American-Eurasian J. Agric. & Environ. Sci.

, 14 (10): 1116-1124, 2014

ISSN 1818-6769
IDOSI Publications, 2014
DOI: 10.5829/idosi.aejaes.2014.14.10.8648

Phenolic Content, Antioxidant and Antimicrobial

Activities of Egyptian and Chinese Propolis
1
S.A. El Sohaimy and 2S.H.D. Masry

1
Food Technology Department, Arid Land Cultivation Research Institute,
City of Scientific Research and Technological Applications, Universities
and Research Centers District, New Borg El Arab, 21934 Alexandria, Egypt
2
Plant Protection and Molecular Diagnosis Department, Arid Land Cultivation Research Institute,
City of Scientific Research and Technological Applications, Universities
and Research Centers District, New Borg El Arab, 21934 Alexandria, Egypt

Abstract: Propolis is a resinous mixture that honeybees collect from tree buds, sap flows, or other botanical
sources. It is used as a sealant for unwanted open spaces in the hive. Propolis is sticky at and above room
temperature, 20C (68F). At lower temperatures, it becomes hard and very brittle. The aim of this study is to
explore the phenolic contents and identify of the Egyptian and Chinese propolis and their biological activity
potentiality, especially antioxidant and antimicrobial activity. Egyptian and Chinese propolis contained
considerable amounts of phenolic compounds. The Egyptian propolis contains phenolic content a little bit
greater than Chinese propolis. The Egyptian propolis showed an antioxidant activity higher than Chinese.
IC50 of Egyptian propolis was (73.49 g/ml) and (81.67 g/ml) for Chinese propolis, whereas the IC 50 for
L-Ascorbic acid as positive control was (39.62 g/ml). The HPLC analysis of Egyptian and Chinese propolis
approved reasonable and different concentrations of phenolic compounds in both Egyptian and Chinese
Propolis. The Egyptian propolis contains high concentration levels of tannic acid (10.64 g/g), catechol
(8.12 g/g) and caffeic acid (7.435 g/g). The Egyptian propolis showed a highest toxicity against Bacillus
subtilis DB 100 host and Streptococcus sp. (IZD= 18 and 20 mm) respectively. On the other hand, Chinese
propolis showed a highest antimicrobial activity and toxicity against Candida albicans and Bacillus subtilis
(IZD=20mm) for both strains.

Key words: Honey Bee Propolis Antioxidant Phenolic Content Antimicrobial

INTRODUCTION this contributes to maintaining the hive inner temperature

at around 35C. Also, bees use it to seal cracks in hives,
Propolis (bee glue) is the generic name for the encapsulate invader carcasses, repairing combs and
resinous product of complex composition collected by strengthening the thin borders [1, 2, 5]. This is obviously
Apis mellifera bees from bud and exudates of various important to protect the hive from a widespread
plants mixed with bee secretions and beeswax. Physically microorganisms specially bacteria and fungal infection.
its a sticky material in room temperature but becomes Propolis was very well known in ancient Egypt, to the
hard and brittle at low temperature [1-3]. The color may be priests who had monopolized medicine, chemistry and art
cream, yellow, green, light or dark brown. Some samples on mummifying corpses [6]. The fact that propolis was
have a friable, hard texture, while other samples may be also known to the old Greeks is demonstrated by the
elastic and gummy [4]. Honeybees utilize propolis to very Greek name of it Makashvili [7]. Abu Ali bin
diverse purposes, among them to seal openings in the Sina (Avicenna) distinguishes two kinds of wax in his
hive. In addition to avoiding the entrance of intruders, well-known work, the clean and the black wax [6].

Corresponding Author: El Sohaimy S. A, Food Technology Department, Arid Land Cultivation Research Institute,
City of Scientific Research and Technological Applications, Universities and Research Centers District,
New Borg El Arab, 21934 Alexandria, Egypt. E-mail: [email protected].
1116
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

