International Journal of Food Science and Technology 2010, 45, 403–409 403

Original article
Antimicrobial and antioxidant activity of crude onion
(Allium cepa, L.) extracts

1 2 3
Jonathan Santas, * Mar´ıa Pilar Almajano & Rosa Carbo´
1 Department of Nutrition and Food Science, University of Barcelona, Av. Joan XXIII, s ⁄ n, 08028, Barcelona, Spain
2 Chemistry Engineering Department, The Technical University of Catalonia, Av. Diagonal 649, 08034 Barcelona, Spain
3 Department of Agri-Food Engineering and Biotechnology, The Technical University of Catalonia, Campus Baix Llobregat, C. Esteve Terrades 8,
08860 Castelldefels (Barcelona), Spain
(Received 7 October 2009; Accepted in revised form 9 December 2009)

Summary Antioxidant and antimicrobial activity of the ethyl acetate and water subfractions of methanolic extracts of
three Spanish onion varieties were assayed. Flavonoids were mainly present in ethyl acetate subfraction
being 34.92 ± 0.75, 7.95 ± 0.16, 0.38 ± 0.01 lmol of rutin eq. g)1 D.W. and its antioxidant capacity
was
74.86 ± 1.77, 24.59 ± 0.67, 4.55 ± 0.44 lmol Trolox g)1 D.W. of Grano de Oro, Fuentes de Ebro
and
Calc¸ ot de Valls varieties, respectively. The antimicrobial activity of flavonol standards and onion extracts
was evaluated against some food spoiler microorganisms. Quercetin and kaempferol were inhibitory against
gram positive bacteria such as Bacillus cereus, Staphylococcus aureus, Microcroccus luteus and Listeria
monocytogenes. Gram negative bacteria Escherichia coli and Pseudomonas aeruginosa were less sensible to the
antimicrobial effect of both flavonol standards and Candida albicans was totally resistant. Among the onion
extracts tested only ethyl acetate subfraction showed antimicrobial inhibition.
Keywords Antimicrobial activity, antioxidant activity, flavonoids, onion.

Introduction
Since ancient times onions (Allium cepa, L.) have been
an important dietary resource and have also been of
interest for medical purposes (Rose et al., 2005). Tradi-
tionally onions and plants belonging to the Allium genus
have been used as an herbal remedy for a wide range of
ailments, due to their association with many pharma-
cological effects (Yin & Cheng, 1998; Rose et al., 2005).
Biological effects attributed to onions have been
commonly ascribed to the volatile sulfur-containing
compounds, such as thiosulfinates, mainly responsible
for the characteristic taste, aroma and lachrymatory
effects (Lanzotti, 2006). These compounds are formed
from cysteine sulfoxide precursors and the effect of the
enzyme alliinase which is released from cell vacuoles
when tissues are damaged (Krest & Keusgen, 2002).
However, these volatile products are highly unstable
and recently attention has been focused on the effects of
phenolic compounds, such as flavonoids, which are more
stable (Ioku et al., 2001). Onion is known for being a
good natural source of flavonoids mainly represented by
the flavonols quercetin and kaempferol, which are
present as their glycosides (Fossen et al., 1998).
*Correspondent: E-mail: jonathan.santas@ub.edu
In recent years, many publications have reported
evidence of beneficial health effects attributed to
flavonoids including antiallergenic, antiinflammatory,
cardioprotective, vasodilatory, anticarcenogenic and
antioxidant properties (Shon et al., 2004). Several epi-
demiological studies have also associated the consump-
tion of flavonoids with a reduction of the risk of chronic
diseases including, cancer, diabetes and coronary heart
problems (Hirvonen et al., 2001; Kosmider & Osiecka,
2004).
