Industrial Crops and Products 60 (2014) 3944

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Industrial Crops and Products

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Antioxidant and antimicrobial activities of Psidium guajava L. spray

dried extracts
M.R.V. Fernandes a , A.L.T. Dias b , R.R. Carvalho b , C.R.F. Souza a , W.P. Oliveira a,
a
b

Faculdade de Cincias Farmacuticas de Ribeiro Preto, Universidade de So Paulo, USP, Av. do Caf s/n, Ribeiro Preto, SP, 14040-903, Brazil
Universidade Federal de Alfenas, Instituto de Cincias Biomdicas. Rua Gabriel Monteiro da Silva, 700, Alfenas, MG, 37130-000, Brazil

a r t i c l e

i n f o

Article history:
Received 24 January 2014
Received in revised form 8 May 2014
Accepted 31 May 2014
Keywords:
Spray drying
Psidium guajava
Antimicrobial activity
Antioxidant activity
Pearson correlation

a b s t r a c t
Psidium guajava L. (guava) is widely used in traditional medicine for the treatment of diarrhea, gastroenteritis and as anti-inammatory, among other uses. This research evaluated the antimicrobial
and antioxidant activities of spray dried extracts from leaves of Psidium guajava L. (SDGEs). Different
technological adjuvants, namely: maltodextrin (MD), colloidal silicon dioxide (A), arabic gum (E) and cyclodextrin (CD) at concentration of 8% (wet base) were added to drying composition. SDGEs were
characterized through determination of the total phenolic and avonoid content. The extracts were
tested against fungi (Candida albicans, Candida krusei and Candida glabrata), Gram-positive bacteria
(Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa) by
the microdilution assay. The antioxidant activity of the SDGEs was assessed by the DPPH protocol. The
results showed that the MDEA and CD SDGEs were more effective against C. glabrata (MIC = 80 g/mL),
followed by MA SDGE which exhibited activity against C. krusei and C. glabrata also (MIC = 100 g/mL).
The better inhibitory effect on S. aureus was observed to the MA SDGE with a value of 100 g/mL. The
IC50 of the SDGEs ranged from 7.96 to 8.11 g/mL by DPPH method. The results indicates signicant
antimicrobial and antioxidant activities of Psidium guajava SDGEs presenting high potential as an active
phytopharmaceutical ingredient for development of herbal medicines.
2014 Elsevier B.V. All rights reserved.

1. Introduction
It is well-known that plant-derived compounds are potential sources of new antibiotics, anticancer, and anti-HIV drugs
among others pharmaceutical compounds (Gurib-Fakim et al.,
2005). Important groups of phytochemicals are alkaloids, tannins,
avonoids and phenolic compounds (Amal et al., 2009). Flavonoids
are the largest and best studied group of polyphenols. These compounds are a class of secondary plant metabolites that are thought
to exert benecial health effects through their antioxidant and
chelating properties which are the major contributor to the antioxidant capacity (Williams et al., 2004). In addition to the antioxidant
activities, many phenolic compounds have been shown to exhibit
antimicrobial actions (Metwally et al., 2011; Prabu et al., 2006).
The study of antimicrobial and antioxidant substances from natural sources has received increased interest; and Brazil possess a
huge biodiversity and a signicant number of plants have been used
in folk medicine for hundreds of years (Silva et al., 2010).

Corresponding author. Tel.: +55 16 36024185.

E-mail addresses: wpoliv@fcfrp.usp.br, wpoliv@gmail.com (W.P. Oliveira).
http://dx.doi.org/10.1016/j.indcrop.2014.05.049
0926-6690/ 2014 Elsevier B.V. All rights reserved.