The clean wax is that which composes the comb wells MATERIALS AND METHODS
where the bees rear the brood and store the honey and
the black is the filth the hive. It is clear enough that the Samples Collection: Egyptian propolis samples collected
black wax is propolis that after Avicennas testimony from middle delta region, Egypt and the Chinese propolis
[8]. The chemical variability of propolis is, of course, due samples collected from Anhui, China.
to its plant origin, collecting geographic locations the
source plants might vary with respect to the local flora at Sample Preparation: The propolis sample (20 g) was
the site of collection and seasons [4, 9- 11]. It is now extracted with 90% ethanol (200 mL) by mixing for 24 h at
generally accepted that bees collect resinous plant room temperature in dark place. The crude extract was
materials, produced by a variety of botanical processes, recovered by centrifugation (3000 g, 10 min) and dried
in different parts of plants. These are substances actively under vacuum using a rotary evaporator.
secreted by plants, as well as substances exuded from
wounds in plants; they include lipophilic materials on Total Phenolic Content (TPC): The total phenolic
leaves and leaf buds, mucilage, gums, resins and latices compounds assay was carried out using the Folin-
[12, 13]. The specificity of local flora is responsible for the Ciocalteu reagent, following the method of [30] and based
chemical composition of propolis [14]. Propolis is typically on the reduction of a phosphowolframate-
composed of resin and vegetable balsams (50-70%), phosphomolebdate complex by phenolics to blue reaction
essential and aromatic oils and beeswax (30-50%), pollen products. 1mg propolis extract was dissolved in 1ml
(5-10%) and other constituents which are amino acids, methanol and 500 l of dissolved sample was taken
minerals, vitamins A, B complex, E and the highly active and added to 0.5 ml of distilled water and 0.125 ml of
bio-chemical substance known as bioflavenoid Folin-Ciocalteu reagent. The mixture was shaken and
allowed to stand for 6 minutes before addition of 1.25 ml
(Vitamin P), phenols and aromatic compounds [15-17].
of 7% Na2CO3. The solution was adjusted with distilled
The chemical compositions and biological activities of
water to a final volume of 3 ml and mixed thoroughly.
propolis are attributed to plant sources, geographical area
After incubation in the dark for 30 min, the absorbance at
and collecting season [9, 18]. More than 300 components
650 nm was read versus the prepared blank. A standard
have been identified in propolis samples. Flavonoids,
curve was plotted using different concentrations of Gallic
aromatic acids, diterpenoid acids, triterpenoids and
acid (standard, 0-1000 g/mL). Total phenolic content was
phenolic compounds are the major components of
estimated as g Gallic acid equivalents (GAE)/g of dry
propolis [19-23]. In Mediterranean, propolis from Algeria,
weight sample.
Croatia, Cyprus and Greece has a poplar-type chemical
profile, while samples from Crete and South Greece are
Determination of Antioxidant Activities
rich in diterpenes [24]. However, [25] mentioned that the DPPH Radical-Scavenging Activity: DPPH radical -
major compounds of Ethiopian propolis were scavenging activity was measured by direct hydrogen
triterpenoids. Aliphatic acids, aromatic acids, alcohols, donation to the DPPH radical, as previously reported, with
phenols, esters and other compounds were found in minor modifications [30]. For each sample, different
the Egyptian propolis and commercial one [26]. concentrations ranging from 5 to 200 g/mL were prepared
They identified fifty-seven compounds in Egyptian with methanol. The reaction mixtures in the 96-well
propolis, while a total of forty-four compounds have been plates consisted of sample (100 l) and DPPH radical
tentatively identified in commercial propolis. Propolis has (100 l, 0.2 mM) dissolved in methanol. The mixture was
a wide range of biological activity and pharmacological stirred and left to stand for 15 min in dark. Then the
effects as antibacterial and antifungal activity; therefore absorbance was measured at 517 nm against a blank. All
it is the defense of bees against infections [21]. It has determinations were performed in triplicates. The
potential to uncover new biologically active compounds percentage scavenging effect was calculated as:
with important pharmacological effects, especially
antibacterial, antiviral, anti-inflammatory, antitumor, % Inhibition = [1 - (A1 - A2) / A0] 100%
antioxidant, anticancer substances and new bioactive
molecules [2, 10, 27-29]. The aim of this study is to explore where: A0 is the absorbance of the control (without
and identify the phenolic contents of the Egyptian and sample) and A1 is the absorbance in the presence of the
Chinese propolis and their biological activity potentiality, sample, A2 is the absorbance of sample without DPPH
especially antioxidant and antimicrobial activity. radical. The scavenging ability of the samples was