In addition, many flavonoids have been reported to
posses antibacterial and antifungal properties (Rauha
et al., 2000). This has increased the interest of the food
industry in these natural compounds as components to
improve food stability against microbiological spoiling
agents (Taguri et al., 2004; Sofia et al., 2007). Protection
of food from pathogens and spoilage organisms has
been traditionally achieved by chemical methods, but
during recent years there has been an increase in
consumer interest in developing foods which contain a
low level or are free of chemical preservatives (Viuda-
Martos et al., 2008). The emergence of pathogens which
are resistant to classical preservatives has also created an
urgent necessity to find alternatives as antimicrobial
agents (Xu & Lee, 2001). In consequence, the food
industry is interested in developing natural components

doi:10.1111/j.1365-2621.2009.02169.x
2010 The Authors. Journal compilation 2010 Institute of Food Science and Technology
40 Antimicrobial and antioxidant activity of A. cepa extracts J. Santas et al.
4 4
for the total or partial replacement of synthetic antimi-
crobials (Grohs & Kunz, 2000).
Onions can be considered as a good source of natural
additives to retard food deterioration (Navas et al.,
2006). However, the application of thiosulfinates and
volatile compounds for food preservation is limited due
to their strong flavour and their biochemical instability
(Benkeblia, 2004). These properties focus attention on
the more stable flavonoids as additives to enhance food
shelf-life by inhibiting microbial spoiling and oxidative
deterioration, due to their antimicrobial and antioxidant
properties (Ramos et al., 2006; Naz et al., 2008).
The aim of this research is: (1) to determine the total
phenol content and antioxidant capacity of crude onion
extracts, (2) to evaluate the antimicrobial activity of onion
extracts against the spoilage and pathogenic food-related
microorganisms: Bacillus cereus, Staphylococcus aureus,
Microcroccus luteus, Listeria monocytogenes, Escherichia
coli, Pseudomonas aeruginosa and Candida albicans and
(3) to compare the differences between hydrophilic and
hydrophobic subfractions from crude onion extracts.
Materials and methods
Plant material
Three different Spanish onion varieties (A. cepa, L.)
were purchased in a local market: white skinned onion
var. ‘Fuentes de Ebro’(FE), white skinned onion var.
‘Calc¸ ot de Valls’ (CV) and yellow skinned onion
var.
‘Grano de Oro’ (GO). Samples were stored in the dark at
)20 C until their analysis.
Onions were skinned, chopped, blended (Moulinex
Blender) and finally freeze-dried. The freeze-dried onions
were ground in a mortar until obtaining a fine powder and
were stored at 4 C in the dark and in a dry atmosphere.
Chemicals and standards
Methanol, ethyl acetate, sodium carbonate, dimethyl-
sulfoxide (DMSO), aluminium chloride, acetic acid and
Folin–Ciocalteu reagent were analytical grade from
Panreac (Barcelona, Spain). Water used for the study
was deioniser and pure water (Millipore-Q System).
Rutin, quercetin, kaempferol, 6-hydroxy-2,5,7,8-
tetramethylchroman-2-carboxylic acid (Trolox),
potassium persulfate and 2,2¢-azino-bis(3-ethylbenzo-
thiazoline-6-sulfonic acid) diammonium salt (ABTS)
were purchased from Sigma-Aldrich Company Ltd
(Gillingham, UK).
Preparation of the extracts
Light exposure was avoided during the extraction
process. Freeze-dried onion powder (3 g) was extracted
with 30 mL of 75% methanol. After 30 min of extrac-
International JournalJournal
2010 The Authors. of Food Science and
compilation 2010 Technology
Institute of Food Science and
2010
Technology
Antimicrobial and antioxidant activity of A. cepa extracts J. Santas et al. 40
tion with magnetic stirring (Multipoint Magnetic Stir-
rer, SBS) at 900 rpm, the sample was held for 15 min in
an ultrasonic bath (Fungilab Ultrasound), and the
extract was separated from the residue by centrifugation
at 1900 · g. Extraction was repeated increasing the
stirring time to 60 min. Finally, a third extraction was
performed increasing the stirring time to 90 min. The
three supernatants were combined, filtered through a
Whatman No. 2 filter and volume adjusted to 100 mL
with same solvent. Methanol of 50 mL of the resulting
extract was removed at 45 C under vacuum in a
rotatory evaporator and the resulting aqueous extract
was subfractioned with 20 mL of ethyl acetate in order
to obtain hydrophilic and hydrophobic subfractions.
Extraction with ethyl acetate was performed three times.
