Postharvest Biology and Technology 156 (2019) 110953

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Postharvest Biology and Technology

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Effects of under-water cutting treatments on oleocellosis development, T

quality and shelf-life of minimally processed Persian lime fruit
Soriano-Melgar Lluvia de Abril Alexandraa, Raddatz-Mota Deniseb,
Díaz de León-Sánchez Fernandob, Rodríguez-Verástegui Lizette Lilianac,
⁎
Rivera-Cabrera Fernandob,
a
Cátedras-CONACYT, Centro de Investigación en Química Aplicada, Departamento de Plásticos en la Agricultura, Blvd. Enrique Reyna Hermosillo No.140, C.P. 25294,
Saltillo, Coahuila de Zaragoza, Mexico
b
Laboratorio de Postcosecha de Recursos Fitogenéticos y Productos Naturales, Departamento de Ciencias de la Salud, Universidad Autónoma Metropolitana-Iztapalapa,
CDMX, 09340, Mexico
c
Departamento de Biotecnologia, Universidad Autónoma Metropolitana-Iztapalapa, CDMX, 09340, Mexico

A R T I C LE I N FO A B S T R A C T

Keywords: The main problem for minimally processed acid limes commercialization is oleocellosis, a physiopathy generated
Calcium lactate on the surface of the fruit by the release of essential oils from flavedo’s oil glands due to processing, which causes
4-Hexylresorcinol sinking of the tissue, necrosis and loss of product quality. In order to reduce this problem, we evaluate the effect
Oil spotting of underwater cutting (UWC), either alone or in combination with other chemicals treatments on the devel-
Citrus latifolia
opment of oleocellosis and quality of minimally processed Persian lime.
Fresh-cut
Lipid peroxidation
Persian lime fruit was cut into 8 wedges inside a refrigerated chamber at 10 °C under water (UWC) at 4 °C
(control) or UWC with calcium lactate (5%) and 4-hexylresorcinol (4HR, 1 mM), either alone or in combination.
The obtained minimally processed Persian lime fruit were washed, disinfected and stored in crystal clear
polystyrene clam shells at 7.5 °C for 10 d. Visual quality and physicochemical parameters were determined.
UWC + calcium presented the lowest levels of dehydration, weight loss, lipid peroxidation and the minimum
damage at the cutting surface, showing to be the best treatment for minimally processed Persian lime reaching
10 d of shelf life and retaining marketable quality.

1. Introduction
Minimally processed fruit and vegetables are prepared by a series of
operations such as selection, washing, peeling, cutting and / or slicing,
disinfestation, draining and packaging (Tapia et al., 2015). The purpose
of minimally processed products (MPP) is to provide the consumer with
a vegetable product very similar to the fresh one, with a prolonged shelf
life, guaranteeing food safety and maintaining nutritional and sensory
quality. However, processing induces stress due to wounding and cell
damage, increasing respiratory rate, oxidative stress, loss of water and
firmness and production of anaerobic metabolites, which lead to loss of
quality (Toivonen and Brummell, 2008). Post-harvest technologies used
to extend MPP shelf life combine application of natural additives and
refrigeration (Bansal et al., 2015). Some of the used additives are
browning inhibitors such as cysteine, 4-hexylresorcinol (4HR), ascorbic
acid and citric acid, among others and calcium solutions that reduce the
damage on the cutting surfaces (Ghidelli et al., 2014; Pace et al., 2015;
Jideani et al., 2017). These treatments combined with refrigeration are
the most common post-harvest technology used to minimize the effects
of cutting on MPP (Saavedra del Águila et al., 2006). Storage tem-
perature range recommended for maintaining MPP quality, safety and
prolonging shelf life is between 0 and 5 °C (Cantwell and Suslow, 2002;
Oliveira et al., 2015). However, tropical or subtropical fruit such as
Persian lime, when stored below critical temperatures are (5 °C) de-
velop cold damage, which decreases fruit quality (Osorio-Mora and
Zacarias, 2000).
The main problem for Persian lime commercialization as an MPP is
oleocellosis, a physiopathy generated on the surface of the fruit by the
release of the essential oils from the oil glands of the flavedo (peel) due
to the processing, causing tissue sinking, necrosis and loss of product
quality. Several factors can increase the appearance of oleocellosis; in
order of importance, it can be mentioned: 1) turgor and initial fruit

⁎
Corresponding author.
E-mail addresses: alexandra.soriano@ciqa.edu.mx (L.d.A.A. Soriano-Melgar), deniseraddatz@gmail.com (D. Raddatz-Mota),
fdls5@yahoo.com.mx (F. Díaz de León-Sánchez), lizette.liliana@gmail.com (L.L. Rodríguez-Verástegui), frc@xanum.uam.mx (F. Rivera-Cabrera).