1117
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

expressed as IC50 value, which is the effective Nutrient broth was used to obtain the viable growth of
concentration at which 50% of DPPH radicals were microbes from their freeze-dried form. After 48 h,
scavenged. The IC50 values were calculated from the turbidity in test tube confirmed the growth of microbes
relationship curve of scavenging activities (%) versus that was compared and adjusted to McFarland 0.5
concentrations of respective sample [31, 32]. turbidity standard (108 colony-forming units per milliliter)
[35, 36].
HPLC Analysis of Phenolic Compounds: The phenolic
compounds of the propolis samples were analyzed using Preparation of Propolis Extract for Antimicrobial Test:
high-performance liquid chromatography (HPLC) Ten grams of propolis powder was added to 100 ml of
according [33]. Fifty (50) milligrams of propolis were DMSO (an inert solvent) and kept at a cool and dark place
extracted using 200 ml of ethanol at room temperature for in an amber colored bottle [37]. Agar well diffusion assay
30 minutes. The extract was filtered through a paper filter was carried out to evaluate the antimicrobial potential of
and using methanol, the volume was adjusted to 10 ml. propolis [38]. Petri dishes containing 100 ml of brain heart
One milliliter of this sample was mixed with 0.5 ml Milli Q infusion broth supplemented with 5 ml of 5% sheep blood
water and centrifuged for 3 minutes at 13000 rpm and the were inoculated with approximately 100 l of the
supernatant was used directly for HPLC analysis. respective microbial strain using swab technique.
Each propolis sample was extracted and analyzed in Wells of 8 mm diameter were cut into solidified agar media
triplicate. Phenolic compounds were analyzed using HPLC using a sterilized device. One hundred microliters of the
(Agilent, Series 1100, Germany), an instrument containing propolis extract was poured in the wells and the plates
a binary pump (G1316A), The column used was Zorbax, were incubated at 37C for 48 h. To ensure the
SB-C18, 4.6 x 75 mm with 3.5 m particle size. The elution consistency of all findings, the experiment was
solvents were aq. 1.5% tetrahydrofuran + 0.25% performed and repeated under strict aseptic conditions.
orthophosphoric acid (A) and 100% methanol (B). The antibacterial activity of propolis extract was
The samples were eluted according to the following expressed in terms of the mean of diameter of inhibitory
gradient: 0-5 min 100% A; 5-10 min 85% A, 15% B; 10-20 zone (in millimeters) produced by the extract at the end of
min 70% A, 30% B; 20-40 min 50% A, 50% B; 40-75 min incubation period [30].
50% A, 50% B; 75-80 min100% B. The flow rate was 2
ml/min and the autoinjection volume was 20 l. Determination of Minimum Inhibitory Concentration:
The temperature of the column and injector was Minimum inhibitory concentration (MIC) is defined, as the
+30C and +20C, respectively. The HPLC runs were lowest concentration of extract at which there will be no
monitored at 220 and 320 nm. Analyzed secondary visible growth of the test organism. In the present study,
metabolites were quantified against commercial MIC was determined using serial tube dilution
standards. The identification of the compounds was technique. The MIC of propolis for Egyptian and
based on the HPLC-MS-identification or on comparison Chinese propolis was conventionally determined in
of retention times and spectral characteristics as triplicate for each strain by the macrodilution broth
described in [34]. The quantification of the phenolic method as described by the National Committee for
compounds is based on the commercial standards: Clinical Laboratory Standards (NCCLS) [39, 40]. Serial two
chlorogenic acid; ferulic acid cinnamic acid, p -OH- fold dilutions of propolis extract were prepared in
cinnamic acid, caffeic acid, benzoic acid, vanillic acid, macrodilution tubes and inoculated with constant
apigenin, Pinocembrin, Chlorogenic acid, Acacetin, Gallic amount of test bacteria and then all the test tubes
acid, Itaconic acid, Protocatechoic acid, Catechin, were incubated at 37C for 1824 h. Each tube was mixed
Esculetin, Catechol, Tannic acid, Ferulic acid and and examined for growth, comparing each tube to the
Pyrogallol. control. For each test, DMSO was used as the control
solvent.
Determination of Antimicrobial Activity
Bacterial Strains: The tested bacterial strains in this Statistical Analysis: Triplicate determinations, mean and
study were Candida Albicans, Bacillus Subtilis DB 100 standard deviation were calculated. Calibration curve of
host, Salmonella senftenberg and Streptococcus sp. standard was obtained for concentration vs. absorbance.
(Microbiological Resource Center (MIRCEN), Faculty All data were subjected for analysis using independent
of Agriculture, Ain-Shams University, Cairo, Egypt). variable t-test.

1118
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

RESULTS AND DISCUSSION Table 1: Total phenolic content of Egyptian and Chinese propolis
(expressed as mean of triplicates SD) (P>0.05)
Propolls extract TPC Conc. gGAE/g sample
Total Phenolic Content: Total phenolic content in the
EG 137.520.003
propolis extract was carried out using the Folin-Ciocalteu
CH 123.080.005
reagent and the obtained results confirmed that, the
Egyptian and Chinese propolis contains considerable
Table 2: Antioxidant activity of Egyptian and Chinese Propolis (The
amounts of phenolic compounds. Total phenolic content values mentioned are the means of triplicates SD) (P>0.05)
in propolis extracts were 137.520.003 and 123.080.005 g % Inhibition
GAE/g propolis extract for Egyptian and Chinese propolis Sample Conc. ---------------------------------------------------------------------
respectively (Table 1). The Egyptian propolis contains (g/ml) Egyptian Chinese Ascorbic acid
phenolic compounds a little bit more than Chinese 5 9.340.03 8.980.09 36.280.16
propolis. Actually there is no significant difference 10 12.870.04 12.170.16 49.680.15
between the Egyptian and Chinese propolis in the total 20 25.760.02 25.530.02 57.410.08

phenolic content. The obtained results agree with some 40 36.710.12 34.630.31 64.760.13
60 46.230.23 38.240.22 69.430.25
previously published works, which studied the phenolic
80 55.610.36 49.630.18 75.210.32
content of propolis [29, 41, 42]. The total amount of the 100 63.490.28 56.380.07 86.340.05
phenolic compounds in Finnish propolis ranged from 79.8 120 80.150.31 78.310.19 91.340.36
to 156.3 g/g, the average being 119.5 g/g [41]. There are 140 89.70.06 89.450.37 98.870.29
many limiting factors affecting on the concentration of 160 95.560.34 93.760.51 99.320.41
phenolic compounds, type of solvents, extract 180 98.360.04 97.860.42 99.890.32
temperature, stirring and the origin and source of the 200 99.200.28 99.130.06 99.960.36
propolis [41-43]. This considerable content of phenolic IC50 73.490.39 81.670.28 39.620.34