The aqueous subfraction was made up to an appro-
priate volume with Milli-Q water while the ethyl acetate
subfraction was concentrated to dryness at 45 C under
slight nitrogen flux and finally dissolved in 50% DMSO
in water (v ⁄ v).
Determination of total phenol and flavonoid content
The total phenol content of each extract (TP) was
determined in triplicate by the Folin–Ciocalteu method
as described previously (Santas et al., 2008). Values were
determined from a calibration curve prepared with rutin
1
(ranging from 2 to 25 lmol L) final concentration and
2
R = 0.995). Results are expressed as means ± SD in
lmol of rutin equivalents per g of dry weight (lmol rutin
1
eq. g) D.W.).
Flavonoid content was determined as described by
Bonvehi et al. (2001) with a few modifications. An
appropriate concentration of extract was diluted ten
times with 2% AlCl3 in methanol solution (5% acetic acid
in methanol). The mixture was allowed to react for
10 min. and the absorbance was read at 430 nm against a
sample blank without reactants. Values were determined
from a calibration curve prepared with rutin (ranging
1 2
from 16 to 160 lmol L) final volume and R = 0.998)
1
and expressed as means ± SD in lmol rutin eq. g) D.W.
Antioxidant activity
The method of Re et al. (1999) was used to determine
the Trolox Equivalent Antioxidant Capacity (TEAC) as
a measure of the radical scavenging activity of the
extracts, as described previously (Santas et al., 2008).
The results are expressed as means ± SD in lmol of
1
Trolox g) of D.W.
Antimicrobial activity
Microorganisms
The antimicrobial activity of the extracts and pure
standards was evaluated against the following strains of
2010 The Authors. International Journal2010ofInstitute
Journal compilation Food of Science and Technology
Food Science and
Technology 2010
food spoilage microorganisms obtained from the Cul- 1
140 lg mL) of flavonol standards and 10–100 mg
ture Collection and Research Centre of Valencia (Uni- 1
D.W. mL) of final extract. After 24 h of incubation
versity of Valencia, Spain): B. cereus (CECT 5144), S.
at 30 C, wells were visually examined for evidence of
aureus (CECT 239), M. luteus (CECT 5863), L. mono-
growth. Minimal inhibitory concentration was deter-
cytogenes (CECT 911), E. coli (CECT 99), P. aeruginosa
mined as the lowest concentration where no growth was
(CECT 108) and C. albicans (CECT 1002). Stock observed. Assays were performed in triplicate.
cultures were maintained at 4 C on plates of plate
count agar (PCA, Scharlab, S.L., Spain).
Results and discussion
Disc diffusion assay
Quercetin and kaempferol flavonols were dissolved in Phenol and flavonoid content
1
DMSO (5 mg mL) ). Flavonol standards and extracts The total phenol content (TP) values determined by the
were sterilised by filtration through 0.45 lm Nylon filter Folin–Ciocalteu method are presented in Table 1. The
and its antimicrobial capacity was evaluated by the disc ethyl acetate subfraction of Grano de Oro (GO) variety
diffusion method (Sharififar et al., 2007). showed the highest phenol content (44.37 ± 1.49 lmol
Active cultures used for experiments were incubated 1
of rutin eq. (g D.W.)) ) followed by Fuentes de
in triptone soy broth (TSB, Scharlab, S.L., Barcelona, Ebro (FE) variety (12.59 ± 0.09 lmol of rutin
Spain) overnight at 30 C. Cell density was optically 1
eq. (g D.W.)) ), while Calc¸ ot de Valls (CV) showed
determined at 600 nm in a UV–Vis mini 1240 Shimadzu the lowest TP (1.35 ± 0.10 lmol of rutin eq. (g
spectrophotometer in comparison with previous cali- 1
D.W.)) ). A similar order of activity was observed for the
bration curves with same microorganism. Inocula were aqueous subfractions with values of 23.46 ± 0.84, 18.82
prepared by dilution to achieve cell densities of ± 0.49,
5 1
10 CFU mL) in Ringer (Biokar Diagnostics, Beau- 1
8.36 ± 0.49 lmol of rutin eq. (g D.W.)) for GO >
vais, France). FE > CV, respectively.
