP
values indicate an environment conducive to oxidation (Jay, Loessner,
and Golden, 2005). In this sense, in E1, the treatment BEW showed the
Note: Lowercase/uppercase different letters on column/lines indicates statistical differ-
ences (P < 0.05) within the same experiment by Duncan's test. Results in mean (stan-
dard deviation): E1: application of AEW, SAEW, and BEW alone at 18 °C and 30 psi for
40 s; E2: application of AEW, SAEW, and BEW alone at 30 °C and 30 psi for 40 s; E3:
application of BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for
20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 30 psi; and E4: application of BEW for
20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s
(BEW + SAEW) at 18 °C and 45 psi. Each 20 s of spraying corresponded to 100 mL of EW,
and a control using deionized water under similar time, temperature, and pressure con-
ditions was performed.
lowest carbonyl values (Fig. 2), and the highest thiols values (Fig. 2),
indicating an environment with little protein oxidation. The ORP values
may be related to carbonyl groups since ORP decreased with an

Fig. 3. Relationship between oxidation-reduction potential (ORP) and index a*/b* on pork loins submitted to different types of electrolyzed water and pressure of spray applications (E1,
E2, E3 and E4) in vacuum package and refrigerated to 4 °C ± 1 along storage time. E1: application of AEW, SAEW, and BEW alone, at 18 °C and 30 psi for 40 s; E2: application of AEW,
SAEW, and BEW alone at 30 °C and 30 psi for 40 s; E3: application of BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at
18 °C and 30 psi; and E4: application of BEW for 20 s + AEW for 20 s (BEW + AEW) or application of BEW for 20 s + SAEW for 20 s (BEW + SAEW) at 18 °C and 45 psi. Each 20 s of
spraying corresponded to 100 mL of EW, and a control using deionized water under similar time, temperature, and pressure conditions was performed.

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increase in the carbonyl values (Fig. 3).
The application of the different types and combinations of EWs did
not affect the red color (a*) of the loins (data not shown), indicating
that the myoglobin pigment did not undergo major changes. The b*
values appeared to be interrelated with the oxidation event, especially
protein, which is observed over time (Chaijan, Benjakul, Visessanguan,
and Faustman, 2005; Li et al., 2011). Thus, the decrease in a*/b* index
with the storage time (Fig. 3) was not significant (P > 0.05) in the
loins sprayed with the different types and combinations of EWs.
3.5. TBARS
The results of TBARS during storage suggested that the application
of AEW, SAEW, and BEW did not accelerate the lipid oxidation in pork
loins (Table 3). In some cases, even the application of AEW and SAEW
alone (E1, Table 3) resulted in lower TBARS values when compared to
the control (P < 0.05), since the malonaldehyde formed by the lipid
oxidation process is susceptible to the combination with other mole-
cules containing amine and thiol groups (Esterbauer, Schaur, and
Zollner, 1991), which may have occurred in these treatments.
3.6. Principal component analysis (PCA) of the experiments
PCA was performed to understand the grouping of samples and the
correlation between the variables. In general, for all experiments, in the
first component 1 (PC1), the samples were plotted as a function of the
storage time, with a high correlation with microbial growth, and higher
weights for the variables mesophiles, psychrotrophs, and lactic acid
bacteria (Fig. 4A, B, C, and D).
In the experiment E1, in addition to the correlation between mi-
crobial growth and storage time, an increase in redox potential (OPR)
and pH of the samples (Fig. 4A) was observed, as shown in PC1.
Fig. 4. Canonical discriminant analysis to steps of experiments (E1, E2, E3 and E4).
762
Food Research International 100 (2017) 757–763
However, PC2 discriminated treatment groups from control at 18 °C,
mainly at days 1 and 15. In this component, the variables TBARs and
thiol groups were correlated with BEW and control, while the carbonyl
groups were more correlated with AEW and SAEW. These results are
probably due to the antioxidant effect of BEW on proteins (Lee et al.,
2006), and the oxidizing action of the molecular species (HCl, Cl2,
HOCl) in AEW and SAEW, which can favor the production of carbo-
nylated proteins (Stadtman & Levine, 2003).
The samples sprayed at 18 °C (E1) and 30 °C (E2) exhibited different
microbial growth (mesophilic and psychrotrophic bacteria), formation
of carbonyl groups, ORP, and thiols values, demonstrating that the
temperature of EWs can affect the process. In PC2, a high correlation of
TBARs and pH was observed up to 29 days of storage. The high tem-
perature associated with low pH values (5.55 to 5.92) during storage
favored lipid oxidation (Cheah & Ledward, 1996). When comparing
both experiments, it is evident the lower efficiency of spraying at a
higher temperatures (Fig. 4B), when compared to E1 (Fig. 4A).
Similar results were observed in the treatments using the combi-
nations BEW + AEW and BEW + SAEW and control at 30 psi (E3,
Fig. 4C) and 45 psi (E4, Fig. 4D), as a function of the storage time.
Microbial growth, increased color parameters and carbonyl groups, and
reduction of the variables ORP, TBARS, and thiols were the main factors
responsible for the higher variance in PC1. In PC2, it was possible to
visualize the discrimination between treatments and control in the ex-
periment E4 (45 psi, Fig. 4D), with higher pH values for the control, and
lower brightness for the treatments BEW + AEW and BEW + SAEW
during storage. On the other hand, in the experiment E3 (30 psi), no
difference was observed between treatments and control for these
variables.
D.R. Athayde et al.
4. Conclusions
The chlorine concentration of AEW and SAEW same after spraying
could be useful to exert a hygienizing action. The different compounds
in EW and pH and ORP values did not affect the pH, ORP, and red color
of pork loin. The combination BEW + AEW exhibited the highest an-
timicrobial effect among the conditions tested, and did not influence
the formation of carbonyl compounds and the oxidation of thiol groups,
regardless of the pressure and temperature used. Despite the anti-
oxidant effect of BEW on protein oxidation, the behavior of electrolyzed
water on the TBARS values was not clear. Our findings demonstrated
the possibility of using a smaller volume of EW for cleaning pork loins
after toileting, thus further studies are needed on the combination of
different types of EW, time, temperature, and pressure applied.
Acknowledgment
The authors are grateful to Coordenação de Aperfeiçoamento de
Pessoal de Nível Superior (CAPES) for scholarship to D. R. Athayde.
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