compounds in either Egyptian or Chinese propolis makes

Table 3: HPLC analysis of phenolic compounds of propolis (P<0.05)
it very important to human health and confirms a wide
Conc. g/g
spectrum of its health benefits. ---------------------------------------------------
Phenolic Compound Egyptian Chinese
Antioxidant Activity: From the received data (Table 2), Cinnamic acid 0.0920.32 0.1670.25
both Egyptian and Chinese propolis showed a high Vanillic acid 3.050.53 0.630.51
Chlorogenic acid 0.280.51 0.0340.56
antioxidant activity potentiality. The Egyptian propolis
p-OH-cinnamic acid 0.0890.02 0.1250.63
showed an antioxidant activity a little bit higher than Benzoic acid 2.310.34 0.910.24
Chinese but there is no significant differences between Pinocembrin 1.1200.26 0.6320.25
them. IC50 of Egyptian propolis was 73.49 g/ml and 81.67 Caffeic acid 7.4350.36 1.2870.63
g/ml for Chinese propolis, whereas the IC50 for L- Apigenin 0.320.34 1.540.18
Chlorogenic acid 4.220.28 0.810.29
Ascorbic acid as positive control was 39.62 g/ml. DPPH-
Acacetin Nil 0.210.76
stable free radical scavenging activity (% inhibition) of Gallic acid 6.350.18 4.360.71
both Egyptian and Chinese propolis extracts and L- Itaconic acid 2.510.42 6.120.48
ascorbic acid increased as the concentration of propolis Protocatechoic acid 0.190.71 2.340.61
extract and L-ascorbic acid were increased (Table 2). L- Catechin 4.220.19 3.660.51
Esculetin 6.390.36 4.690.38
ascorbic acid more effective than propolis at lower Catechol 8.120.25 2.500.21
concentrations (5-60 ug/ml), nevertheless, the propolis Tannic acid 10.640.81 4.130.24
showed antioxidant activity near L-ascorbic acid at higher Ferulic acid 5.140.54 0.190.53
concentrations (80-200ug/ml). These obtained results Pyrogallol 1.960.26 6.320.23
agreed with some published work about Egyptian and
Chinese propolis. Ethyl acetate fraction of Chinese Egyptian provinces (Fayoum; Assiut; Souhag; Dakahlia;
propolis showed significant antioxidant and free Sharkia and Ismailia) possess a good anti-oxidative
radical-scavenging capacities, phenolics contributed to potential [45-47]. An antioxidant is a molecule that inhibits
the antioxidant activity of propolis collected in Anhui, the oxidation of other molecules. Oxidation is a chemical
China. Therefore, Chinese propolis and its phenolics reaction that transfers electrons or hydrogen from a
might be used as a natural antioxidant [44]. All substance to an oxidizing agent. Oxidation reactions can
investigated propolis samples collect form different produce free radicals. In turn, these radicals can start

1119
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

Table 4: Antibacterial activities of Propolis, ampicillin and DMSO against acid) in the Egyptian propolis higher than that in Chinese
various indicator bacteria. (+)= Inhibition zone detected, (-) = No
inhibition zone detected. The values mentioned are the means of
one. In contrary, the concentration of (Cinnamic acid,
triplicates SD p-OH-cinnamic acid, Apigenin, Acacetin, Itaconic acid
and Pyrogallol) in Chinese propolis higher than that in
Egyptian one. These differences in the concentration of
phenolic compounds may cause the differences in
antioxidant activities between Egyptian and Chinese
propolis. The Egyptian propolis contained high
concentrations of Tannic acid (10.64 g/g), Catechol
(8.12 g/g) and Caffeic acid (7.435 g/g). The Egyptian
propolis was analyzed by GC-MS and 25 compounds were
identified, seven compounds were identified in Egyptian
propolis for the first time [45]. The constituents were
phenolic acid esters (72.7 %); phenolic acids (1.1 %);
aliphatic acids (2.4 %); dihydrochalcones (6.5 %);
Chalcones (1.7 %); flavanones (1.9 %); flavones (4.6 %)
and tetrahydrofuran derivatives (0.7 %) [44, 45].