Inoculum (500 lL) was mixed with 5 mL of PCA and Flavonoids were mainly present in the ethyl acetate
spread on a fine basal layer of solid media. Flavonol subfractions with values of 34.92 ± 0.75, 17.95 ± 0.16,
1
(10 lL, 5 mg mL) ) or extract (10 lL, 800 mg D.W. 0.38 ± 0.01 lmol of rutin eq. (g D.W.)) for GO >
)1
mL ) were added to 6 mm diameter discs. Gentamicin FE > CV, respectively.
1
sulphate (40 lg mL) ) was used as positive control Results are in good agreement with recent publica-
against bacteria while Amphotericin B (40 lg mL) )
1 tions where flavonoids were mainly present in the ethyl
was used against C. albicans. DMSO was used as acetate subfraction but represented only a small per-
negative control showing no inhibition. centage of the total phenols present in the aqueous
Plates were incubated for 24 h at 30 C and anti- subfraction (Singh et al., 2009). This is consistent with
the low solubility of flavonols in water with quercetin
microbial activity was evaluated by measuring the and kaempferol being the most common flavonoids
inhibition zone against microorganisms. Assays were present in onion extracts (Nuutila et al., 2002; Lanzotti,
performed in triplicate. 2006).
The edible part of yellow onion variety had higher
Micro-well dilution assay total phenol and flavonoid content than white onion
5
Inocula
1
were prepared in order to obtain 10 CFU varieties, while among the white varieties tested FE had
mL) concentrations. Minimal inhibitory concentration
(MIC) of sterilised flavonols and onion extracts was
Table 1 Total phenol, flavonoid content and antioxidant capacity of
determined in ninety-six-well plates with a modified different onion extracts
method based on that previously described (Mann &
Markham, 1998; Delaquis et al., 2002). Inocula (50 lL) Phenol content Flavonoid content TEAC
lmol rutin lmol rutin lmol Trolox g)1
was dispensed into each well, and 20 lL of an appro- )1 )1
priate dilution of the standards or onion extracts was eq. g D.W. eq. g D.W. D.W.
mixed with 130 lL of TSB which contained 0.23% agar Ethyl acetate
(w ⁄ v) and dispensed into the previously inoculated wells. CV 1.35 ± 0.10 0.38 ± 0.01 4.55 ± 0.44
The final concentration of the antimicrobial dilution to FE 12.59 ± 0.09 7.95 ± 0.16 24.59 ± 0.67
be tested was 10% (v ⁄ v) and the final concentration of GO 44.37 ± 1.49 34.92 ± 0.75 74.86 ± 1.77
agar was 0.15% (w ⁄ v). H2 O
CV 8.36 ± 0.49 0.29 ± 0.10 8.74 ± 0.29
Flavonol standards and ethyl acetate subfractions
FE 18.82 ± 0.49 0.78 ± 0.02 26.69 ± 0.00
were diluted with 50% DMSO in water (v ⁄ v). DMSO

final concentration was 5% (v ⁄ v) in each well and the GO 23.46 ± 0.84 2.51 ± 0.11 24.07 ± 0.45

same solvent was added as negative control, showing no Results expressed as means ± SD of triplicate analysis. FE, Fuentes de
inhibition. Concentrations tested ranged from 20 to Ebro; CV, Calc¸ot de Valls, GO, Grano de Oro.
the highest phenolic and flavonoid content as has been
previously reported (Sellappan & Akoh, 2002; Yang
et al., 2004; Santas et al., 2008).
Several publications have reported that flavonoids are
the main group of phenols present in FE white onion
and in yellow onion varieties (Sellappan & Akoh, 2002;
Santas et al., 2008). However, results reveal that Folin–
Ciocalteu method can generate an overestimation of
total phenol content as can be observe in water
subfraction. Other non-phenolic substances such as
vitamin C or sugars, present in the water subfraction,
can interfere with the Folin–Ciocalteu results generating
an overestimation of total phenol content (George et al.,
2005).
Antioxidant capacity
The TEAC assay is one of the most frequent methods
for determination of antioxidant activity. It is based on
the radical scavenging capacity of antioxidants towards
the ABTS radical cation, which is generated by oxida-
tion of ABTS.
