Journal of Medicinal Plants

Evaluation of Antioxidant and Antimicrobial Effects of Chamomile

(Matricaria chamomilla L.) Essential Oil on Cake Shelf Life

Khaki M (M.Sc.), Sahari MA (Ph.D.)*, Barzegar M (Ph.D.)

Food Technology Department, College of Agriculture, Tarbiat Modares

University, Tehran, Iran
*Corresponding author: Food Technology Department, College of Agriculture,
Tarbiat Modares University
Tel: +98 – 21 – 48292328, Fax: +98- 21- 48292200
Email: sahari@modares.ac.ir

Received: 30 Oct. 2011 Accepted: 13 Oct. 2012

Abstract
Background: Essential oils of medicinal plants such as chamomile are very complex natural
mixtures which can contain compounds at quite different concentrations and some of their
components have antioxidant and antimicrobial activities in foodstuff.
Objective: The antioxidant and antimicrobial effects of chamomile essential oil in cake
preparation were evaluated during 75 days of storage.
Methods: Cake were produced: cakes were treated with chamomile essential oil in three levels
(0.05%, 0.1%, 0.15%), cakes were not treated with synthetic and natural antioxidant and
antimicrobial agents, and a commercial cake containing synthetic antioxidant (tert butyl hydro
quinine= TBHQ) and antimicrobial (potassium sorbate) agents. In all samples peroxide value,
thiobarbitoric value, acidity, microbial test (yeast and mould) and sensory test (color, flavor,
texture, taste and overall quality) were determined at time intervals 1, 7, 15, 30, 40, 60 and 75
days.
Results: Statistical results showed that, the sample containing chamomile at 0.15%, had good
antioxidant and antimicrobial activity in comparison with the control samples (without any
synthetic and natural antioxidant and antimicrobial agents) (p<0.01). But its activity was less
than that of synthetic ones (TBHQ and potassium sorbate as antioxidant and antimicrobial agent)
(p<0.01). In sensory evaluation, the sample containing chamomile at 0.05% had higher score in
flavor, taste and overall acceptability than the samples at 0.15 and 0.1% (p<0.05).
Conclusion: Chamomile essential oil as a natural antioxidant and antimicrobial agent can
increase shelf-life of food products and due to absence of synthetic agents are safe with no side
effect on human health.
Keywords: Matricaria chamomilla, Essential oil, Cake, Antioxidant activity, Antimicrobial
activity

9
Volume 11, No. 43, Summer 2012
 Evaluation of …

Introduction flowers and contributes to the observed

Cake manufacturers face a major problem pharmacological properties [6]. Chamazulene
of lipid oxidation and mould growth which extracted from chamomile was shown to
limits shelf-life of their product. The use of inhibit Fe+2/ascorbate-induced lipid
antioxidant and preservatives can reduce this peroxidation [7]. Water and alcohol extracts of
problem [1]. The antioxidants most frequently chamomile flowers showed a moderate effect
used are synthetic phenols such as butylated in controlling hydrolytic rancidity, measured
hydroxy toluene (BHT), butylated hydroxy by peroxide value and free fatty acids [2].
anisole (BHA) and propyl gallate [2]. The composition, structure as well as
However the safety of these synthetic functional groups of the oils play an important
antioxidants and preservatives has been role in determining their antimicrobial activity.
questioned due to toxicity, liver damage and Usually compounds with phenolic groups are
carcinogenicity. Therefore development and most effective [8]. Chamomile oil at 3000 ppm
use of safer antioxidants from natural sources exhibited the highest inhibition against
are of interest because of possible negative Aspergillus flavus, Aspergillus parasiticus and
effects of synthetic food additives on human Fusarium moniliforme [7]. 1
health [3]. Some studies have disclosed the Essential oils of herbals are very complex
potential sources of natural antioxidants for the natural mixtures which can contain about 20-
bakery products [4]. The keeping quality of 60 compounds at quite different concentrations
baked foods such as crackers, cookies and [9]; some of their components have
biscuits is of great economic importance since antioxidant and antimicrobial activities. The
these products are widely used and are often aim of this study was to evaluate antioxidant
stored for extended periods before and antimicrobial activity of chamomile
consumption. The spices most commonly used essential oil (due to the good antioxidant and
in bakery products are cinnamon, mint, mace antimicrobial effects [2, 7]) on cake shelf life.
cloves, poppy and sesame [5]. Several studies
have demonstrated that a variety of plants such
as chamomile, cinnamon possess antioxidant
Material and Methods
and antimicrobial activities. Chamomile Materials
(Chamomile recutita L.) is used for a variety Chamomile essential oil (Matricaria
of herbal remedies and several classes of chamomilla L.) was used as source of natural
biologically activity. Compounds have been antioxidant and antimicrobial preservatives. It
identified from the plant including coumarins was purchased from the Institute of Medicinal
and sesquiterpenes; there have also been many Plants and Natural Products Research in
reports on the identification of flavonoid from Gorgan, Iran (May 2010) and then the
this plant. Apigenin is quantitatively the most essential oil was extracted by steam
abundant flavonoid found in chamomile distillation, using a Clevenger-type apparatus