https://doi.org/10.1016/j.postharvbio.2019.110953
Received 16 April 2019; Received in revised form 9 July 2019; Accepted 9 July 2019
Available online 19 July 2019
0925-5214/ © 2019 Elsevier B.V. All rights reserved.
L.d.A.A. Soriano-Melgar, et al. Postharvest Biology and Technology 156 (2019) 110953

quality, 2) storage temperature and 3) type and conditions of the cut

(Bakkali et al., 2008). The essential oils that cause oleocellosis in citrus
fruit have been already identified; some of them are alcohols (geraniol,
linalool and nerol) and aldehydes (citral) (Eaks, 1968; Eckert and Eaks,
1989; Knight et al., 2002).
Alcohols and aldehydes are soluble in water; therefore, it has been
suggested that cutting under water could reduce the appearance of
oleocellosis compared to using a traditional cutting procedure in
Persian lime as MPP. Rivera-Cabrera et al. (2006) evaluated the effect
of the type of cut (traditional or underwater cutting) and storage tem-
perature (5 and 7.5 °C) on oleocellosis development and overall quality
of fresh-cut Persian lime, finding a lower development of oleocellosis
and a higher quality in the wedges obtained with underwater cutting
stored at 7.5 °C.
As mentioned above, there are several reports about the application
of postharvest treatments to reduce the cutting damage and browning
in MPP. However, this is the first work that evaluates the use of ad-
ditives to minimize the effects of cutting and avoid oleocellosis devel-
opment in minimally processed Persian lime. In Mexico, 90% of Persian
lime (Citrus latifolia T.) is exported and the rest is used for internal fresh
consumption. Persian lime as an MPP might represent a new marketing
alternative. The objective of this work was to evaluate the effect of
underwater cutting with calcium lactate and 4-hexylresorcinol, used
either alone or in combination on the development of oleocellosis and
minimally processed Persian lime quality.

2. Materials and methods

2.1. Plant material and experimental set-up

Persian lime fruit (Citrus latifolia T.) were obtained from experi-
mental orchard INIFAP-Ixtacuaco, Tlapacoyan, Veracruz in 2014; from
the main producing state in Mexico. Fruit was transported at room
temperature to the facilities of Universidad Autonoma Metropolitana in
Mexico City the same day of harvest. During the second day, fruit were
selected by color and size (dark green and a diameter of 4.2–5.5 cm),
obtaining four batches of 45 uniform and flawless fruit plus the initial
analysis. Each of the batches was randomly assigned to the different
treatments. Each fruit was cut into 8 wedges, eliminating both ends,
inside a refrigerated chamber at 10 °C. In a first experimental stage, the
studied treatments were: traditional cutting (TC) and underwater cut-
ting (UWC) at 4 °C. In a second stage the treatments evaluated were: 1)
underwater cutting at 4 °C (UWC), 2) UWC added with 5% (w/v) cal-
cium lactate (UWC + calcium), 3) UWC added with 1 mM 4-hex-
ylresorcinol (4HR) (UWC + 4HR), 4) UWC added with 5% (w/v) cal-
cium lactate and 1 mM 4HR (UWC + calcium + 4HR). All wedges
obtained from each treatment were washed with cold distilled water
(4–5 °C) containing 0.1 g L−1 of sodium hypochlorite (NaClO) for
2 min, and then rinsed for 1 min with cold distilled water (4–5 °C),
drained and the excess water was removed with absorbent paper. The
resulting samples were packed in crystal clear polystyrene clam shells
and stored at 7.5 °C ± 1 °C. Samples of 8 lime wedges (experimental
unit) in triplicate were taken from each treatment at each sampling day
(0, 2, 4, 6, 8 and 10 d).
2.2. Determination of visual quality parameters
Visual quality parameters were determined for each sample before
using them for the physical and chemical quality attributes analysis.
For evaluation of visual quality of minimally processed Persian lime
fruit, incidence and severity of dehydration and the development of
oleocellosis were evaluated using hedonic scales described in Fig. 1A
and B with the following categories: 1) no damage, 2) light damage, 3)
moderate damage (acceptable limit of consumption), 4) severe damage
and 5) extreme damage. All evaluations were made by the same person
(the first author).
Fig. 1. Scales used to evaluate the A) dehydration and B) oleocellosis devel-
opment on segments of minimally processed Persian lime fruit stored at 7.5 °C
and C) histological section (40×) stained with safranin-fast green in the cut
region of minimally processed Persian lime fruit (initial analysis).
Hedonic scale: 1) no damage, 2) light damage, 3) moderate damage (acceptable
limit of consumption), 4) severe damage and 5) extreme damage. Abbreviations:
E = epidermis; O = oil glands; A = albedo or mesocarp; F = flavedo or exo-
carp. (For interpretation of the references to colour in this figure legend, the
reader is referred to the web version of this article.)
2.3. Physical and chemical quality attributes analysis
Pulp color was determined with a manual colorimeter CR-400/410,
Konica Minolta, New Jersey, USA. Subsequently, the juice from 8
wedges was used to determine titratable acidity (TA), pH and total
soluble solids (TSS). The juice obtained was weighed and used to cal-
culate juice percentage. TA was determined using phenolphthalein
(0.1%, w/v) as indicator and results were reported as percentage of
citric acid. pH was quantified with a potentiometer (Conductronic pH
10, Labequim SA de CV, Mexico). TSS were measured with a manual
refractometer (Atago Co. Ltd., Tokyo, Japan) and reported as percen-
tage.
2.4. Weight loss (WL)
To determine percentage of weight loss (WL), samples of 8 wedges
in triplicate were used for each treatment. Initial weight and cumulative
weight loss at each sampling date was recorded for each fruit sample,
using an analytical balance (OHAUS, GT4100, Florham Park, New
Jersey, USA). Results were expressed as percentage of weight loss and
were calculated using the following formula:
Weight loss percentage (%) = (1 – Wn / Wi) (100)
Where:
Wn=Fruit weight (g) at day n.