Generally, standardized plant preparations are marketed in

the form of liquid extracts, or as powders resulting from the
drying of plant material or by drying an extract (Souza et al.,
2008). In this context, the trend in the pharmaceutical industry is to use dried extracts since products in solid form present
signicant advantages over conventional liquid forms, such as
chemical, physicalchemical and microbiological stability; ease of
standardization; higher concentration of active compounds; ease of
transportation; less space for product storage and less risk of microbial contamination. Transforming a plant extract into dry extract is
also widely used in developing herbal medicinal products (Oliveira
et al., 2010). Among the drying technologies used in the preparation of dried extracts, spray drying predominates. Attention should
also be addressed to the processing conditions in order to preserve
the bioactive compounds linked to its pharmacological activities
(Gorinstein et al., 2009).
Spray drying has been used by several authors to obtain active
phytopharmaceutical ingredients and thereby guaranteeing the
efcacy, quality and safety of the product (Bott et al., 2010; CortsRojas and Oliveira, 2012; Kha et al., 2010; Souza et al., 2008).
Various technological adjuvants, including maltodextrin, modied
starch, cyclodextrins, arabic gum and colloidal silicon dioxide, are

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M.R.V. Fernandes et al. / Industrial Crops and Products 60 (2014) 3944

generally added to extractive solutions before spray drying in order

to improve process performance and product quality (Oliveira et al.,
2012).
Psidium guajava (Myrtaceae), popularly known as guava, is originally from Central and South America and is cultivated in all tropical
and subtropical countries (Alves et al., 2006). Extracts and metabolites of this species, especially obtained from the leaves, possess
useful pharmacological activities including antioxidant and antimicrobial properties, and have been widely used especially in the
treatment of diarrhea. Other reported uses include its function as
a hypoglycemic, antitussive, anti-inammatory and antioxidant,
thereby reinforcing its traditional use (Gutirrez et al., 2008). In
Brazil, it is inserted in the list of plant species selected for study by
the Research Program on Medicinal Plants (Programa de Pesquisas
de Plantas Medicinais, PPPM) and included in the National Report of
Medicinal Plants of Interest to the Public Health System (RENISUS,
2012).
The objective of this research was to evaluate the antioxidant
and antimicrobial activities of spray dried extracts of Psidium guajava leaves. The search of natural antimicrobial agents is highly
justiable due to problem of antimicrobial resistance observed with
current conventional treatment.
2. Materials and methods
2.1. Chemicals and reagents
Ethanol absolute, aluminum chloride, sodium tungstate and
phosphomolybdic acid (Vetec Qumica Fina Ltda, Duque de Caxias, RJ - Brazil); gallic acid, 1,1-diphenyl-2-picrylhydrazyl (DPPH;
SigmaAldrich, St. Louis, MO - USA), dimethylsulfoxide (DMSO),
and Mueller Hinton broth (Merck, Schuchardt, Hohenbrunn
Germany) were the main chemicals used in this study. The technological adjuvants used were: colloidal silicon dioxide (Aerosil 200;
Evonik Degussa, Hanau, Germany), maltodextrin (MOR-REX 1910;
Ingredion Brasil, Mogi Guacu, SP - Brasil), -cyclodextrin (Kleptose; Roquette, Lestrem FR), and gum Arabic (Encapsia; NEXIRA
do Brasil, So Paulo, SP, Brazil).
2.2. Plant materials
The leaves of Psidium guajava were collected in 2011 from the
Casa da Goiaba farm and industry (Lavras MG, Brazil), and taxonomically identied by Prof. Dr. Marcelo Polo of the Universidade
Federal de Alfenas (UNIFAL-MG, Brazil). Voucher specimens were
deposited at the herbarium of the UNIFAL, being cataloged with the
number UALF-01505.

