Food Research International 42 (2009) 511515

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Food Research International

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Edible lm based on candelilla wax to improve the shelf life and quality of avocado
Saul Saucedo-Pompa a, Romeo Rojas-Molina b, Antonio F. Aguilera-Carb b, Aide Saenz-Galindo a,
Heliodoro de La Garza a, Diana Jasso-Cant b, Cristbal N. Aguilar a,*
a
Department of Food Science and Technology, School of Chemistry, Universidad Autnoma de Coahuila, Blvd. Venustiano Carranza, 25280 Saltillo, Coahuila, Mexico
b
Universidad Autnoma Agraria Antonio Narro Calzada Antonio Narro 1923, Buenavista, Saltillo, Coahuila, Mexico

a r t i c l e i n f o a b s t r a c t

Article history: In this study the effect of addition of ellagic acid (at three different concentrations) into candelilla wax
Received 22 November 2008 matrix on shelf life and quality of whole avocados was studied. Control treatments were avocados coated
Accepted 18 February 2009 with candelilla wax without ellagic acid and avocados without coating. The fruits were chosen for their
maturity, size, free from infection and physical defects. All those samples were inoculated with a concen-
trated suspension of spores of Colletotrichum gloeosporioides, the main phytopathogenic fungus for avo-
Keywords: cados. Experiments were carried out completely divided into randomized groups. Changes in
Candelilla wax
appearance, solids content, pH, aw, lightness (L* value) and weight loss were monitored during 6 weeks
Colletotrichum gloeosporioides
Avocado
every 8 days. A sensory evaluation of avocados coated with the best edible lm was also performed. Edi-
Ellagic acid bel lms were able to reduce signicantly the damage caused by C. gloeosporioides, reducing also signif-
icantly the change in appearance and weight loss in the fruits. Use of ellagic acid as part of the edible lm
has an important effect to improve the quality and shelf life of avocado. With this work we found that
using this new protection system the negative effects of C. gloeosporioides can be successfully reduced.
2009 Elsevier Ltd. All rights reserved.

1. Introduction preservatives on the food surfaces (Oussallah, Caillet, Salmieri,

Saucier, & Lacroix, 2004). Several compounds have been proposed
In recent years, food packaging research has focused more on for antimicrobial activity in food packaging, including organic
biodegradable lms, including lms made from plant sources (Sey- acids, enzymes such as lysozyme, and fungicides such as beno-
dim & Sarikus, 2006). Because consumers demand less use of myl, imazalil and natural antimicrobial compounds such as spices
chemicals on minimally processed fruits and vegetables, more (Tharanathan, 2003a, b).
attention has been paid to search for naturally occurring sub- To develop these lms, it is necessary to consider the mechan-
stances able to act as alternative antimicrobials and antioxidants. ical, physical and chemical factors involved in the storage of fruits.
The ability of edible lms to retard moisture, oxygen, aromas and In addition, these lms can confer nutritional and organoleptic
solute transport may be improved by including additives such as properties to the food if added as antioxidants, colorants or arti-
antioxidants, antimicrobials, colorants, avors, fortifying nutrients cial avors (Saucedo-Pompa, Jasso-Cantu, Ventura-Sobrevilla,
and spices in lm formulation (Pranoto, Rakshit, & Salokhe, 2005). Senz-Galindo, & Aguilar, 2007).
Besides incorporation of antimicrobial agents into packaging mate- The use of edible lms cannot replace the use of traditional
rials has also become very popular since oxidation is a major prob- packaging materials, but it is necessary to take into account their
lem affecting the food quality (Mecitou-Gbilmez, Yemeniciou, functional characteristics and the possible advantages in certain
& Arslanou, 2007). applications (Baldwin, Medlar-Carriedo, & Baker, 1995). The inu-
Antimicrobial packaging is a form of active packaging that ence of an additive will depend on their concentration, chemical
could extend the shelf life of food products while providing structure, degree of dispersion in the lm and degree of interaction
microbial safety for consumers (Rooney, 1995). It acts by reduc- with the polymer (Kester & Fennema, 1986). If there is an adequate
ing, inhibiting, or retarding the growth of pathogen microorgan- homogenization of the system it is possible to ensure uniformity in
isms in packed foods and packaging material (Vermeiren, the size and distribution of particles dispersed phase, improving
Devlieghere, van Beest, de Kruiif, & Debevere, 1999). The coating the barrier functionality against the mass transfer of the formed
can serve as a carrier for antimicrobial compounds and/or antiox- lm (Bosquez-Molina, 2005).
idants compounds in order to maintain high concentrations of The candelilla wax is a complex substance of vegetable origin,
insoluble in water, but highly soluble in acetone, chloroform, ben-
zene and other organic solvents, is characterized by a high content
* Corresponding author. Tel.: +52 844 416 1238; fax: +52 844 415 9534.
E-mail address: [email protected] (C.N. Aguilar).
of hydrocarbons (around 50%) and a relatively low quantity of vol-