2
L.d.A.A. Soriano-Melgar, et al. Postharvest Biology and Technology 156 (2019) 110953

Wi=Fruit initial weight (g).

2.5. Determination of total phenolic compounds and antioxidant capacity

Total phenolic compounds content was determined using the Folin-

Ciocalteu reagent according to Singlenton and Rossi (1965). One mil-
liliter of lime juice was diluted with 80% aqueous methanol and used
for the reaction mixture. Absorbance was measured at 765 nm. Total
phenolic compounds content was expressed as gallic acid equivalent in
g L−1 of lime juice (g L−1 GAE).
Antioxidant capacity was determined according to the method de-
scribed by Brand-Williams et al. (1995). Decrease in absorbance at
515 nm was measured and results were expressed as antioxidant ca-
pacity in terms of ascorbic acid equivalent (AAE) in mol L−1 of lime
juice (mol L−1 AAE).

2.6. Lipid peroxidation

Lipid peroxidation was determined in the flavedo using the method

described by Rivera et al. (2007). Lipid peroxidation assay is based on
quantification of malondialdehyde (MDA), the main product of mem-
brane polyunsaturated fatty acid peroxidation (Buege and Aust, 1978).
Lipid peroxidation detected at 586 nm was reported as millimoles of
MDA per kilogram of fresh weight (mmol kg−1 MDA).

2.7. Determination of anaerobic metabolites (acetaldehyde and ethanol)

Five milliliters of juice were mixed with 2 g of NaCl in a vial, which

was sealed and incubated for 10 min at 35 °C, and then, stirred for 30 s.
A sample of 1 mL of the gas phase of the headspace was collected and
injected into a gas chromatograph (Hewlet Packard 5890 serie II, Palo
Alto, USA). Analyses were carried out on a packed Column (Carbowax
at 5% on carbopack 60/80) using helium as the carrier gas. The flow Fig. 2. First experimental stage: A) Dehydration, B) development of oleocellosis
rate through the column was 20 mL min−1 and the column temperature in minimally processed Persian lime stored at 7.5 °C.
was set at 80 °C, then raised to 130 °C at 10 °C min−1 and the final The dashed line indicates the acceptable limit for consumption. Each value is
temperature was maintained at 130 °C for 1 min. Injector and detector the average of three replicates (8 wedges) ± standard deviation. Hedonic scale:
temperatures were 115 °C and 200 °C, respectively (Pelayo, 2001). 1) no damage, 2) light damage, 3) moderate damage (acceptable limit of con-
sumption), 4) severe damage and 5) extreme damage. Abbreviations: TC (tra-
ditional cutting), UWC (under-water cutting).
2.8. Evaluation of the effects of cutting by light microscopy