Table 1
Technological adjuvants and codes to spray dried extracts from leaves of Psidium
guajava.
Samples

Technological adjuvants

Adjuvant ratio (%)

MA
MDEA
CD

Maltodextrin DE10: Aerosil

Maltodextrin DE10: Encapsia : Aerosil
-cyclodextrin

7:1
5:2:1
8

2.4. Spray drying

Drying was performed in a bench-top Lab-Plant SD-05 spray
dryer (Lab-Plant UK Ltd., Hudderseld, UK) with a concurrent ow
regime. The drying chamber had an internal diameter of 215 mm
and a height of 500 mm. The drying air was supplied by a blower
connected to an air lter. The temperature was controlled by a
temperature controller (included in the setup). The herbal extract
preparations were fed into the dryer by a peristaltic pump connected to a two-uid atomizer, with an internal orice of 0.7 mm.
The product was collected in a Lapple cyclone with a column diameter of 0.085 m (cut diameter of 3.9 mm). The operating parameters
were set according to previous studies carried out by our research
group (Corts-Rojas et al., 2012; Georgetti et al., 2008; Souza and
Oliveira, 2006): inlet air drying temperature, Tgi = 150 C; drying
air ow rate, Wg = 60 m3 /h; extract feed ow rate, Wsusp = 4 g/min;
atomizing air pressure, Patm = 1.5 bar and atomizing air ow rate,
Watm = 15 lpm. During the drying experiments, the room temperature and relative humidity (%) were monitored by using a digital
hygrometer (Minipa MTH model, 1361).
The technological adjuvants were added to the concentrated
extract at proportion of 8% (wet base) before spray drying runs.
The adjuvants proportions and codes of the spray dried extracts
(SDGEs) are presented in Table 1.
2.5. Chemical characterization of the extractive solution,
concentrated extract and dried extracts
2.5.1. Total phenols determination
Total polyphenol (TP) contents were quantied using
the Folin-Denis method, which involves the reduction of
phosphomolybdicphosphotungstic acid by phenolics in alkaline medium resulting in an intense blue color that is measured
in a UV/vis HP 8453 spectrophotometer (Agilent Technologies,
Waldbronn Germany) at wavelength of 750 nm after a reaction
time of 2 min (Folin and Denis, 1912; Souza et al., 2008). The
results are expressed in terms of gallic acid equivalents per gram of
extract or gram of plant material using an analytical curve which
was prepared using 535 mg/L solutions of gallic acid in puried
water with a correlation coefcient of 0.9962. The samples were
analyzed in triplicate.

2.3. Extraction of plant material

The fresh leaves were dried in an oven drying at temperature
of 45 C until constant weight, and ground in a knife mill (MA
mod. 680, Marconi Equipamentos Para Laboratrios Ltda, Piracicaba, SP Brazil), using a 20-mesh sieve (833 m). The powdered
material was maintained at room temperature in hermetic plastic
containers and protected from light until required. Extracts were
prepared through dynamic maceration under constant stirring for
60 min, using ethanol:water 70% (v/v) at 50 C. The plant/solvent
ratio was set at 1:10 (w/v). The extractive solution (ES) obtained
was ltered and the solvent was removed in a rotary evaporator
(Fisatom, mod. 802) at 50 C under vacuum of 650 mmHg, until
solid contents reached 11.8% (w/w). The concentrated guava extract
(CGE) was spray dried in presence of different technological adjuvants. The SDGEs and CGE were analyzed for total phenolic and
avonoid contents and antioxidant activity by DPPH assay.

2.5.2. Total avonoids determination

The total avonoid contents (TF) was quantied using a
spectrophotometric method based on the displacement of the
wavelength at 425 nm following the addition of 0.5% AlCl3 (w/v)
with a reaction time of 30 min (Souza, 2007). Total avonoid
contents was expressed in milligrams of quercetin per gram of dried
product, as determined by an analytical curve which was prepared
by preparing quercetin solutions at 120 mg/mL concentrations in
ethanol 40% with a correlation coefcient of 0.9973. The samples
were analyzed in triplicate.
2.6. Determination of antioxidant activity by DPPH radical assay
DPPH is a stable free radical that reacts with compounds, which
are able to donate a hydrogen atom. Thus, the hydrogen donating
ability of the SDGEs and CGE to DPPH was determined from the