Journal of Medicinal Plants, Volume 11,

10 No. 43, Summer 2012
13
 Khaki et al.

[10]. The obtained essential oil was dried over were prepared by adding chamomile essential
anhydrous sodium sulphate and kept at 4°C oil at 3 different rates of 0.05% (Ch-0.05),
until it was used. Oil with no antioxidant was 0.1% (Ch-0.1) and 0.15% (Ch-0.15) as natural
purchased from Behshahr factory, flour with antioxidant and antimicrobial agent.
no additive was purchased from Taban
Factory, eggs and baking powder were Methods
purchased from market. Chemicals (of The lipid of ground cakes were extracted
analytical grade) required: hexane, acetic acid using n-hexane by subjected to steam for 30
glacial, chloroform, thiobarbitoric acid, tert min [5]. Stability of cake lipids was
butyl hydro quinine (TBHQ), sodium determined during storage for 75 days [1, 5] at
hydroxide, ethanol, sodium thiosulfate, ambient temperature measuring peroxide
saturated potassium iodide, potassium sorbate, value, thiobarbitoric acid value, free fatty acids
pepton water, DG18 (dicholoran 18% glycerol (FFA) and microbial test (yeast and mould
agar) were obtained from authentic companies. counts). The above analyses were carried out
in three replicates.
Preparation of cake
Cakes were prepared by the following Peroxide value (PV)
method. Sugar (20%) and egg (14%) were Oxidation was periodically assessed by the
mixed for 3-4 minutes, after that flour (26%) measurement of peroxide value (PV) at 1st,
and baking powder (0.5%) were added and 7th, 15th, 30th, 45th, 60th and 75th days of
creamed. The essential oil was blended with storage according to the AOCS method [11].
fat (20%) and mixed with the above cream.
After 5 min of mixing water was added to the Thiobarbitoric acid (TBA)
dough then it was mixed to obtain Thiobarbitoric acid (TBA) values of the
homogenous dough and placed on aluminum samples were measured during the same days
plates, then baked at 20oC for 20 min after of storage according to the AOCS method
baking, plates were allowed to cool, were [12].
covered with air-tight sterile foil and stored in
ambient temperature. Free fatty acid (FFA)
Cake were prepared to provide five Free fatty acid contents were determined
treatments: control treatment 1 received no during 1st, 7th, 15th, 30th, 45th, 60th and 75th
antioxidant or antimicrobial agent (Con 1= days of storage according to the method
negative control samples), control treatment 2 described by Bhanger et al., 2008 [13].
was prepared with addition of TBHQ (100
ppm) as synthetic antioxidant and sodium Yeast and mould counting
sorbate (0.3%) as antimicrobial agent (Con 2= Number of yeast and mould in cakes were
positive control samples), the other variations counting in Dicholoran Glycerol Agar