Samples of 8 wedges in triplicate for each treatment were fixed with
FAA (formaldehyde, ethanol 96° v/v, glacial acetic acid and distilled
water) during 48 h (Gray, 1964) and dehydrated using 8 series of
ethanol and tert-butyl alcohol (TBA) at different percentages for 5 h for
each series. Then, the tissue was embedded in paraffin 48 h before
sectioning (Gray, 1964). Histological sections of ten microns were made
in a rotating microtome and dyed with safranin/fast green and naphthol
blue black. Images were obtained in microphotography (López-Curto
et al., 2005).
2.9. Statistical analysis
Analysis of variance (ANOVA) followed by Tukey’s multiple means
comparison test was performed. The level of statistical significance was
α = 0.05. The results of determination of visual quality parameters
(incidence and severity) were analyzed using Kruskal-Wallis multiple
means comparison test and Dunn's multiple comparison test. For all
cases, a significance level of α = 0.05 was used. All data were analyzed
using the NCSS software (Hintze, 2007).
3. Results and discussion
3.1. Effects of under-water cutting treatments on visual quality parameters
In MPP, by cutting, the natural protection of the epidermis is re-
moved, generating a greater tissues surface exposure and therefore,
greater dehydration and weight loss. (Artés-Hernández et al., 2007;
Colleli and Amodio, 2016). As mentioned in Section 2.1 during a first
experimental stage, the effect of TC and UWC procedures on the overall
quality of the segments of minimally processed Persian lime were
evaluated. Fig. 2 shows the results obtained for incidence and severity
of dehydration and development of oleocellosis. TC increased symp-
toms of dehydration and development of oleocellosis after 2 days of
storage and showed the highest values during the entire storage period
compared to the UWC procedure. In both parameters, dehydration and
oleocellosis, values observed with the TC procedure reached extreme
damage towards the end of storage.
Results obtained during this experimental phase showed that UWC
reduce dehydration symptoms and development of oleocellosis in
minimally processed Persian lime stored at 7.5 °C.
During a second experimental stage, the effect of underwater cut-
ting with calcium lactate and 4-hexylresorcinol, used either alone or in
combination on the development of oleocellosis, quality and shelf-life
of minimally processed Persian lime fruit, was evaluated. In Fig. 3,
values of dehydration for all treatments stored at 7.5 °C are shown,
which remained within the acceptable limit for consumption during the
first six days of storage according to the used hedonic scale (Fig. 1A).
Fruit treated with UWC and UWC + 4HR had higher values of dehy-
dration exceeding the limit established as acceptable (Fig. 1A) from the
eighth day onward (Fig. 3A). The best results after 10 d of storage were
obtained for the UWC + calcium and UWC + calcium + 4HR treat-
ments (Fig. 3A).

3
L.d.A.A. Soriano-Melgar, et al.
Fig. 3. A) Dehydration, B) oleocellosis development and C) weight loss per-
centage in minimally processed Persian lime stored at 7.5 °C.
The dashed line indicates the acceptable limit for consumption. Each value is
the average of three replicates (8 wedges) ± standard deviation. Hedonic scale:
1) no damage, 2) light damage, 3) moderate damage (acceptable limit of con-
sumption), 4) severe damage and 5) extreme damage. Abbreviations: UWC
(under-water cutting), UWC + calcium [UWC added with 5% (w/v) calcium
lactate], UWC + 4HR (UWC added with 1 mM 4-hexylresorcinol),
UWC + calcium+4HR [UWC added with 5% (w/v) calcium lactate and 1 mM
4-hexylresorcinol]. Different letters indicate the statistically significant differ-
ence among treatments for the same storage day (P < 0.05) and ns indicate no
significant differences.
Oleocellosis development showed the same tendency as the afore-
mentioned results for dehydration (Fig. 1B). That means that oleo-
cellosis was within the acceptable limit for consumption for all the
treatments (Fig. 1B) until the sixth day of storage (Fig. 3B). The treat-
ment that showed lower oleocellosis in fruit stored at 7.5 °C was
UWC + calcium + 4HR (Fig. 3B). Treatments with calcium lactate and
4
Postharvest Biology and Technology 156 (2019) 110953
4HR have been widely used as pre and post-harvest treatments, being
effective in maintaining quality (Barzegar et al., 2018). The best results
observed in dehydration levels and in the development of oleocellosis in
fruit treated with UWC + calcium and UWC + calcium + 4HR could be
due to the direct effect of calcium lactate and not to 4HR treatment
(Fig. 3A and B). This might be due to the action of calcium ions in-
teracting with the cell wall and middle lamella pectin, stabilizing cell
membrane and favoring cell turgor pressure (Luna-Guzmán and Barrett,
2000).
On the other hand, 4HR is a small organic compound that is used as
antiseptic and antioxidant, but its molecular properties have not been
clearly elucidated (Kim et al., 2019). In this sense, a possible explana-
tion for the lower oleocellosis development in fruit treated with
UWC + calcium + 4HR could be due a synergistic effect of both ad-
ditives (Fig. 3B).
3.2. Effects of under-water cutting treatments on physical and chemical
quality attributes
3.2.1. Color parameters
Changes in appearance and texture are the most important char-
acteristics for acceptance of MPP; within the appearance, color is a very
important parameter (Rivera-López et al., 2005). No significant differ-
ences (P > 0.05) were observed for the different color parameters,
luminosity (L*), chromaticity (C*) and Hue angle (h) in the pulp of
minimally processed Persian lime fruit among treatments and days of
storage (Fig. 4). In the literature, it has been reported that the decrease
in luminosity (L*) and the variation in Hue (h) in MPP have been as-
sociated with oxidative browning and water loss (Portela and Cantwell,
2001; Rivera-López et al., 2005; Rojas-Graü et al., 2006; Teixeira et al.,
2007). Our results showed little variation in the color parameters (L*
and h) (Fig. 4), which could indicate the absence of browning and the
preservation of the visual quality and appearance of the fruit (Figs. 3A,
B and 4).
3.2.2. TA, pH, TSS and juice content
Titratable acidity (TA) is a quantitative measure of the organic acids
present in the fruit; in general, it decreases during maturation, senes-
cence and under storage conditions (Gorny and Kader, 2005). TA re-
sults did not show significant differences (P > 0.05) among treatments
and storage days (Table 1). TA values in MPP Persian lime were in a
range of 5–7 % citric acid, which is in accordance with percentages
reported for Persian lime (Hanif et al., 2017; Machado et al., 2017).
pH as a physicochemical parameter is important because it is related
to the content of organic acids in fruit and vegetables (Soliva-Fortuny
and Martín-Belloso, 2003). pH values did no show significant differ-
ences (P > 0.05) among treatments (Table 1) and oscillated in a range
of 2.1–2.3, which are similar to the values reported by Artés-Hernández
et al. (2007) in fresh-cut 'Lisbon' lemons. These results are important,
considering that in MPP, an acid pH decreases the development of
microorganisms (Soliva-Fortuny and Martín-Belloso, 2003). On the
other hand, it is considered that high pH values (> 4.6) in MPP result in
more perishable products (Wiley, 1997).
Sugars are the soluble solids that are found in a greater proportion
in the juice of fruit and therefore, TSS content is often used as criteria of
maturity for harvesting (Kitinoja and Kader, 2003). In general, TSS
values showed no significant differences (P > 0.05) among treatments
and storage days (Table 1). TSS values (7.2–9 %) were maintained si-
milar to the initial analysis after 10 d of storage in all treatments
(Table 1). Artés-Hernández et al. (2007) reported values of 7–7.6 % of
TSS in minimally processed lemons that did not change during the
storage time and temperatures used.
Two of the most important quality attributes in Persian lime are
juice content and size; a minimum of 45% juice per weight and a size of
50–63 mm is required according to the Mexican Quality Standard for
Persian Lime NMX-FF-077-1996 (Rivera-Cabrera et al., 2010). In
 (Table 1).
lime fruit stored at 7.5°C.
L.d.A.A. Soriano-Melgar, et al.