The antioxidant capacity of the extracts was consis-
tent with the phenol and flavonoid content being
74.86 ± 1.77, 24.59 ± 0.67, 4.55 ± 0.44 lmol
1
Trol- ox (g D.W.)) for the ethyl acetate
subfraction and
24.07 ± 0.45, 26.691
± 0.00, 8.74 ± 0.29 lmol
Trol- ox (g D.W.)) for the water subfraction for the
GO,
FE and CV varieties, respectively. GO showed the
highest antioxidant capacity due to the presence of a
higher flavonoid content while the antioxidant capacity
and flavonoid content of CV were lower than the values
for the other two varieties. Other publications have
previously reported that coloured onion varieties such as
yellow or red onions have higher antioxidant capacity
than white onion varieties which can be attributed to its
higher flavonoid content (Yang et al., 2004; Stratil et al.,
2006). In consequence, differences in flavonoid content
are well correlated with that of antioxidant capacity.
Antimicrobial activity
The results are presented in Table 2. Quercetin showed a
higher inhibition than kaempferol in the disc diffusion
assay inhibiting all strains of bacteria tested (from
9.8 ± 0.6 to 15.0 ± 1.0 mm), whereas kaempferol was
only effective against the gram positive bacteria S.
aureus and Micrococcus luteus (9.3 ± 1.2 and
10.3 ± 0.6 mm, respectively). In contrast, kaempferol
was more effective than quercetin in inhibiting bacterial
growth of B. cereus, L. monocytogenes and P. aeruginosa
in micro-well dilution assay and resulted as effective as
quercetin in inhibiting the growth of S. aureus and
1
M. luteus with MIC values of 40 lg mL) .
Gram negative bacteria were more resistant than
gram positive bacteria to the effect of the flavonol

Table 2 Antimicrobial activity of flavonols (quercetin and kaempferol) and ethyl acetate and water sub-fractions of onion crude extracts

IZ MIC

FE GO CV
GO CV
Ethyl Ethyl Ethyl Ethyl Ethyl
acetate acetate Quercetin Kaempferol Control H2 O acetate H 2O acetate H2 O acetate Quercetin Kaempferol

Gram ± bacteria
Bacillus cereus 9.5 ± 0.7 8.8 ± 0.4 10.7 ± 0.6 n.d. 14.7 ± 1.7 >80 >100 >80 40 >80 50 80 60
Staphylococcus 10.5 ± 0.5 8.8 ± 0.4 15.0 ± 1.0 10.3 ± 0.6 10.3 ± 0.6 >80 >100 >80 80 >80 80 40 40
aureus
Micrococcus n.d. n.d. 13.3 ± 0.6 9.3 ± 1.2 10.3 ± 1.2 >80 >100 >80 80 >80 100 40 40
luteus
Listeria 12.0 ± 0.0 9.0 ± 0.0 12.2 ± 1.0 n.d. 16.3 ± 0.6 >80 80 >80 40 >80 50 60 40
monocytogenes
Gram - bacteria
Pseudomonas n.d. n.d. 11.8 ± 1.0 n.d. 12.2 ± 0.8 >80 >100 >80 >100 >80 >100 100 80
aeruginosa
Escherichia coli n.d. n.d. 9.8 ± 0.6 n.d. 8.83 ± 1.6 >80 >100 >80 >100 >80 >100 >140 >140
Yeast
Candida albicans n.d. n.d. n.d. n.d. 13.3 ± 0.6 >80 >100 >80 >100 >80 >100 n.d. n.d.

IZ: Disc inhibition zone expressed as means ± SD in mm of 8 mg of D.W. ⁄ disc of extract, 50 lg ⁄ disc of flavonol standard and 0.4 lg mL)1 of antibiotic
standard as positive control.
MIC: minimal inhibitory concentration expressed as means ± SD in mg of D.W. mL)1 of extracts and lg mL)1 of flavonol standards.