0963-9969/$ - see front matter 2009 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodres.2009.02.017
512 S. Saucedo-Pompa et al. / Food Research International 42 (2009) 511515

atile esters and is recognized by the FDA as a substance GRASS for balance [Ohaus mode E-02130, Pine Brook, NJ]. Humidity of 45%.
application in the foods industry (Saucedo Pompa, 2007). Measurement of the following characteristics was performed dur-
Tannins are secondary metabolites present in these plants. ing the storage period. They were conserved at 5 C by a period of 6
Some of the principal components of these substances are the ella- weeks and the physicochemical were evaluated every 8 days.
gic and gallic acids. Several medicinal and industrial applications The lightness of samples was analyzed using the L value in a
have been found for these acids such as antitumoral, antioxidant, portable colorimeter (ColorTec-PCM, Clifton, NJ). Changes in their
antimicrobial, antiviral and antiinammatory. In addition, some aspect were photographically recorded (with a HewlettPackard
of them may reduce heart disease (Khanbabaee & Van Ree 2001; Photosmart R927 digital camera with a 24 zoom, Palo Alto, CA).
Ventura et al., 2008). Weight loss was measured with an analytical balance (Ohaus mod-
The ellagic acid present in grapes, strawberries, blackberries, el E-02130). The solid contents were measured with a thermobal-
blueberries, nuts and other foods is a type of phenolic compound ance (Ohaus model MB-200). Firmness was measured with a
that acts as an antioxidant; however, they are not easily available Humboldt universal penetrometer (model H-1200, Chicago, IL).
and can vary in effectiveness depending on if they are used in their The pH values were obtained using a pH meter (Orion model-
natural form or puried. Such compounds may also protect against 420, Boston, MA). A slurry was made with a mix of fruit and distil-
ultraviolet light, attack by insects and microorganisms. In order to late water (1:10 w/v), stirring for 10 min, then an electrode was
be bioavailable, ellargic acid needs to be in a form under which the introduced to measure the pH value. Water activity (aw) was mea-
cell can recognize it and use it. This may be the free chemical form sured with an AquaLab Decagon (series 3, Pullman, WA).
or in a combined form with another biomolecule; in general, ellar-
gic acid combines itself with molecules of sugar (Vasconcellos, 2.3. Microbiological assays
1994).
Mexico is the largest producer of avocados in the world (Sauce- All treatments were inoculated with a suspension of 1  107
do-Pompa, Jasso-Cantu, Rojas-Molina, Senz-Galindo, & Aguilar, spores of C. gloeosporioides. They were conserved at 5 C by a period
2008), however this country faces several requests for approving of 6 weeks and were evaluated every 8 days to evaluate the effect
the exportation of the avocados to the United States and Europe. of antifungal edible coatings with the addition of antioxidant
In addition, the shelf life of these fruits is very short and their qual- additives.
ity is seriously affected by the attack of phytopathogenic fungi, The count of UFC was done by the method of counting of fungi
mainly Colletotrichum gloesporioides. For this reason in the last 14 and yeasts in food based on the Ofcial Mexican norm NMX-F-255-
years, growers and packers are developing grove management 19788, which consists in prepare 100 mL of a sterile solution of 10%
techniques, packing practices and shipping practices in order to ex- tartaric acid, which was sterilized in autoclave for 15 min. at
port their avocados (Roberts and Orden, 1996) with attractive re- 121 C, cooled at 4548 C and to acidied at pH 3.5. Dilutions of
sults. The short shelf life of Mexican avocados is the biggest 1:1000, of avocado pulp in sterile tartaric acid solution were used.
problem to solve. 1 mL of dilution was added and distributed on petri plates with po-
In this work, the effort was centered in the elaboration of edible tato dextrose agar, Two sets of three petri plates for sample were
lm with ellargic acid at three different concentrations and to eval- prepared; one set was incubated at 22 C for 5 days and another
uate its effect in the physiochemical and microbiological quality of set at 35 C for 48 h.
whole avocados.
2.4. Experimental design and data analysis

2. Materials and methods Data were analyzed using a complete randomized block design,
with three replications. ANOVA and a test of Tukey for means com-
2.1. Materials parison were used to analyze the results.