3.3. Percentage of weight loss

and ns indicate no significant differences.
lactate and 1 mM 4-hexylresorcinol]. Different letters indicate the statistically
Each value is the average of three replicates (8 wedges) ± standard deviation.

Artés et al., 2007; Toivonen and Brummell, 2008). Weight loss (WL)
Minimally processed products are highly susceptible to dehydration
5% (w/v) calcium lactate], UWC + 4HR (UWC added with 1 mM 4-hex-

general, in this study values of juice percentage were maintained within

significant difference among treatments for the same storage day (P < 0.05)

rapid loss of water, reducing fruit weight and quality (Brecht, 1995;
membranes by the wound generated during processing which causes a
ylresorcinol), UWC + calcium+4HR [UWC added with 5% (w/v) calcium

and solute leaching or leakage, due to the damage caused to the cell
Fig. 4. Color parameters (L*, C*, h) in the pulp of minimally processed Persian

(P > 0.05) among treatments during the entire storage period

a range of 35–45 % of juice and did not show significant differences
Abbreviations: UWC (under-water cutting), UWC + calcium [UWC added with

5
Table 1
Physical and chemical quality attributes in Persian lime fruit minimally processed stored at 7.5°C.
Parameters Initial Days of storage

2d 4d 6d 8d 10 d

UWC UWC + UWC UWC + UWC UWC + UWC + UWC + UWC UWC + UWC + UWC + UWC UWC + UWC UWC + UWC UWC + UWC UWC +
calcium + calcium calcium 4HR calcium calcium 4HR calcium calcium + calcium calcium + calcium
4HR + 4HR + 4HR + 4HR 4HR + 4HR 4HR + 4HR