FE: Fuentes de Ebro, CV: Calc¸ ot de Valls, GO: Grano de Oro.
n.d.: not inhibition detected. Water subfractions tested showing no inhibition zone are excluded from the table. Bold values denote effective inhibition.
standards. Only quercetin inhibited the growth of P.
aeruginosa (11.8 ± 1.0 mm) and E. coli (9.8 ± 0.6 mm)
in the disc diffusion assay. In micro-well dilution assay
E. coli was resistant to quercetin and kaempferol up to
1
the maximum concentration tested (140 lg mL) ) which
was limited by the solubility of both flavonols in broth
media under the experimental conditions tested. In good
agreement with these results some studies have also
reported the higher resistance of gram negative bacteria
to polyphenols (Taguri et al., 2004), which can be
attributed to the protective effect of the lipopolysaccha-
rides coating layer (Ikigai et al., 1993). Both flavonols
were ineffective in inhibiting C. albicans.
The results agree with those previously reported for
activity against antibiotic-resistant bacteria where quer-
cetin had a higher inhibitory effect than kaempferol
when evaluated in disc assay (Xu & Lee, 2001).
However, assays performed in broth media showed that
kaempferol was more efficient than quercetin in inhib-
iting bacterial growth. The disc assay is a method which
is more dependent on the solubility and diffusion in the
media of the polyphenols tested. Experimentally, ka-
empferol has low solubility in aqueous media which
causes a loss of inhibition in the disc assay in compar-
ison with its effect at appropriate dilution in the broth
media. The disc diffusion assay is commonly used as a
preliminary approach to test antimicrobial potential
(Kloucek et al., 2005). However, these results demon-
strate the importance of performing at least two
different assays since weak inhibition can be observed
in the disc assay for components with low solubility,
although they can be efficient antimicrobial agents when
appropriately diluted in media.
The same tendency was observed in the ethyl acetate
subfraction of the onions extracts. Disc diffusion
method was performed with a concentration of onion
1
extract of 800 mg D.W. mL) which was the maximum
concentration that allowed a good diffusion of the
extract in the media and precipitation was not observed.
Onion extracts were also ineffective in inhibiting C.
albicans. GO and CV inhibited almost all gram positive
bacteria except M. luteus. The growth of more resistant
gram negative bacteria could not be inhibited in either
disc diffusion method or MIC test. Considering the MIC
test (Table 2), GO showed the highest antibacterial
activity of the three varieties tested. The ethyl acetate
subfraction of the crude GO extract was efficient in
inhibiting gram positive bacteria (MIC values ranging
1
from 40 to 80 mg of D.W. mL) ) while the analogous
subfraction of the FE extract showed only antilisterial
1 2008) and the maximum concentration tested was
activity (80 mg of D.W. mL) ) and showed no antimi-
crobial activity against the microorganism tested in the
disc diffusion method. This is in good agreement with
the higher polyphenol and flavonol content of the GO
variety. The ethyl acetate extract from CV showed
antibacterial activity (MIC values ranging from 50 to
1
100 mg of D.W. mL) ) which was slightly less than that
from GO. The antimicrobial activity of CV extracts,
despite the low content of total phenols and flavonoids,
may be due to the presence of different and more active
phenols, or non-phenolic antimicrobial components, e.g.
terpenoids (Cushnie & Lamb, 2005). The aqueous
subfractions of the crude extracts of the three varieties
showed no antimicrobial activity, which can be attrib-
uted to their low flavonoid content. Numerous studies
have reported the presence of flavonols in crude onion
extracts, mainly quercetin and kaempferol which can be
present as glycosides (commonly 3’ and 3’,4’-glycosides)
which are more water soluble. However glycosides are
less active than aglycones, due to the absence of the C-3
hydroxyl group which appears to be critical for antimi-
crobial activity (Xu & Lee, 2001). The ethyl acetate
subfraction is expected to be rich in flavonol aglycones
(Badole & Bodhankar, 2009) which are highly hydro-
phobic and have been previously reported to be good
antimicrobial compounds, which is in good agreement
with the results of this study (Rauha et al., 2000).
On the other hand, the presence in aqueous extracts of
carbohydrates, which may be consumed as a carbon
source by microorganisms, may also contribute to the
low inhibitory effect of the aqueous subfractions.