11
Evaluation of …

(DG18%) medium during 1st, 10th, 30th, 50th, showed that initially peroxide value in all samples
70th days of storage according to the method was low and increased during storage. In Cont 1
of ISO [14]. samples, the rate of hydroperoxides formation
was higher than in the other samples treatments at
Sensory studies every sampling date. Whereas, the increase in PV
Sensory evaluation of cakes was done to in Cont 2 was marginal and was the lowest
determine the acceptability of the cakes among all treatments and samples.
prepared by chamomile essential oil addition In comparison with Cont 1 sample, the
10 days after baking. For this study thirteen cakes containing chamomile essential oil had
panelists were recruited from Food Science lower PV but their peroxide values were
Faculty of Tarbiat Modares University. The higher than those of Cont 2. It was interesting
Seven samples were coded differently and to note that cakes prepared by chamomile
served to panelists. Sensory scores for essential oil at high concentration (0.15%) had
different attributes like color, flavor, texture, the lowest PV during the storage, in fact
taste and overall quality were recorded. The increased of concentration caused decreased
samples were evaluated for sensory attributes PV. The peroxide values after 75 days were
on the tenth day after production as excellent= different significantly among the all samples.
5, good= 4, suitable=3, not suitable = 2 and Changes occurring in TBA and FFA values
ineligible= 1. of the cake variations during storage are given
in Figures 2 and 3. Increase in TBA and FFA
Statistical analysis values were observed in all cake samples by
The experimental data were analyzed time. However cont 1 treatment had the
statistically for variance by using SPSS 18 highest value (11.9) in FFA test and (0.57) in
program. Data recorded as means ± standard TBA test after 75 days.
deviation of three replicate measurements. The TBA values of the samples prepared by
Analyses of variance were performed by chamomile essential oil, ranged from 0.03-0.4
ANOVA test and significance of differences and in the Cont 2 ranged from 0-0.08. The
between the means was determined by values obtained are within acceptable range
Duncan’s multiple range tests. after 75 days but in the Cont 1 sample ranged
from 0.1-0.57.
Results In the cakes prepared by chamomile
Chemical analyses essential oil TBA and FFA values increased by
Peroxide values (PVs) were estimated in all decreasing the chamomile concentration.
cakes to determine the extent of peroxide FFA increased in all samples, it was the
formation due to oxidation of fat lowest in Cont 2 and highest in Cont 1, in
(milliequivalents of O2/kg of fat) during the cakes containing chamomile essential oil was
storage period of 75 days (Figure 1). Data higher than Cont 1 and lower than Cont 2.

Journal of Medicinal Plants, Volume 11,

12 No. 43, Summer 2012
 Khaki et al.

14
a
12

10 a
PV (meq O2/Kg oil)

8 b Ch‐0.05
Ch‐0.1
a
6 Ch‐0.15
c
d Cont1
b
4 a b
Cont2
a bb c c
a c d d
b
2 cd
a bcc
bc d d e e e
e d e
0
1 7 15 30 45 60 75
Days Of Storage

Fig. 1 - PV (meq O2/kg oil) changes of produced cakes samples during 75 days of storage.
The same letters (a, b, c, d and e) are not significantly different (p<0.01), Adding chamomile essential oil at rates of 0.05%
(Ch-0.05), 0.1% (Ch-0.1) and 0.15% (Ch-0.15); Con 1= no antioxidant or antimicrobial agent; Con 2= with addition of TBHQ
(100 ppm) as synthetic antioxidant and sodium sorbate (0.3%) as antimicrobial agent

0.6
a

0.5

b
0.4
a
TBA (m eq MDA/kg oil)

c
Ch‐0.05
a c Ch‐0.1
b
0.3 b Ch‐0.15
a c Cont1
a c Cont2
a c c
0.2
b
a b b

a bb
b b
0.1 b d
d
b c d
b b
c
c c
0
1 7 15 30 45 60 75
Days Of Storage

Fig. 2 - TBA (meq MDA/kg oil) changes of produced cakes during 75 days of storage
The same letters (a, b, c, d and e) are not significantly different (p<0.01), Adding chamomile essential oil at rates of 0.05%
(Ch-0.05), 0.1% (Ch-0.1) and 0.15% (Ch-0.15); Con 1= no antioxidant or antimicrobial agent; Con 2= with addition of TBHQ
(100 ppm) as synthetic antioxidant and sodium sorbate (0.3%) as antimicrobial agent. MDA=malondialdehyde