M.R.V. Fernandes et al. / Industrial Crops and Products 60 (2014) 3944

41

Table 2
IC50 values for radical scavenging activity, radical scavenging capacity (% Inhibition), total avonoids and total phenolic of the concentrated guava extract, extractive solution
and spray dried guava extracts of Psidium guajava.
Sample

IC50 (g mL1 )a

Inhibition (%)

TF (mg/g, db)

TP (% m/m, db)

MA 80
MDEA 80
CD 80
CGE
ES
Quercetin

8.11 (0.05)*
9.76 (0.21)*
7.96 (0.18)*
3.34 (0.11)*
3.94 (0.10)
0.96 (0.01)

88.54 (0.51)
85.89 (0.90)
86.09 (0.84)
88.48 (0.28)
82.92 (1.36)
86.20 (0.90)

13.35 (0.32)
15.48 (0.87)
12.58 (0.17)
22.58 (0.08)
23.48 (0.20)

10.52 (0.35)
9.67 (0.1)
12.47 (0.66)
23.21 (0.72)
25.93 (0.80)

a
Antioxidant activity by the DPPH method, expressed as IC50 ; TF: total avonoids; TP: total polyphenols; db: dry base.
p < 0.05. *: Statistical difference determined by Dunns post-test. CGE: concentrated guava extract; ES: extractive solution.
Values are presented as the mean (standard deviation).

change in absorbance at 517 nm, according to Blois (1958) method.

For radical scavenging measurements, 1 mL of 0.1 M acetate buffer,
pH 5.5, 1 mL of ethanol, 0.5 mL of 250 M ethanolic solution of
DPPH were mixed, 10 L of each extract under study were added,
then the absorbance was measured after 20 min at room temperature. The blank was prepared from the reaction mixture without
DPPH solution. Quercetin was used as the reference sample. The
results were expressed as IC50 (g mL1 ), which means the amount
of sample necessary to decrease the absorbance of DPPH in 50%. All
measurements were performed in triplicate (Georgetti et al., 2006;
Souza et al., 2009).

Dunns post-test to reveal statistical differences between groups

(Hollander and Wolfe, 1999; Pagano and Gauvreau, 2004), using a
signicance level of 5%. The results were obtained with the aid of
the software R 2.14.0, using the PGIRMESS library of functions (R
Development Core Team, 2011).
For the analysis of correlations between the antioxidant activity by DPPH method and polyphenols content was determined the
Pearson correlation coefcient (r) using the SAS software version
9.0 (SAS/STAT Users Guide, 2002).

2.7. Antibacterial and antifungal activity evaluation

3.1. Determination of antioxidant activity

The CGE, the MA, MDEA and CD SDGEs and the technological
adjuvants maltodextrin, Aerosil , arabic gum and -cyclodextrin
were evaluated in vitro for their antimicrobial activities against
the fungi through a Mueller Hinton broth microdilution method
and with the methodology and interpretative criteria proposed by
document M27A3 (CLSI, 2008) and through a standard Mueller
Hinton broth microdilution method for bacteria proposed by document M7A6 (CLSI, 2003). The standard pathogenic/opportunistic
fungi were Candida albicans (ATCC 10231), Candida krusei (ATCC
6258), Candida glabrata (ATCC 90030) and bacteria, the Gram positive Staphylococcus aureus (ATCC 6538) and the Gram negative
Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC
9027). The stock solutions of all the compounds were prepared in
DMSO 1% at nal concentration and tested at concentrations from
1000 to 5 g mL1 by serial dilution. The standard drug uconazole was applied as control of fungistatic action at concentration
from 64 to 0.031 g mL1 and the standard drug chloramphenicol
was applied as a control of bacteriostatic action at concentrations from 125 to 0.06 g mL1 . The microplates were incubated
at 35 C for 24 h for bacteria and 37 C and for 24 h for fungi. Results
were visualized and analyzed at 530 nm in an Anthos Zenyth 200
rt Microplate Reader. The minimal inhibitory concentration of
microbial growth was determined at 50% (MIC50 ) in g mL1 and
compared for each compound and microorganism. The minimal
bactericidal and fungicidal concentrations (MBC and MFC, respectively) were also determined. The MBC was assessed by plating
the volume of 20 L from the broth microdilution MIC well for
each microorganism onto Mueller Hinton Agar (for bacteria) and
Sabouraud Agar (for fungi). After incubation at 37 C for 24 h, the
least concentration showing no visible growth on sub-culture was
taken as the MBC or MFC. The tests were all done in duplicates and
the results obtained from the replicas were coincident.
2.8. Statistical analysis
The statistical analysis of the experimental data was carried
out with the aid of SAS software, version 9.0 (SAS/STAT Users
Guide, 2002). The Kruskall Wallis test was also used, followed by