Gram positive bacteria were relatively sensitive to
onion extracts and flavonol standards while gram
negative bacteria were very resistant (MIC > 100 mg
1
of D.W. mL) ). Listeria monocytogenes was the most
sensitive microorganism being inhibited by FE, CV and
GO ethyl acetate subfractions with MIC values of 80, 50
1
and 40 mg of D.W. mL) . This result is in agreement
with previous literature where L. monocytogenes was
inhibited by the effect of polyphenols such as phenolic
acids (Cushnie & Lamb, 2005), flavonoid aglycones and
glycosides (Vaquero et al., 2007).
No antifungal activity against C. albicans was
observed, which is in agreement with previous reports
where other vegetable extracts rich in flavonoids showed
antimicrobial, but not antifungal activity (Kloucek
et al., 2005). Degirmencioglu & Irkin (2009) also
reported the high resistance of C. albicans against onion
extracts and found that ethanolic extract of white onion
had the most inhibitory effect among the other solvents
tested (water, acetone, diethyleter, methylalcohol). In
our study, methanol 75% in water (v ⁄ v) has been used
as the most efficient solvent for the extraction of onion
flavonoids based on previous publications (Santas et al.,
limited by solubility of the extracts in the media. DMSO
was also used as the most appropriate solvent for
dilution of flavonol and ethyl acetate subfraction of
onion extracts because ethanol and methanol resulted
inhibitory against the microorganism tested even at
lower concentrations. The inhibitory effect of the
extracts and pure standards was very dependent on
experimental conditions. Inocula concentration, nutri-
ent amount and the final dilution of the antimicrobial
used in the test had a great influence on the assay. In
consequence, some extracts reduced the growth of
microorganisms but were not sufficiently effective to
inhibit microbial activity completely, making compari-
son with the literature more difficult since a universal
consensus about methodology is not yet established. In
the present experiment, MIC was confirmed as a good
method to evaluate the antimicrobial activity of com-
pounds with low solubility in water. Addition of an
appropriate concentration of agar reduced flavonoid
precipitation which can result in a reduction of antimi-
crobial activity. Therefore, the method previously
described by Mann & Markham (1998) to determine
the MIC of essential oils is also useful for other
compounds with poor water solubility including flavo-
noids.
In the present study, onion extracts inhibited the
growth of some important bacteria commonly associ-
ated with food deterioration. Odourless lyophilised
onions can be a good source of non-volatile antimicro-
bial compounds and in consequence may be useful to
improve food preservation without undesirable changes
of organoleptic properties.
Conclusion
Onions are known as a good source of flavonoids,
mainly quercetin and its glycosylated derivatives. The
present study confirms the presence of flavonoids in
crude onion extracts and confirms their activity as
antioxidant compounds. Flavonoids were mainly pres-
ent in the ethyl acetate subfraction of the extracts. The
Grano de Oro variety contained the highest amount of
phenols and flavonoids, and had the highest antioxidant
capacity followed by Fuentes de Ebro and Calc¸ ot
de Valls varieties.
The flavonols quercetin and kaempferol were efficient
inhibitors of gram positive bacteria while gram negative
bacteria were more resistant. The yeast C. albicans
showed complete resistance to both flavonols.
The same properties were observed in the ethyl acetate
subfraction of crude onion extracts. Grano de Oro and
Calc¸ ot de Valls extracts were efficient in inhibiting
gram positive bacteria, but not gram negative bacteria
and C. albicans. Fuentes de Ebro was only efficient in
inhibiting L. monocytogenes which were the most
sensitive bacte-
ria. No inhibitory effect of the water subfraction of the
extracts was observed.
In conclusion, these results suggest that the use of
onions as a spice, can be useful not only to increase the
organoleptic quality of food, but also to enhance the sta-
bility and preservation of food systems. Among the
varieties tested, the highest expectations can be focused on
Grano de Oro with the highest flavonoid content.
Acknowledgments
This work was supported by ‘Ministerio de Educacion y
Ciencia’ of Spain as project AGL2005-08088-C02-02.
Authors are grateful to SILLIKER Iberica for the
technical assistance. The authors declare that they have
no conflict of interest.
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