The wax used in this study was extracted from the plant
Euphorbia antysyphilitica, better known by its common name in 3. Results and discussion
the Mexican desert region, as candelilla using the commercial pro-
cess applied by the company CENEMEXTM. Dispersions of ellagic The lightness or L values of the samples were improved by the
acid (ICN reagents, Monterrey, Nuevo Leon, Mexico) at a concentra- edible lms (Fig. 1). The lightness of WOC samples were sharply
tion of 0.01% were prepared in a edible lm formulated by Saucedo damaged (P < 0.05) during the period of study; the best results
Pompa (2007). were obtained with ECEA. Ellagic acid antioxidant delays the
changes in brightness in greater proportion than fruits WOC, The
2.2. Shelf life assays formulations with the active compound reduce the changes of col-
or by browning, including the control wax (ECC) as compared with
Hass avocados (Persea americana) were obtained from a local samples changes in appearance without wax (WOC). Parameter
supermarket in Saltillo, Mxico. The fruits were selected taking L related to luminosity is an excellent index for evaluation of color
into account their size maturity, visual absence of microorganisms in fruits (Saucedo-Pompa et al., 2007). Maia-Geraldine, Ferreira-
and damage on the skin. The fruits were later disinfected with a Soares, Alvarenga-Botrel, and de Almeida-Gonalves (2008) re-
solution of sodium hypochlorite (0.2 g/L) for 5 min, and dried at ported that the implement of edibles lms based on agar, chitosan
room temperature (Gonzales-Aguilar, Monroy-Garcinia, Goycoo- and acetic acid in garlic achieving lower the increase in color, while
lea-Valencia, Diaz-Cinco, & Ayala-Zavala, 2005). Tawil Bouchez (2003) reported that the use of a chitosan coating in
Edible lms were prepared in three formulations named as fol- apples reduced the color change. In another study with tomatoes
lows: ECEA, edible coating with ellagic acid; ECC, edible coating covered with an edible lm, the authors noted that after 22 days
without active compound (control lm). Negative controls were of storage, control fruits matured faster than the fruits with deck
fruits without edible lm (WOC). (Ghaouth, Ponnampalam, Castaigne, & Arul, 1992).
Each avocado was immersed for 1 s into a corresponding formu- Ruz-Cruz (2002) demonstrated the positive effect of adding dif-
lation (density: 0.86 g/mL). Each fruit was covered with 0.5 g of ferent antioxidants (independent or mixed) on the inhibition of
edible lm. The avocados were weighted before with an analytical browning of fresh-cut pineapple. When antioxidants are used in
 S. Saucedo-Pompa et al. / Food Research International 42 (2009) 511515 513

35 25

30
20
25

Lost of weight (%)

15
20
L vaue

15
10

10
5
5

0 0
0 1 2 3 4 5 6 7 0 1 2 3 4 5 6 7
Time (weeks) Time (weeks)

Fig. 1. Effect of edible lms on color (L value) of avocados with ECC (), ECEA (j) Fig. 3. Effect of edible lms on the lost of weight of avocados with ECC (), ECEA
and WOC (N). (j) and WOC (N).