3. Results and discussion

DPPH is a free radical, stable at room temperature, which

produces a violet solution in ethanol. The reduction of DPPH is
monitored by decrease of the absorbance of its radical at 517 nm.
It is reduced in the presence of an antioxidant molecule resulting in uncolored ethanol solutions. The DPPH assay is an easy and
rapid way to evaluate antioxidant activity (Silva et al., 2005). Results
of DPPH reduction produced by all extracts tested are shown in
Table 2. The potential antioxidant of the CGE showed a signicant
free radical scavenging activity comparing with the high antioxidant effect of quercetin (a potent natural antioxidant). With respect
to IC50 , DPPH radical scavenging activities of SDGEs varied from
7.96 to 9.76 (g mL1 ), which represents an antioxidant potential
2.53 times lower than the concentrated guava extract (Table 2).
It is well-known that the lower IC50 , the higher is the antiradical
efciency (Magalhes et al., 2008). However, the SDGEs showed
the lowest antioxidant potential due only to the dilution effect
caused by incorporation of technological adjuvants. Both IC50 and
% Inhibition values were calculated on the basis of the solids content of extractive solution. The total phenolic contents of the CGE
and SDGEs are also shown in Table 2. The total avonoid content of
the SDGEs in terms of quercetin equivalent was between 12.58 and
15.48 (mg/g, dry base). The total phenolic content of SDGEs ranged
from 9.67 to 12.47 (% m/m, dry base). The difference in phenolic
compounds content between SDGEs and CGE was not statistically
signicant, since the values were also calculated on the basis of
the solids content of concentrated extract. In addition, no significant degradation of phenolic compounds in relation to MA and
MDEA SDGEs was observed. The CD SDGE presented a small rate
of degradation compared to the total avonoid content (3%). The
degradation during drying is dened as the ratio of the difference
between the compound (avonoid or phenolic) concentration in
the concentrated extract and dried extract relative to concentrated
extract (Bott et al., 2010).
There are several studies showing the antioxidant activity of
guava leaves extracts which included important review articles
(Barbalho et al., 2012; Gutirrez et al., 2008). Nonetheless, information concerning the antioxidant activity of spray dried guava
leaves extracts is unavailable.

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M.R.V. Fernandes et al. / Industrial Crops and Products 60 (2014) 3944