Antimicrobial Activity: The antimicrobial activity was

measured in terms of diameter of the inhibitory zones in a
soft agar layer. From the obtained results in Table (4),
both Egyptian and Chinese propolis showed a reasonable
antimicrobial activity against tested strains (Candida
albicans, Bacillus subtilis DB 100 host, Salmonella
Senftenberg, Streptococcus sp.). The Egyptian propolis
showed a highest toxicity against Bacillus subtilis DB
chain reactions. When the chain reaction occurs in a cell, 100 host and Streptococcus sp. (IZD= 18 and 20 mm)
it can cause damage or death to the cell. Antioxidants respectively. On the other hand Chinese propolis extract
terminate these chain reactions by removing free showed a highest antimicrobial activity and toxicity
radical intermediates and inhibit other oxidation reactions. against Candida albicans and Bacillus subtilis
They do this by being oxidized themselves, so (IZD=20mm) for both strains. In contrary, both Egyptian
antioxidants are often reducing agents such as thiols, and Chinese propolis showed the lowest antimicrobial
ascorbic acid, or polyphenols. The antioxidant activity of activity against Salmonella senftenberg (IZD=10 and 8
propolis may due to the ability of phenolic compounds to mm) respectively. The moderate toxicity recorded against
donate hydrogen ions that can attack the free radicals to Candida albicans for Egyptian peoples (IZD= 16mm) and
prevent the oxidation reactions in the cell and preventing Streptococcus sp. for Chinese propolis (IZD= 12mm).
the oxidation and deterioration of food substances during Furthermore, both Egyptian and Chinese propolis showed
storage as well. The high antioxidant activity of propolis toxicity against all tested microbial strains higher than
makes it a good natural antioxidant that can use as a ampicillin as standard antibiotic (Table 6), the minimum
natural preservative and/or food additives to help guard inhibitor, concentrations (MIC) was calculated for both
against food deterioration. Egyptian and Chinese propolis and the results recorded
in (Table 5). Several researchers have studied the
HPLC Analysis of Phenolic Compounds: The HPLC antimicrobial activity of propolis. The antibacterial activity
analysis of Egyptian and Chinese propolis approved of six propolis solutions from different geographic
reasonable and different concentrations of phenolic locations was active against various bacterial strains
compounds in both Egyptian and Chinese Propolis. [48]. Another in vitro investigation also demonstrated
Concentrations of some phenolic compounds (Vanillic the antimicrobial activity of Brazilian propolis against
acid, Chlorogenic acid, Benzoic acid, Ferulic acid, various periodontopathogens including Pg and Aa [49].
Pinocembrin, Caffeic acid, Chlorogenic acid, Gallic acid, The Inhibitory activity of Brazilian propolis was
Catechin, Esculetin, Catechol, Tannic acid and Ferulic investigated on Aa, Fusobacterium nucleatum, Pg and

1120
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

Table 5: Minimum inhibitory Concentration (MIC) of propolis extract

Antibacterial activity against indicator strain expressed in MIC (mg/ml)
-------------------------------------------------------------------------------------------------------------------------------------------------------
Substances Candida albicans Bacillus subtilis Salmonella senftenberg Streptococcus sp.
Egyptian propolis 9.31.03 9.30.97 37.51.03 3001.02
Chinese propolis 9.31.21 9.31.05 3001.02 3001.03
Ampicillin 3000.86 3001.03 9.31.04 1501.03
DMSO - - - -

Table 6: Antimicrobial activity of propolis extract. (-)= No inhibition zone detected. The values mentioned are the means of triplicates SD
Inhibition zone diameter (mm)
--------------------------------------------------------------------------------------------------------------------------------------
Strain Egyptian Chinese Ampicillin DEMSO
Candida albicans 160.16 200.43 80.39 -
Bacillus subtilis D 100 host 180.13 200.35 100.34 -
Salmonella senftenberg 100.16 80.27 80.18 -
Streptococcus sp. 200.51 120.13 100.37 -