13
Evaluation of …

14

a
12

10
Free fatty acid (% Of Oleic acid)

8 Ch‐0.05
a Ch‐0.1
Ch‐0.15
6 Cont1
Cont2

4
a

a
2 a
b c b c
b c d d d e
bbb b bcbc b c d e e e
b bcbc a c b c
0
1 7 15 30 45 60 75
Days Of Storage

Fig. 3 - FFA (% oleic acid) changes of produced cakes during 75 days of storage
The same letters (a, b, c, d and e) are not significantly different (p<0.01), Adding chamomile essential oil at rates of 0.05%
(Ch-0.05), 0.1% (Ch-0.1) and 0.15% (Ch-0.15); Con 1= no antioxidant or antimicrobial agent; Con 2= with addition of TBHQ
(100 ppm) as synthetic antioxidant and sodium sorbate (0.3%) as antimicrobial agent

On the whole the changes in TBA and FFA for Aspergillus niger, Monilia sitophila and
the different additive treatments followed exactly species of Mucor and Geotrichum. Among
the same trend by time as did the changes in PVs. these, Penicillium expansum, Penicillium
stoloniferum and Mucor are mycotoxin
Microbial analysis producers [1]. In this study types of moulds
As Figure 4 (mean of ratio of mould and were not identified.
yeast number in samples to Cont 1 sample)
shows in compare to Cont 1 which is with no Sensory analyses
antimicrobial, chamomile essential oil retarded Figure 5 gives the mean scores for sensory
mould growth in cakes. Chamomile essential analysis of cakes on ten days after baking.
oil in high concentration had more effect on There was no significant difference in the
mould, in fact with increasing the texture and color of different cake samples as
concentration of chamomile essential oil evaluated by the panelists. It was observed that
inhibitory activity would increase but its affect cakes prepared by chamomile essential oil in
is less than synthetic ones. After 75 days, some attributes and overall quality received
growth of moulds was observed in all samples. lower score than the other samples. Cakes
Common moulds found in cakes and bakery containing 0.15% chamomile essential oil had
products are Rhizopus stolonifer, Penicillium the lowest score, as shown in Figure 5; you see
expansum, Penicillium stoloniferum,

Journal of Medicinal Plants, Volume 11,

14 No. 43, Summer 2012
 Khaki et al.

1.2

a a a a
1
NO. of mould and yeast in samples/
NO. of mould and yeast in control1

0.8 b

0.6 Ch‐0.05
ab
Ch‐0.1

c Ch‐0.15
0.4
Cont1
b b
b Cont2

0.2 c b c
c
b
a a a a a d d d b d
0
1 10 30 50 70
Days Of Storage

Fig. 4 - Mean of ratios of yeast and mould numbers in samples relative to control (numbers in samples/ numbers
in control 1)
The same letters (a, b, c and d) are not significantly different (p<0.01), Adding chamomile essential oil at rates of 0.05% (Ch-
0.05), 0.1% (Ch-0.1) and 0.15% (Ch-0.15); Con 1= no antioxidant or antimicrobial agent; Con 2= with addition of TBHQ
(100 ppm) as synthetic antioxidant and sodium sorbate (0.3%) as antimicrobial agent

5
a
4.5 a a a a
a a
a a a a
4 a a a
b
M ean scores for sensory analysis 

b b
3.5 b b

c
3
Ch‐0.05
c
2.5 c c Ch‐0.1
d Ch‐0.15
2 d
Cont1
1.5 Cont2

0.5

0
color taste texture flavor Overall quality

Sensory Test 

Fig. 5 - Mean scores for sensory analysis of cakes

The same letters (a, b, c and d) are not significantly different (p<0.05), Adding chamomile essential oil at rates of 0.05% (Ch-
0.05), 0.1% (Ch-0.1) and 0.15% (Ch-0.15); Con 1= no antioxidant or antimicrobial agent; Con 2= with addition of TBHQ
(100 ppm) as synthetic antioxidant and sodium sorbate (0.3%) as antimicrobial agent