TA (% citric 6.68 6.31 6.57 ± 6.53 6.89 ± 5.82 5.55 ± 5.97 ± 5.87 ± 5.72 5.91 ± 6.31 ± 6.02 ± 6.78 6.02 ± 5.74 6.04 ± 6.10 6.49 ± 5.76 5.87 ±
acid) ± 0.4- ± 0.1- 0.20 aAB ± 0.1- 0.59 aA ± 0.1- 0.41 aB 0.21 aAB 0.30 aA ± 0.1- 0.10abAB 0.20 aAB 0.23 abA ± 0.5- 0.59 aAB ± 0.3- 0.21 aA ± 0.4- 0.51 aAB ± 0.1- 0.43 aA
4 aAB 3 aAB 3 aA 7 aAB 0 bB 6 aA 0 aB 5 aAB 7 aB
pH 2.09 2.28 2.21 ± 2.17 2.16 ± 2.32 2.23 ± 2.17 ± 2.24 ± 2.23 2.13 ± 2.15 ± 2.21 ± 2.31 2.28 ± 2.21 2.25 ± 2.30 2.23 ± 2.18 2.20 ±
± 0.0- ± 0.0- 0.05 ± 0.0- 0.07 bAB ± 0.0- 0.03 bAB 0.01 bA 0.03 abA ± 0.0- 0.04 bBC 0.03abAB 0.02 aA ± 0.0- 0.02 abA ± 0.0- 0.03 abA ± 0.0- 0.05abAB ± 0.0- 0.01 bA
abAB
4 aC 2 aAB 1 abA 5 aA 2 aBC 4 aAB 4 bA 4 aAB 3 bA
TSS (%) 7.73 7.93 8.27 ± 8.13 8.33 ± 7.60 8.47 ± 8.13 ± 8.87 ± 7.40 8.07 ± 8.47 ± 8.80 ± 7.60 8.00 ± 8.13 8.40 ± 8.07 8.07 ± 8.73 8.53 ±
± 0.1- ± 0.1- 0.23 aA ± 0.1- 0.23 aAB ± 0.2- 0.31 abA 0.12bcBC 0.31 aA ± 0.2- 0.12 bAB 0.31abAB 0.20 aA ± 0.4- 0.20 aAB ± 0.1- 0.72 aAB ± 0.2- 0.12 bAB ± 0.3- 0.12abAB
2 aAB 2 aAB 2 aBC 0 cAB 0 cB 0 aAB 2 aBC 3 bA 1 aA
Juice content 40.72 35.44 38.52 35.81 35.99 ± 36.79 38.12 37.23 ± 38.85 ± 36.25 35.48 ± 36.45 ± 40.19 ± 31.88 34.12 34.83 37.36 ± 31.25 34.53 ± 33.74 39.55 ±
(%) ± 0.6- ± 3.9- ± 3.24 ± 2.1- 2.07 aA ± 2.2- ± 1.86 8.06 aA 6.40 aA ± 3.1- 0.81 aB 1.92 aA 4.76 aA ± 2.2- ± 0.19 ± 1.4- 0.42 aA ± 4.8- 2.33 aB ± 1.4- 3.99 aA
aAB aAB abB
4 aA 0 aAB 3 aA 8 aAB 6 aAB 3 bB 1 abA 9 aB 6 aA

Same letters, lower case for treatments and upper case for days of storage for each parameter, do not differ significantly (p < 0.05) according to Tukey's test. Each value is the average of three replicates (8
wedges) ± standard deviation. Abbreviations: d (days), UWC (under-water cutting), UWC + calcium [UWC added with 5% (w/v) calcium lactate], UWC + 4HR (UWC added with 1 mM 4-hexylresorcinol), UWC + calcium
+4HR [UWC added with 5% (w/v) calcium lactate and 1 mM 4-hexylresorcinol].
Postharvest Biology and Technology 156 (2019) 110953
L.d.A.A. Soriano-Melgar, et al. Postharvest Biology and Technology 156 (2019) 110953

increased throughout the storage period in all treatments (Fig. 3C).

Significant differences (P < 0.05) were observed in WL after six days
of storage in treatments UWC and UWC + 4HR compared with treat-
ment UWC + calcium (Fig. 3C). Fruit treated using UWC + calcium
showed the lowest WL (2.83%) after 10 d of storage (Fig. 3C). Barry-
Ryan and O’Beirne, 1998 mention that WL is related to the degree of
physiological damage and the increase in the rate of senescence, de-
creasing MPP quality. WL between 4–8 % has negative effects on ap-
pearance and acceptance of MPP (Rivera-López et al., 2005). Our re-
sults are consistent with those reported in the literature, since the
maximum percentage of WL observed did not exceed 3.7% after 10 d of
storage (Fig. 3C).