Prevotella intermedia and found that all of the assayed CONCLUSION

bacterial species were susceptible to propolis extract [50].
The antimicrobial activity of Chinese propolis was Propolis is a resinous mixture that honeybees
investigated and approved that the Chinese propolis collect from tree buds, sap flows, or other botanical
exhibited the maximum inhibitory zone of 25 mm for sources. The composition of propolis varies from hive to
Pg and 14 mm for Aa at a concentration of 0.1 g/ml and hive, from district to district and from season to season.
observed that propolis extract had a wide spectrum Propolis has a wide range of biological activity and
antimicrobial activity against Candida albicans and pharmacological effects as antibacterial and antifungal
Bacillus subtilis [51]. All Egyptian propolis samples activity; therefore it is the defense of bees against
showed an inhibition in the growth of all examined infections. It has potential to uncover new biologically
bacteria but the inhibition varied according to the propolis active compounds with important pharmacological effects,
origin [52, 42]. Evaluation of Egyptian propolis as especially antibacterial, antiviral, anti-inflammatory,
immunostimulant, antiviral, antibacterial and antifungal antitumor, antioxidant, anticancer substances and new
agents were done and showed that the Egyptian propolis bioactive molecules. The aim of this study is to explore
has such activities [46]. The differences in the level of the the phenolic compounds in the Egyptian and Chinese
effectiveness of propolis extract as antimicrobial agent propolis and their antimicrobial potentiality. Egyptian and
may refer to the differences of the concentration, types Chinese propolis contained a considerable amount of
and carrier of phenolic compounds. The anti-bacterial phenolic compounds. The Egyptian propolis contains
activity of propolis extract might be due the ability of phenolic content a little bit greater than Chinese propolis.
phenolic compounds to bind to bacterial cell walls and Actually there is no significant difference between the
prevent cell division and growth [53, 30]. The mechanism Egyptian and Chinese propolis in the total phenolic
of antifungal activity of phenolic compounds may refer to content. The Egyptian propolis showed an antioxidant
disruption of Ca2+ and H+ homeostasis, up- and down- activity a little bit higher than Chinese but there is no
regulation of gene transcription similar to Ca2+-stress and significant differences between them. IC 50 of Egyptian
nutrient starvation [54], disruption of membrane integrity propolis was 73.49 g/ml and 81.67 g/ml for Chinese
and impairment of ergosterol biosynthesis in Candida propolis, whereas the IC50 for L-Ascorbic acid as
strains [55]. It is thus clear that both types of propolis positive control was 39.62 g/ml. The HPLC analysis of
Egyptian and Chinese have a good effectiveness as an Egyptian and Chinese propolis approved reasonable and
anti-microbial, which nominated as a natural material that different concentrations of phenolic compounds in both
can be used in food preservation. More extended research Egyptian and Chinese Propolis. The Egyptian propolis
work needed to prepare an antimicrobial product from contained high concentrations of Tannic acid (10.64
propolis. g/g), Catechol (8.12 g/g) and Caffeic acid (7.435 g/g).

1121
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

The Egyptian propolis showed a highest toxicity against 11. Guo, S., S. Fu, Z. Shen, Z. Zhang and Q. Xu, 2011.
Bacillus subtilis DB 100 host and Streptococcus sp. Chemical composition, biological activity and
(IZD= 18 and 20 mm) respectively. On the other hand, application in animal science of propolis- A review.
Chinese propolis extract showed a highest antimicrobial International Conference on Agricultural and
activity and toxicity against Candida albicans and Biosystems Engineering Advances in Biomedical
Bacillus subtilis (IZD=20mm) for both strains. Finally we Engineering, 1-2: 98-101.
can conclude that the Egyptian and Chinese propolis 12. Crane, E., 1988. Beekeeping: Science, Practice and
contains considerable concentrations of phenolic World Resources. Heinemann press, London,
compounds that lead to their biological activity as pp: 367-372.
antimicrobial agent. 13. Silva, B.B., P.L. Rosalen, J.A. Cury, M. Ikegaki,
V.C. Souza, A. Esteves and S.M. Alencar, 2008.
REFERENCES Chemical composition and botanical origin of red
propolis, a new type of Brazilian Propolis. Evidence-
1. Ghisalberti, E.L., 1979. Propolis: a review. Bee World, Based Complementary and Alternative Medicine,
60: 59-84. 5(3): 313-316.
2. Banksota, A.H., Y. Tezuka and S.H. Kadota, 2001. 14. Righi, A.A., G. Negri and A. Salatino, 2013.
Recent progress in pharmacological research of Comparative chemistry of propolis from eight
propolis. Phytotherapy Research, 15: 561-571. Brazilian localities. Evidence-Based Complementary
3. Parolia, A., M. Tomas, M. Kundabala and M. Mohan, and Alternative Medicine, http:// dx.doi.org/ 10.1155/
2010. Propolis and its potential uses in oral health. 2013/267878.
Int. J. Med Medical Sci., 2(7): 210-215. 15. Cirasino, L., A. Pisati and F. Fasani, 1987.
4. Salatino, A., E.W. Teixeira, G. Negri and D. Message, Contact dermatitis from propolis. Contact Dermatitis,
2005. Origin and chemical variation of Brazilian 16: 110-111.
propolis. Evidence-Based Complementary and 16. Almas, K., A. Dahlan and A. Mahmoud, 2001.
Alternative Medicine, 2(1): 33-38. Propolis as a natural remedy: An update. Saudi
5. Simone-Finstrom, M. and M. Spivak, 2010. Dental J., 13: 45-49.
Propolis and bee health: the natural history and 17. Park, Y.K., S.M. Alencar and C.L. Aguiar, 2002.
significance of resin use by honey bees. Apidologie, Botanical origin and chemical composition of
41: 295-311. Brazilian propolis. J. Agric. Food Chem.,
6. Hegazi, A.G., Z.N. Moharm, F. Abd Allah, E.A.B. El 50: 2502-2506.
Sheikh, N.A. Ellithy and A.M. Khair, 1998. Influence 18. Sforcin, J.M. and V. Bankova, 2011. Propolis: Is there
of ethanolic extract of Egyptian propolis in mice a potential for the development of drugs? J.
bearing Ehrlich carcinoma on immune status. J. Union Ethnopharmacol, 133: 253-260.
of Arab Biologists, 10(A): 387-396. 19. Marcucci, M.C., 1995. Propolis: chemical
7. Makashvili, Z.A., 1978. From the history of propolis. composition, biological properties and the
In Remarkable hive product: Propolis. Scientific data therapeutic activity. Apidologie, 26: 83-99.
and suggestions concerning its composition, 20. Markham, K.R., K.A. Mitchell, A.L. Wilkins,
properties and possible use in therapeutics. J.A. Daldy and Y. Lu, 1996. HPLC and GCMS
APIMONDIA standing commission on beekeeping identification of the major organic constituents in
technology and equipment, Bucharest. New Zealand propolis. Phytochemistry, 42: 205-211.
8. Lotfy, M., 2006. Biological Activity of Bee Propolis 21. Bankova, V., S. De Castro and M. Marcucci, 2000.
in Health and Disease. Asian Pac. J. Cancer Prev., Propolis: Recent advances in chemistry and plant
7: 22-31. origin. Apidologie, 31: 3-15.
9. Popova, M., V. Bankova, S. Bogdanov, I. Tsvetkova, 22. Chen, Y. W., S.W. Wu, K.K. Ho, S.B. Lin, C.Y. Huang
C. Naydenski, G.L. Marcazzan and A.G. Sabatini, and C.N. Chen, 2008. Characterization of Taiwanese
2007. Chemical characteristics of poplar type propolis collected from different locations and
propolis of different geographic origin. Apidologie, seasons. J. Sci. Food Agric., 88: 412-419
38: 306-311. 23. Popova, M., C.N. Chen, P.Y. Chen, C.Y. Huang and
10. Bankova, V., 2009. Chemical diversity of propolis V. Bankova, 2010a. A validated spectrophotometric
makes it a valuable source of new biologically active method for quantification of prenylated flavanones in
compounds. J. ApiProduct and ApiMedical Sci., Pacific propolis from Taiwan. Phytochem. Anal.,
1(2): 23-28. 21: 186-191.