Ayoola et al. (2008) used the DPPH method to determine the

antioxidant activities of ethanolic extracts from leaves of Carica
papaya L., stem bark of Magnifera indica L., leaves of Psidium guajava L. and the leaves of Vernonia amygdalina Del., medicinal plants
traditionally used in the treatment of malaria in southwestern
Nigeria. The concentration of the P. guajava extract required for
50% inhibition of DPPH radical scavenging (IC50 ) was 40 g mL1 .
The crude extract of P. guajava appeared to be as potent as vitamin
C with a maximum inhibition of 91% at 500 g mL1 . The results of
antioxidant activity obtained in the present study evidenced a signicant higher free radical scavenging in the DPPH assay (Table 2).
It is noteworthy to mention that the mechanism involved in the
reduction of DPPH free radicals is based on a scavenging activity. Thus, the structure (both planar and spatial) of the antioxidant
compound, present in the extract, is important for its capacity of
donating hydrogen ions (Tian and McLaughlin, 2000). Moreover,
the main limitation of IC50 determination is that the percentage
of radical scavenged is dependent on the initial concentration of
DPPH radical (Magalhes et al., 2008). Consequently, the antioxidant activity measured by the DPPH method can present signicant
differences between the results of the assays. In addition, production of plant secondary metabolites depend on a number of intrinsic
(genus, species, cultivars) and extrinsic (agronomic, environmental, handling and storage) factors (Kalt et al., 2001; Toms-Barbern
and Espn, 2001), which can lead to signicant variation between
the phenolics contents of herbal materials from different sources
(Balasundram et al., 2006).
The free radical scavenging effects of aqueous extracts from
leaves of Psidium guajava L., Camellia sinensis (GABA tea), Toona
sinensis Roem. and Rosmarinus ofcinalis L. were reported by Chen
et al. (2007). Among the four extracts examined, the Psidium
guajava exhibited the strongest efciency and showed over 50%
scavenging effect of DPPH at a concentration of 100 g mL1 ,
evidencing the highest hydrogen-donating capacity; although signicant lower than the values determined in this work. Qian and
Nihorimbere (2004) found that the ethanol guava leaves extracts
showed an antioxidant activity expressed as percentage of inhibition of 82.7% at 500 g mL1 concentration. The different extracts
from guava leaf showed good free radical-scavenging activity
depending on the concentration used. The higher the concentration
used the higher the free radical-scavenging effect (dose dependent).
The radical scavenging activity of methanolic and aqueous
extracts Psidium guajava leaves was also investigated by the DPPH
method by Venkatachalam et al. (2012). It was found that the
methanolic extracts showed greater amount of phytochemicals
and higher antioxidant activity (42 mg ascorbic acid equivalent)
than aqueous extracts (39 mg ascorbic acid equivalent in aqueous
extract). According to the authors, guava leaf extracts are a rich
source of natural antioxidants and good candidate for development
functional foods or herbal remedies with chemopreventive effects.
3.1.1. Correlation between total phenolic/avonoid content and
antioxidant activity
It is known that the polyphenols are effective antioxidants due
to their ability to act as hydrogen donors, reducing agents and radical scavengers (Mai et al., 2009). The leaves of guava are rich in
avonoids and phenols which contribute to its antioxidant potential (Chen and Yen, 2007).
The correlations between polyphenols content (TP) and
avonoids content (FT) with antioxidant activity measured by
DPPH assay were also evaluated with the Pearson correlation coefcient. The results showed a strong correlation between DPPH
scavenging activity (IC50 ) and phenolics content (r = 0.980) and
avonoid content (r = 0.976). The antioxidant activity measured
by the DPPH method (IC50 ) showed strong negative correlation

Table 3
Minimum inhibitory concentration (MIC) and minimum bactericidal concentration
(MBC) of spray dried Psidium guajava extracts and concentrated guava extract.
Sample

MA
MDEA
CD
CGE
Adjuvants**
Chloramphenicol

MIC (MBC) g/mL

S. aureus

E. coli

P. aeruginosa

100 (*)
250 (500)
125 (1000)
100 (1000)

0.975

1000 (*)
1000 (*)
250 (*)

0.975

1000 (*)
1000 (*)
1000 (*)
500 (*)

31.2

CGE: concentrated guava extract

**: Adjuvants: maltodextrin, Aerosil , arabic gum and -cyclodextrin. The substance
did not show growth inhibitory activity of the microorganism in the concentration
evaluated;
Parentheses value represents minimum bactericidal concentration (MBC).
(*): there were not MBC values at the concentration ranges evaluated.