15
Evaluation of …

sensory scores increased with decreasing the than would normally be organoleptically
amount of essential oil. The highest scores of acceptable [8]. Our results are in agreement
overall quality belonged to Cont 2. Sensory with those of Al-Ismail and Aburjai, McKay
qualities of Cont 2 (containing commercial and Blumberg, Holly and Patal, Pauli and
preservatives) were more acceptable than all Tolouee et al. [2, 7, 8, 16, 17], because cakes
other treatments. containing 0.15% essential oil had appropriate
antioxidant and antimicrobial activity while
gained lower score for sensory evaluation.
Discussion In order to obtain maximum benefit from
Three different concentrations of use of natural antioxidant and antimicrobial
chamomile essential oil were used in present compounds in food products, several points
study as source of natural antioxidant and should be taken into consideration, for
antimicrobial agent. The effects of instance: αw, temperature, concentration of
antioxidants on the stability of the added fat essential oil and pH should be controlled
were determined by monitoring PV, TBA and during preparation and storage. The combined
FFA periodically under actual storage impact of preservatives commonly used in
conditions (ambient temperature). Cakes bakery products and other important factors
containing synthetic antioxidant and such as water activity (αw), pH on the
antimicrobial (TBHQ and potassium sorbate) prevention of fungal spoilage has been
chemicals possess stronger activity than the assayed. Potassium sorbate is a suitable
natural ones but in comparison to samples with preserving agent that inhibits growth of
no synthetic antioxidant or antimicrobial xerophilic fungi in bakery products of pH near
agents, chamomile essential oil had significant 4.5 regardless of αw level. For products
activity (p<0.01) and the effect of chamomile slightly acidic the addition of this preservative
essential oil increased with increasing must be combined with low water activity
concentration. levels otherwise other additional controlling
Our results revealed that chamomile factors must be applied [18].
essential oil applied to cake was fungicidal and In fact hurdle technology is useful to
retarded mould growth. control oxidation and spoilage due to
Antioxidant activity of plants is mainly due to microorganisms in bakery products of
the presence of phenolic compounds [2]. intermediate moisture.
Chamomile possesses different phenolics. Some Plant-derived essential oils due to their
phenolic compounds reportedly have both content of antimicrobial compounds possess
antioxidant and antimicrobial effects [1, 15]. adequate potentials as natural agents for food
When essential oils are used in food preservation. Their antimicrobial activity is
production systems amounts required are high assigned to a number of small terpenoid and
and as such they are often higher in quality phenolic compounds [19].

Journal of Medicinal Plants, Volume 11,

16 No. 43, Summer 2012
 Khaki et al.

Conclusion water activity, temperature, pH and packaging

In conclusion, the above results showed conditions to reduce the amount of food
that, the sample containing chamomile at preservatives. Natural antioxidants and
0.15% had better antioxidant and antimicrobial antimicrobials can increase shelf-life of food
activities than the samples without any products and due to absence of synthetic
synthetic and natural antioxidant and agents are safe with no side effects on human
antimicrobial agents. But its activity was less health.
than that of TBHQ and potassium sorbate as
antioxidant and antimicrobial agent (synthetic Acknowledgment
ones). Results show that essential oils alone The authors thank the financial support of
cannot provide complete protection against Tarbiat Modares University Research Council
oxidation and microbial contamination in and Center of Excellency for Recycling and
cakes. It is therefore necessary to control the Losses of Strategic Agricultural Products.