combination with process technologies such as heat treatments, control the reduction in weight (Gonzlez-Aguilar et al., 2005).
modied and controlled atmospheres, edible lms, gamma radia- These results are similar to those found by Kester and Fennema
tions and electric pulses, the browning of fruits is inhibited. This in 1986 who report that the use of lipid-based coating retard mois-
study is the rst report on the use of a potent nutraceutical agent, ture loss, just as Assis and Pessoa (2004) reported that these coat-
ellagic acid as antibrowning additives in fruits, showing excellent ings applied to apples reduce the loss of moisture. Guilbert (1988)
results. used casein and carnauba wax in a coating formulation, this edible
All fruits presented changes in their external and internal visual covers reduce the loss of moisture in papaya, this is due to the bar-
appearances, but the avocados with ECEA suffered a minor change rier that is formed around the fruit, which prevents the loss of
compared with other treatments, especially compared with the water vapor (Ghaouth et al., 1991).
WOC avocados (Fig. 2) despite the extreme conditions of contam- Solids content in avocados with ECEA was higher in comparison
ination. ECC samples suffered a severe damage, but it was lower with the control treatments. May be, this is related to the acceler-
than that of WOC samples. Polysaccharide lms have been used ated maturation of the control fruits and the corresponding weight
to prevent changes in sliced fruits like lychee, tomatoes, papaya loss in the same (Fig. 4) (P < 0.05); in this case, volatile and water
and apples (Buta, Moline, Spaulding, & Wang, 1999; Gonzlez-Agu- soluble compounds can be released during the dehydration of con-
ilar et al., 2005). Similar results can be obtained with the new lms trol samples. With WOC and ECC treatments, the mass transfer in
formulated and applied in this study. And Ghaouth, Arul, and Ponn- the surface of fruits is the predominant factor of respiration (Rog-
ampalam (1991) reported that the use of edible coatings making ers, 1985).
slow changes in apple appearance, they argue that it is, due to Avocados coated with ECEA (P < 0.05), retained the initial rm-
the modied atmosphere created in the domestic fruit, with high ness values. The rmness of WOC samples was severely affected in
levels of CO2 and low O2 which delayed the maturation processes. comparison with those stored with lms containing antioxidants
The benets of using ellagic acid with candelilla wax as edible lm The texture of avocados was improved with the application of edi-
have been clearly evidenced by Saucedo-Pompa et al. (2007) due ble lms similar to the results reported by Ghaouth et al. (1992) for
they demonstrated that these lms improve the color and texture tomatoes.
of fruits. The pH values increased during the period of study for all trea-
In all samples, the weight loss was increased during the period ted samples (Fig. 5). However, this change in pH was signicantly
of study (P < 0.05), (Fig. 3). The highest lost of weight was found in low in comparison with WOC samples, while better results were
WOC samples. The lowest lost of weight was registered in samples obtained with ECEA (P < 0.05).
with ECEA. The use of ellagic acid as part of edible lms diminished These results are consistent with those obtained by Gonzlez-
the moisture loss in fruits. For fruits, the reduction in weight loss is Aguilar et al. (2005) they found a decrease in the increase of pH
very important and the use of edible lms is an excellent tool to of papaya treated with covers. On the other hand, Cantwell and

Fig. 2. Internal appearance of avocados with ECEA (a), ECC (b) and WOC (c).
514 S. Saucedo-Pompa et al. / Food Research International 42 (2009) 511515

70

65

60
Content of solids (%)

55

50

45

40

35

30
0 1 2 3 4 5 6 7
Time (weeks)

Fig. 4. Content of solids of avocados with ECC (), ECEA (j) and WOC (N).
Fig. 6. aw of avocados with ECC (), ECEA (j) and WOC (N).

Portela (1997) observed an increase in the pH of melon stored in an

atmosphere of air at 5 C.
Table 1
Avocados were signicantly beneted with edible lms con- Maximal values of the counting of colony forming unit.
taining ECEA (Fig. 6) (P < 0.05). The increment of aw values in
Treatments CFU per gram of avocado pulp
WOC samples because of the accumulation of water was outstand-
ing. The ECC and ECEA samples also showed the positive effect of Maximum load Incubation Time (h)
candelilla wax alone on the aw values. ECEA 40 240
The application of the edible lms on the external part (cracked) ECC 67 192
(P < 0.05) of the fruit demonstrated to be an effective barrier WOC 3373 240

against the penetration from the fungal contamination to the inter-

nal part of the fruit (pulp). ECEA maintains the quality of shelf life
what transforms it into the best option of avocados in fresh, all this 4. Conclusions
taking into account the extreme conditions of storage in those that
work (Table 1). These results are similar to those found by Maia- Application of edible lms based on candelilla wax and ellagic
Geraldine et al. (2008) who reported that the use of agar-based acid signicantly minimized the changes in appearance, solid con-
coatings, in garlic delayed 50% the fungal contamination in this tent, pH, aw, luminosity and weight loss, maintaining the quality of
food. avocado fruits and prolonging their shelf life compared with other
treatments, this without even leaving aside that even the ECC sig-
nicantly improves the quality of the shelf life of fruits, that be-
cause of the atmosphere which creates the application of this EF.
7.1 These excellent results were obtained considering the extreme
conditions of fungal contamination sued in this work. Finally, we
conclude that the use of an edible lm based on candelilla wax
7
with a potent antioxidant ellagic acid on whole fruits is a good
alternative to conserve fresh avocados.
6.9

6.8 Acknowledgements

The authors are grateful to the Mexican Program of CONACYT-

6.7
pH

CONAFOR (Project No. 2008-C01) for the nancial support of this

work. Authors Saul Saucedo-Pompa and Romeo Rojas Molina want
6.6
to thank to Mexican Council of Science and Technology (CONACYT)
by their scholarships received for their postgraduate studies.
6.5

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