3.4. Effects of under-water cutting treatments on total phenolic compounds,

antioxidant capacity and lipid peroxidation

Phenolic compounds, ubiquitous in plants, are an essential part of

the human diet, and are of considerable interest due to their antioxidant
properties. For this reason, maintenance of phenolic compounds in the
MPP plays an important role for their nutritional value. In general, no
significant differences (P > 0.05) were observed in total phenolic
compounds content among treatments (Fig. 5A). Total phenolic com-
pounds content did not change after 10 d of storage compared to the
initial content (Fig. 5A). These results are consistent with those re-
ported in the literature indicating that the maintenance of phenolic
compounds levels in MPP is an indicator of the conservation of fruit
nutritional quality (Soliva-Fortuny and Martín-Belloso, 2003).
Antioxidant capacity is a parameter generally determined as part of
the phytochemical profile of fruit tissues and represents a quick mea-
sure of their antioxidant content. In this study, no significant differ-
ences were observed (P > 0.05) in the antioxidant capacity among
treatments (Fig. 5B). In general, total antioxidant capacity was main-
tained constant during the 10 d of storage in all treatments, obtaining
values similar to the initial analysis (33.53 mol L−1) (Fig. 5B). Artés-
Hernández et al. (2007) reported similar values of antioxidant capacity
in minimally processed lemons. Our results and those reported by Artés-
Hernández et al. (2007), suggest that wound stress caused by cutting
did not significantly affect the nutritional quality of minimally pro-
cessed limes and lemons after 10 d of storage.
It has been reported that stress by wounding accelerates maturation
and senescence in MPP, increasing texture loss and softening of tissues
(Artés-Hernández et al., 2007; Toivonen and Brummell, 2008). An in-
direct way to determine the damage caused by cutting is by quantifying
lipid peroxidation levels as an indicator of membrane damage. Sig-
Fig. 5. A) Total phenolic compounds contents, (B) antioxidant capacity
nificant differences in lipid peroxidation levels were observed among
equivalents and (C) lipid peroxidation in the peel of minimally processed
treatments. Fruit treated with UWC + calcium showed the lowest levels
Persian lime stored at 7.5°C.
of lipid peroxidation, while levels in fruit treated with UWC (control)
Each value is the average of three replicates ± standard deviation.
were maintained similar to initial analysis along storage period Abbreviations: UWC (under-water cutting), UWC + calcium [UWC added with
(Fig. 5C). Fruit treated with UWC + calcium + 4HR and UWC + 4HR 5% (w/v) calcium lactate], UWC + 4HR (UWC added with 1 mM 4-hex-
showed the highest levels of lipid peroxidation compared to UWC ylresorcinol), UWC + calcium+4HR [UWC added with 5% (w/v) calcium
(control); the increase on lipid peroxidation levels on these treatments lactate and 1 mM 4-hexylresorcinol]. Different letters indicate the statistically
was maintained throughout storage (Fig. 5C). However, the lipid per- significant difference among treatments for the same storage day (P < 0.05)
oxidation levels were lower for fruit treated with UWC + calcium + and ns indicate no significant differences.
4HR than for fruit treated with UWC + 4HR treatment, which could
indicate that calcium lactate can decrease membrane damage (lipid biosynthesis (Xin-Shi et al., 2007). Our results showed that acet-
peroxidation) or that 4HR treatment increase it. Our results, as well as aldehyde levels in fruit treated with UWC and UWC + calcium main-
several studies, show a beneficial effect of the application of calcium for tained similar values to the initial one during the 10 d of storage
maintaining minimally processed fruit quality (Beirão-da-Costa et al., (Fig. 6A). On the other hand, in fruit treated with UWC + 4HR and
2008; Shaarawi et al., 2016; Artés-Hernández et al., 2017). UWC + calcium + 4HR, acetaldehyde levels decreased from the fourth
day of storage onward, except at 10 d in fruit treated with
3.5. Effect of under-water cutting treatments on anaerobic metabolites UWC + calcium + 4HR, where an increase was observed (P < 0.05) in
comparison with fruit treated with UWC + 4HR.
Perception of undesirable flavors and odors in citrus fruit is asso- This decrease might be explained by an increase on alcohol dehy-
ciated with acetaldehyde and ethanol accumulation. In general, acet- drogenase (ADH) activity which would result in an increase in ethanol
aldehyde and ethanol normally accumulate at low levels during fruit levels. However, there were no significant differences in ethanol levels
ripening and play an important role in aromatic compounds