References
1. Lean LP, Mohamed S. Antioxidative and and their application in biscuits. Journal of
antimycotic effects of turmeric, lemon-grass, Food Chem. 2005; 90: 317 - 21.
betel leaves, clove, black pepper leaves and 6. Švehlíková V, Bennett NR, Mellon AF,
Garcinia atriviridis on butter cake. Journal of Needs WP, Piacente S, Kroon AP, Bao Y.
Agriculture and Food Science 1999; 79: 1817 Isolation, identification and stability of
- 22. acylated derivatives of apigenin 7-O-glucoside
2. Al-Ismail KM, Aburjai T. Antioxidant from chamomile (Chamomilla recutita L.
activity of water and alcohol extracts of Rauschert). Journal of Phytochem. 2004; 65:
chamomil flowers, anise seeds and dill seeds. 2323 - 2.
Journal of Agriculture and Food Sci. 2004; 84: 7. McKay DL, Blumberg JB. A review of the
173 - 8. bioactivity and potential health benefits of
3. Nanditha BR, Jena BS, Prabhasankar P. chamomile tea (Matricaria recutita L.).
Influence of natural antioxidants and their Journal of Wiley Interscience 2006; 20: 519 -
carry-through property in biscuit processing. 30.
Journal of Agriculture and Food Sci. 2009; 89: 8. Holley RA, Patel D. Improvement in shelf-
288 - 98. life and safety of perishabale foods by plant
4. Lu TM, Lee CC, Mau JL, Lin SD. Quality essential oils and smoke antimicrobials.
and antioxidant property of green tea sponge Journal of Food Microbiol. 2005; 22: 273 - 92.
cake. Journal of Food Chem. 2010; 119 (3): 9. Bakkali F, Averbeck S, Averbeck D,
1090 - 95. Idaomar M. Biological effects of essential oils-
5. Reddy V, Urooj A, Kumar A. Evaluation A review. Journal of Food and Chemical
of antioxidant activity of some plant extracts Toxicol. 2008; 46: 446 - 75.

17
Evaluation of …

10. Ayoughi F, Barzegar M, Sahari MA, selected pathogenic and beneficial gut
Naghdibadi M. Chemical compositions of bacteria. Journal of Applied Microbiol. 2006;
essential oils of Artemisia dracunculus L. and 100: 296 - 305.
Matricaria chamomile and evaluation of their 16. Pauli A. α-Bisabolol from chamomile-A
antioxidative effect. Journal of Agricultural specific ergostrol biosynthesis inhibitor.
Science and Technol. 2011; 13: 79 - 88. Journal of Aromatherapy 2006; 16: 21 - 5.
11. AOCS. In: D. Firestone, (Ed), Official 17. Tolouee M, Alinezhad S, Saberi R,
Methods and Recommend Practices of Eslamifar A, Zad SJ, Jaimand K, Taeb J,
American Oil Chemist Society. 4thEd. Rezaee MB, Kawachi M, Shams-Ghahfarokhi
Champaign: Cd 8b - 90, 1989. M, Razzaghi-Abyaneh M. Effect of Matricaria
12. AOCS. 2-Thiobarbitoric acid value direct chamomilla L. flower essential oil and
method. Cd 19 - 90, 2006. ultrastructure of Aspergillus niger Van
13. Bhanger MI, Iqbal S, Anwar F, Imran M, Tieghem. Journal of Food Microbiol. 2010;
Akhtar M, Zia-ul-Haq M. Antioxidant 139: 127 - 33.
potential of rice bran extracts and its effects on 18. Guynot ME, Ramos AJ, Sanchis V, Marin
stabilization of cookies under ambient storage. S. Study of benzoate, propionate and sorbate
International Journal of Food Science and salts as mould spoilage inhibitors on
Technol. 2008; 43, 779 - 86. intermediate moisture bakery products of low
14. ISO 2527-1: Microbiology of food and pH (4.5-5.5). Journal of Food Microbiol.
animal feeding Stuffs-Horizontal method for 2005; 101: 161 - 8.
the enumeration of yeasts and moulds- part 1: 19. Kotzekidou P, Giannakidis P, Boulamatsis
Colony count technique in products with water A. Antimicrobial Activity of some plant
activity greater than 0.95, 2008. extracts and essential oils against food borne
15. Si W, Gong J, Tsao R, Zhou T, Yu H, pathogens in vitro and on the fate of inoculated
Poppe C, Johnson R, Du Z. Antimicrobial pathogens in chocolate. Journal of
activity of essential oils and structurally Lebensmittel Wissenschaft Technologie 2008;
57
related synthetic food additives towards 41: 119 - 27.

Journal of Medicinal Plants, Volume 11,

18 No. 43, Summer 2012