with TF and TP. This indicates that the lower the IC50 value, the
higher the content of FT and PT. Youwei et al. (2008) reported
excellent correlation between polyphenolic content and free radicals in fresh owers when antioxidant activity [DPPH (Free Radical
Scavenging Activity: FRSA)] was compared with total polyphenolic
(r = 0.983, p < 0.01). The data obtained leads to the conclusion that
polyphenolic compounds could be taken into account for the strong
FRSA, which are in agreement with others studies. Silva et al. (2007)
also suggested that polyphenolic content could be related to the
antioxidant activities.
Therefore, the results of this study suggest that phenolic compounds are a major contributor to antioxidant capacity in the guava
leaf extracts.
3.2. Antimicrobial activity
The life-threatening diseases like bacterial ones and mycoses
caused by opportunistic bacterial (mainly S. aureus, P. aeruginosa)
and fungal pathogens (mainly species from the genus Candida)
associated with nosocomial infections, are one of the major health
problems in present days. Therefore, it is crucial to identify new
compounds, especially natural ones, which are active against the
most, broad spectrum of Gram positive, Gram negative and Candida
species.
The antibacterial activity of CGE and SDGEs (and adjuvants) of
Psidium guajava against Gram positive and Gram negative bacteria
by microdilution method is shown in Table 3.
The technological adjuvants used in spray drying were also evaluated separately in the same concentrations of the extracts and it
was veried they did not present growth inhibitory activity of the
tested microorganisms (both bacteria and fungi).
According to Holetz et al. (2002), extracts that showed values
of MIC less than 100 g mL1 , the antimicrobial activity was good;
from 100 to 500 g mL1 the antimicrobial activity was moderate; and from 500 to 1000 g mL1 the antimicrobial activity was
weak; over 1000 g mL1 the extract was considered inactive. It
was evaluated the antimicrobial activity of hydroalcoholic extracts
of 13 plants against Gram positive bacteria, Gram negative bacteria
and some yeast. The Psidium guajava extract was also evaluated presenting a moderate activity against S. aureus (MIC = 250 g mL1 )
and E. coli (MIC = 500 g mL1 ). These results are in close agreement
with the ones presented in this work, since the most relevant results
were against S. aureus which ranged from 100 to 250 g mL1 .
Sanches et al. (2005) showed results similar to the ones observed
in the present study. The authors assessed the activity of waterethanol extracts (70:30 v/v) from leaves of P. guajava against S.
aureus (MIC value of 125 g mL1 and MBC value of 250 g mL1 )
by the broth microdilution method. Moreover, the extract was

M.R.V. Fernandes et al. / Industrial Crops and Products 60 (2014) 3944

Table 4
Minimum inhibitory concentration (MIC) and Minimum fungicidal concentration
(MFC) of spray dried Psidium guajava extracts.
Sample

MA
MDEA
CD
CGE
Adjuvants**
Fluconazole

MIC (MFC) g/mL

C. albicans

C. glabrata

C. krusei

1000 (*)
250 (500)

1 (*)

100 (1000)
80 (*)
80 (1000)
80 (250)

8 (*)

100 (*)

1000 (*)
100 (250)

32 (*)

CGE: concentrated guava extract;

**: Adjuvants: maltodextrin, Aerosil , arabic gum and -cyclodextrin. The substance
did not show growth inhibitory activity of the microorganism in the concentration
evaluated;
Parentheses value represents minimum fungicidal concentration (MFC).
(*): there were not MFC values at the concentration ranges evaluated.