1122
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

24. Popova, M.P., K. Graikou, I. Chinou and 36. Nanasombat, S. and P. Lohasupthawee, 2005.
V.S. Bankova, 2010b. GCMS profiling of diterpene Antibacterial activity of crude ethanolic extracts and
compounds in Mediterranean propolis from essential oils of spices against salmonellae and other
Greece. Journal of Agricultural and Food Chemistry, enterobacteria. KMITL Sci. Tech. J., 5: 527-38.
58(5): 3167-3176. 37. Norajit, K., N. Laohakunjit and O. Kerdchoechuen,
25. Rushdi, A.I., N. Adgaba, N.I.M. Bayaqoob, A. Al- 2007. Antibacterial effect of five Zingiberaceae
Khazim, B.R.T. Simoneit, A.H. El-Mubarak and essential oils. Molecules, 12: 2047-60.
K.F. Al-Mutlaq, 2014. Characteristics and chemical
38. Ozan, F., H. Ozer, H. Ozdemir, K. Eren and H. Yeler,
compositions of propolis from Ethiopia. SpringerPlus
2008. A morphometric and histopathologic evaluation
2014, 3: 253.
of the effects of propolis on alveolar bone loss in
26. Abouzid, M.M. and A.A. Abd El-Kaway, 2013.
experimental periodontitis in rats. J. Periodontol.,
Chemical composition of Egyptian and commercial
propolis and its effects on liver function and lipid 79: 1089-94.
profiles in albino rats. J. Biological Chem. Env. Sci., 39. Allen, K.L., P.C. Molan and G.M. Reid. 1991. A
8(2): 323-340. survey of the antibacterial activity of some New
27. Hegazi, A.G., K. Faten and F.K. Abd El Hady, 1997. Zealand honeys. J. Pharm. Pharmacol., 43: 817-22.
Chemical and biological studies of Egyptian propolis. 40. National Committee for Clinical Laboratory
International Symposium on Apitherapy, Cairo 8-9 th Standards, 2000. Methods for dilution antimicrobial
March. susceptibility tests for bacteria that grow
28. Kimoto, T., S. Arai, M. Kohguchi, M. Aga, anaerobically. Approved Standard. (4th ed.), 20(2) M7-
Y. Nomura, M.J. Micallef, M. Kurimoto and K. Mito, A5.
1998. Apoptosis and suppression of tumor growth by 41. Salonen, A., S. Saarnio and R. Julkunen- Tiitto, 2012.
artepillin C extracted from Brazilian propolis. Cancer Phenolic compounds of propolis from the boreal
Detection and Prevention, 22: 506-15. coniferous zone. Journal of Apicultural Science,
29. Hegazi, A.G. and F.K. Abd El Hady, 2002. Egyptian 56(1): 13-22.
propolis: 3. Antioxidant, antimicrobial activities and
42. Hegazi, A.G., A.M. Abdou and F. Abd Allah, 2014.
chemical composition of propolis from reclaimed
Egyptian Propolis 11: Its antimicrobial activity with
lands, Z. Naturforsch., 57c: 395-402.
comparison with different localities. Int. J. Curr.
30. El Sohaimy, S.A., 2014. Chemical Composition,
Antioxidant and Antimicrobial Potential of Artichoke. Microbiol. App.Sci., 3(9): 530-538.
The Open Nutraceuticals Journal, 7: 15-20. 43. Ramanauskien, K., A.M. Inknien, V. Petrikait and
31. Wang, H., X.D. Gao, G.C. Zhou, L. Cai and W.B. Yao, V. Briedis, 2013. Total phenolic content and
2008. In vitro and in vivo antioxidant activity of antimicrobial activity of different Lithuanian propolis
aqueous extract from Choerospondias axillaris fruit. solutions. Hindawi Publishing Corporation,
Food Chem., 106: 888-895. Evidence-Based Complementary and Alternative
32. Barbary, O.M., S.A. El-Sohaimy, M.A. El-Saadani and Medicine, volume 2013: 2-5.
A.M.A. Zeitoun, 2010. Antioxidant, antimicrobial and 44. Yang, H., Y. Dong, H. Du, H. Shi, Y. Peng
anti-HCV activities of lignan extracted from flaxseed. and X. Li, 2011. Antioxidant compounds from
Research Journal of Agriculture and Biological propolis collected in Anhui, China. Molecules, 16:
Sciences, 6(3): 247-256. 3444-3455.
33. Croci, A.N., B. Cioroui, D. Lazar, A. Corciova, 45. Abd El-Hady, F.K. and A.G. Hegazi, 1994, Gas
B. Ivanoscu and M. I. Lazar, 2009. HPLC evaluation chromatography-mass spectrometry (GC/MS). Study
of phenolic and polyphenolic acids from propolis.
of the Egyptian propolis 1- aliphatic, phenolic acids
Farmacia, Vol. LVII(1): 52-57.
and their esters. Egypt. J. Appl.Sci., 9: 749-760.
34. Julkunen-Tiitto, R. and S. Sorsa, 2001. Testing the
46. Abd El-Hadya, F.K., A.G. Hegazi and
drying methods for willow flavonoids, tannins and
E. Wollenweber, 2007. Effect of Egyptian propolis
salicylates. J. Chem. Ecol., 27(4): 779-789.
35. Bauer, A.W., W.M.M. Kirby, J.C. Sherris and on the susceptibility of LDL to oxidative
M. Turck, 1966. Antibiotic susceptibility testing by a modification and its antiviral activity with special
standardized single disk method. The American emphasis on chemical composition. Z. Naturforsch.,
Journal of Clinical Pathology, 45: 493-496. 62c: 645-655.