6
L.d.A.A. Soriano-Melgar, et al.
Fig. 6. Anaerobic metabolites: A) acetaldehyde and B) ethanol in juice of
minimally processed Persian lime fruit stored at 7.5°C.
Each value is the average of three replicates ± standard deviation.
Abbreviations: UWC (under-water cutting), UWC + calcium [UWC added with
5% (w/v) calcium lactate], UWC + 4HR (UWC added with 1 mM 4-hex-
ylresorcinol), UWC + calcium+4HR [UWC added with 5% (w/v) calcium
lactate and 1 mM 4-hexylresorcinol]. Different letters indicate the statistically
significant difference among treatments for the same storage day (P < 0.05)
and ns indicate no significant differences.
among treatments with 4HR (Fig. 6B). Another possible explanation
could be low activity of the pyruvate decarboxylase (PDC) enzyme
which catalyzes acetaldehyde formation from pyruvate. However, in
this work none of these two enzymatic activities were determined.
Acetaldehyde levels in citrus fruit range between 0.15–12 μg L−1
(Shaw, 1991). In the present work, acetaldehyde levels range from 2
to15 μg L−1, which are within the limits of perception (Fors, 1988). In
general, significant differences (P < 0.05) were observed from the
sixth day of storage onward in ethanol levels among treatments.
An increment in ethanol levels was observed in fruit treated with
UWC + calcium after 10 d of storage compared to the other treatments
used (Fig. 6B). However, the observed values are within the range re-
ported in the literature. Ethanol levels in this work were within the
range of 24–160 μg L−1 and values reported in citrus juice as aroma
volatile are in the range of 53–380 μg L-1 (Shaw, 1991).
3.6. Evaluation of under-water cutting on Persian lime fruit by light
microscopy
Histochemical analysis was performed to evaluate the damage
generated at the cutting surface in minimally processed Persian lime, as
well as the effect of the treatments used in the present work.
Dyes used were safranin-fast green (Fig. 7A) and naphthol blue
black (Fig. 7B) which allowed to compare the primary and secondary
cell walls, as well as membrane components, respectively.
7
Postharvest Biology and Technology 156 (2019) 110953
In Fig. 7, images from the cutting section for the different treat-
ments at 2, 6 and 10 d of storage are compared using the initial analysis
as reference.
The greatest damage in the cell walls of the cutting region was
observed in fruit treated with UWC + 4HR and UWC + calcium + 4HR
from the sixth day of storage onward (Fig. 7A). The same effect was
observed with naphthol blue black dye for these treatments from the
second day onward (Fig. 7B); however, damage was observed for all
treatments.
These results coincide with the increase in dehydration and oleo-
cellosis observed at 8 and 10 d of storage in fruit treated with
UWC + 4HR (Fig. 3A and B). In addition, the highest levels of lipid
peroxidation (membrane damage) were observed in fruit treated with
UWC + 4HR and UWC + calcium + 4HR during the entire storage
period (Fig. 5C). On the other hand, the best results were obtained in
fruit treated with UWC and UWC + calcium showing the least damage
at the cutting region in cell walls (safranin-fast green) and in the
components of cell membranes (naphtol blue black) throughout storage
(Fig. 7).
The beneficial effect of UWC + calcium treatment observed using
histochemical dyes is consistent with the decrease in dehydration and
lipid peroxidation levels (Figs. 3A and 5 C). This decrease in dehydra-
tion resulted in a lower weight loss by the end of the storage period in
the fruit treated with UWC + calcium (Fig. 3C). These results indicate
that treatments with calcium help to improve appearance and visual
quality of minimally processed Persian lime fruit. Similar beneficial
effects have been reported for minimally processed melon, kiwi, pears
and strawberries, showing that application of different calcium treat-
ments improve appearance and texture (Luna-Guzmán and Barrett,
2000; Lamikanra and Watson, 2004; Beirão-da-Costa et al., 2008). This
beneficial effect of calcium has been associated to an increase in firm-
ness, by forming pectates of calcium that increase membrane rigidity
and slow down polygalacturonase enzyme activity, decreasing perme-
ability and stabilizing cell membranes, and therefore, avoiding tissues
water loss and increasing cell-cell adhesion (Tsantili et al., 2002;
Lamikanra and Watson, 2004; Aguayo et al., 2008).
4. Conclusions
Even though limes and lemons as minimally processed product
(MPP) are in high demand in hotels, restaurants and bars; in general,
they go unnoticed. Acid limes (Persian and Mexican lime) are preferred
by consumers since they are more aromatic compared to lemons. In this
work, we evaluate the use of post-harvest treatments for preserving
quality of Persian lime as an MPP.
Results from this study showed that oleocellosis continue being the
main problem for commercialization of Persian lime fruit as an MPP.
The use of UWC decreased levels of oleocellosis in minimally processed
Persian lime, preserving fruit quality. The lowest levels of oleocellosis
were observed for UWC + calcium treatment during the first 6 d and
UWC + calcium + 4HR during the whole period of storage. In addition,
no differences were observed in the evaluated physicochemical para-
meters (color, pH, TA and percentage of juice) among treatments,
meaning that there was no negative effect of the treatments on the
nutritional quality of minimally processed Persian lime fruit. Also, an-
tioxidant capacity and total phenolic compounds content after 10 d of
storage were similar to the values obtained in the initial analysis for all
treatments.
Results for weight loss, dehydration, lipid peroxidation and histo-
chemical analysis by light microscopy showed that the best treatment
for processing Persian lime fruit as a minimally processed product was
UWC + calcium reaching 10 d of shelf life retaining marketable quality.
Acknowledgements
This work was supported by Universidad Autónoma Metropolitana-
L.d.A.A. Soriano-Melgar, et al. Postharvest Biology and Technology 156 (2019) 110953

Fig. 7. Cross sections of the peel of minimally processed Persian lime fruit (10X) dyed with (A) safranin/fast green and (B) naphthol blue black. (For interpretation of
the references to colour in this figure legend, the reader is referred to the web version of this article.)
Abbreviations: d (days), UWC (under-water cutting), UWC + calcium [UWC added with 5% (w/v) calcium lactate], UWC + 4HR (UWC added with 1 mM 4-hex-
ylresorcinol), UWC + calcium+4HR [UWC added with 5% (w/v) calcium lactate and 1 mM 4-hexylresorcinol].
E=epidermis; O = oil glands; A=albedo or mesocarp; F=flavedo or exocarp.

Iztapalapa. We also thank Sergio Alberto Curti-Díaz of INIFAP for as- 18–22.
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