inactive against Gram negative bacteria E. coli and P. aeruginosa

(MICs > 1000 g mL1 ).
On the other hand, Betoni et al. (2006) evaluated watermethanol extracts of P. guajava leaves against S. aureus displaying
MIC value of 560 g mL1 . In our study, the CGE showed a moderate
activity against E. coli (MIC value of 250 g mL1 ) and P. aeruginosa
(MIC value of 500 g mL1 ), and the SDGEs did not exhibit activity against Gram negative bacteria. This can be awaited since the
outer membrane of Gram negative bacteria present a barrier to
the penetration of numerous antibiotic molecules, as well as their
periplasmic space contains enzymes, which are able to inactivate
molecules from outside the bacterial cell, for instance, antibiotics
(Duffy and Power, 2001).
Table 4 shows the MICs and MFC of the SDGEs, CGE and the
technological adjuvants tested against three Candida species.
According to the criteria adopted by Holetz et al. (2002), all
of SDGEs evaluated had good antimicrobial activity against the
C. glabrata, although did not exhibit activity against C. albicans.
The CGE and the SDGEs presented their better antimicrobial activities against C. glabrata, showing in vitro inhibitory potential to
treat infections by this yeast. C. glabrata is considered a common
commensal in gastrointestinal and genitourinary tracts, but can
turn into an opportunistic fungal pathogen in immunocompromised patients. This species possesses both innate and acquired
resistance against antifungal drugs, due to its ability to modify ergosterol biosynthesis, mitochondrial function, or antifungal
efux. This resistance allows for its relative overgrowth over other
susceptible species and may contribute to the recent emergence of
C. glabrata infections in chronically immunocompromised populations (Li et al., 2007). Holetz et al. (2002) evaluated the antimicrobial
activity of hydroalcoholic extract of P. guajava against yeasts (C.
albicans, C. krusei, C. parapsilosis and C. tropicalis) by the broth
microdilution method and showed that the extract had good activity against all tested yeasts (MIC = 15.6125 g mL1 ). Corts-Rojas
(2011) evaluated spray dried extracts from Bidens pilosa using technological adjuvants such as maltodextrin, arabic gum and Aerosil
against C. albicans, C. krusei, C. parapsilosis, C. tropicalis and C.
glabrata. All of these spray dried extracts did not exhibit antimicrobial activity against tested yeasts.
Interesting to note that, even though there was decrease of
antimicrobial activity due to the drying process and the dilution
effect caused by technological adjuvants, spray dried extracts have
greater chemical, physicochemical and microbiological stability.
This ensures in the long term, the antimicrobial activity of these
extracts.
In Brazil, the majority of the approved phytomedicines are formulated as solid dosage forms containing plant dried extracts as
active component. Spray drying technique has been widely used

43

to obtain dried extracts presenting better technological characteristics and greater concentration of biological active constituents.
Physicochemical properties of such products depend on factors
related to process, formulation (inlet material) and equipment
(Oliveira and Petrovick, 2010). According to our results, there was
a good correlation between the experimental data of antimicrobial
activity of spray dried preparations from Psidium guajava leaves
against the common human pathogens.
4. Conclusions
The results of the present study showed that the spray dried
guava leaves extracts present potential antioxidant and antimicrobial activities. Psidium guajava leaves extract showed antibacterial
activity against S. aureus and also showed antifungal activity against
C. glabrata. The spray dried guava leaves extracts exhibited stronger
antimicrobial activity against S. aureus and C. glabrata, evidencing their potential as a natural antimicrobial agent for medicinal,
cosmeceutical and food purposes. These results could be useful
for developing pharmaceuticals preparations for treating microbial
diseases, and should be further investigated in in vivo models.
In addition, it is important to observe that the spray drying
technique allows concentration of active compounds, since the concentrated extract showed antimicrobial and antioxidant activities.
Therefore, our results prompted us to conclude that spray dried
guava leaves extracts have been found to be promising sources of
potential antioxidant and antimicrobial agents.
Acknowledgements
The authors are grateful to the Brazilian Funding Agencies, CNPq
(Conselho Nacional de Desenvolvimento Cientco e Tecnolgico)
and FAPESP (Fundaco de Amparo a Pesquisa do Estado de So
Paulo) for the nancial support.
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