1123
Am-Euras. J. Agric. & Environ. Sci., 14 (10): 1116-1124, 2014

47. Hegazi, A.G., A.A. Farghaly and F.K. Abd El Hady, 52. Abd El Hady, F.K. and A.G. Hegazi, 2001. Egyptian
2001. Antiviral activity and chemical composition of propolis: 2- chemical composition, antiviral and
European and Egyptian propolis. Proc. 37th Int. Apic. antimicrobial activities of East Nile delta propolis.
Congr. 28 Oct - 1 Nov 2001, Durban, South Africa. Proc. 37th Int. Apic. Congr. 28 Oct - 1 Nov 2001,
48. Sonmez, S., L. Kirilmaz, M. Yucesoy, B. Ycel and Durban, South Africa.
B. Yilmaz, 2005. The effect of bee propolis on oral 53. Cowan, M.M., 1999. Plant products as antimicrobial
pathogens and human gingival fibroblasts. J. agents. Clin. Microbiol. Rev., 12: 564-82.
Ethnopharmacol., 102: 371-6. 54. Rao, A., Y. Zhang, S. Muend and R. Rao, 2010.
49. Gebara, E.C, L.A. Lima and M.P. Mayer, 2002. Mechanism of antifungal activity of terpenoid
Propolis antimicrobial activity against phenols resembles calcium stress and inhibition of
periodontopathic bacteria. Braz. J. Microbiol., the TOR pathway. Antimicrob. Agents Chemother,
33: 365-9. 54(12): 5062-9.
50. Santos, F.A., E.M. Bastos, M. Uzeda, M.A. Carvalho, 55. Ahmad, A., A. Khan, F. Akhtar, S. Yousuf, I. Xess,
L.M. Farias, E.S. Moreira and F.C. Braga, 2002. L.A. Khan, N. Manzoor, 2011. Fungicidal activity of
Antibacterial activity of Brazilian propolis and thymol and carvacrol by disrupting ergosterol
fractions against oral anaerobic bacteria. J. biosynthesis and membrane integrity against
Ethnopharmacol., 80: 1-7. Candida. Eur. J. Clin. Microbiol. Infect. Dis.,
51. Agarwal, G., G.G. Vemanaradhya and S.M. Dhoom, 30(1): 41-50.
2012. Evaluation of chemical composition and
efficacy of Chinese propolis extract on
Porphyromonas gingivalis and Aggregatibacter
actinomycetemcomitans: An in vitro study. Contemp
Clin Dent., 3(3): 256-261.

1124