1

HONEYBEE PROPOLIS: A NATURAL RESOURCE OF

BIOACTIVE COMPOUNDS

(A BRIEF REVIEW)

1.1 LITERATURE REVIEW

Propolis (Cas No. 9009-62-5), also referred as bee-glue is sticky dark coloured

complex mixture of compounds. For producing propolis honeybees collect

phytochemicals from surrounding flora, add their salivary enzymes followed by

addition of wax produced by them (Marcio et al, 2011: 208-219). Honeybees fill

cracks and crevices in the brood frames with propolis. It is also used to reduce the

size of the entrance of the bee hive. Probably this application has led to the name

“propolis” (Greek pro- before, police- city). Honeybees are also known to use

propolis for embalming the carcass of a hive invader which has been killed but

cannot be transported out of the hive (Singh, 1972: 11-19). It is, thus, a substance

that is used for maintenance and defense of the hive. When honey is extracted from

the colony, propolis is also obtained along with wax and pollen grains. Thus it is

regarded as one of the beehive products. Propolis, when obtained from the bee hive,

is usually yellow to dark reddish brown in colour (Fig. 1.1).

Figure 1.1 Propolis collected from honeybee hive

From a beehive around 200 g of propolis is uaually obtained at a time. It possesses a

peculiar odour and is brittle. It has comparatively low melting point which is in the

2

range 65ºC to 75ºC (Ghisalberti, 1978: 59-84). Considering its availability in

abundance, its applications and composition were studied since the beginning of

19th century. A large variation in the composition from these studies shows gross

composition of propolis determined in early years (Table 1.1).

Table 1.1 Gross composition of propolis reported in early years

Resin +

Wax (%)

Balsam

(%)

Volatiles

(%)

Alcohol

soluble

part (%)

Residue

(%)

Reference

80.3 - 0 6.7 12.9 Helfenberg, 1908: 987-988

8.7 + 27.9 - 6.9 - 12.9 Bohrisch, 1908: 927-929

64.6 + 16.1 - 6 12.9 Some Dieterich, 1908: 369

78.6 + 72.6 3 - 8 - - 13.4 Dieterich, 1911: 55771

60 11 6 - 22 Dieterich, 1911: 1019-

1027

70.7 +14.8 5 4.2 - 5.3 Heiduschka & Vogel,

1913: 1087-1988

70 + 30 - - - - Caillas A 1923: 1145-1147

68.9 + 19.3 4 0.5 - 4.3 Jungkunz R 1932: 30-33

50 + 40 - 10 - - Nicolas A 1947: 142-143

Besides studying composition, applications of propolis and its fractions have also

been investigated.

Examination of literature reveals a large range of references about the

biological evaluation of propolis and its fractions. The properties of propolis have

attracted the attention of scientist since the late 60s. During the last 50 years many

investigations have been published on the chemical composition, pharmacology and

therapeutical uses of propolis (Ghisalberti 1978: 69-84). Philip (1928: 705-714) was

one of the earliest to suspect its antiseptic quality. In 1960, Lavie (1960: 103-183,

201-305) found that Propolis showed bacteriostatic activity. Lindenfelser (1967: 90-

92,130-131) reviewed the antiseptic properties of propolis, which have been used in

3

antiseptic preparation for treatment of surgical cases. Table1.2 summarizes the

different activities shown by the propolis from different countries

Table 1.2 Activities proved for propolis from different regions

Activity Source Reference

Antibacterial

Russia Vachonina et al, 1969: 185; Shevenko et al,

1969: 45; Glinnik et al, 1981: 75-76

Romania Ioirich et al, 1964: 191-200

Poland Scheller et al, 1968: 391-398 ; Kedzia et al,

1986 : 187-95 ; Meresta et al,1985: 61-63

France Kivalkina et al, 1969 : 103-105 ; Bakay et al,

1975 : 225-227 ; Barre, 1942 : 584-490

England Alfansus, 1933 : 92-93, Aliev, 1968: 105

Brazil

Bankova et al, 1995 : 167-172 ; Antunes et

al, 1996 : 15-18 ; Aga et al,1994 : 945-946

Chile Valcic et al, 1999: 406-416

Greek Eleni et al, 2006: 992-996

Iranian Shiva et al, 2006: 327-333

Anatolian Ataç et al, 2005: 69-73

Bulgaria Kujumgiev et al,1993: 785-786

Japan Shimada et al, 2001: 11

Egypt Hady et al, 2003:

Antioxidant

Saradian Cengarle et al,1998: 551-557

China, Australia,

New Zealand

Oyaizu et al, 1999a : 135-138 ; 1999b: 135-

138

Canary Island Christov et al,1999: 89-92

China Zhang, 2002 : 48-52 ;Qi et al, 2001 : 11-21

Cuba Pascual et al, 1994 :9-13

Korea Choi et al, 2006 : 29-32

Brazil Bankosta et al, 2000a: 16-23; Hayashi et al,

1999: 1521-1524

4

Activity Source Reference

Taiwan Lu et al, 2006: 277-282

Japan Kumazawa et al, 2001 : , Nagai et al, 2003 :

29-33

Hepatoprotective

Brazil, Peru,

Netherlands

Bankosta et al, 2000a: 16-23

Antiinflammatory

Korean Eun-Hee et al, 1996: 337-341

Romania Oita et al, 1979: 2

Anaesthetic

Brazil Nicolus, 1947: 142-143

Bulgaria Tsakov, 1973:38-41

Antiulcer

Russia Gorbatenko,1971: 22-24

Respiratory Track infections

Russia Aliev, 1968: 105; Kravchuk, 1968: 97-98

Antiviral

France Kravchuk et al, 1968: 97-98

Antitumour

Japan Suzuki et al, 1996: 1-6

Korea Choi et al, 1999: 29-31

Antifungal

Romania Popesco et al,1967: 57-66

Brazil Ota et al, 2001: 9-10

Anticancer

Korean Lee et al, 2000: 54-57

Romania Nistor et al, 1985: 2

Health care food

Japan Ishihara et al,1997 : 4 ;Misono et al, 1997 :

10

5

Considering the large range of activity and also its availability on a wider scale,

number of commercial products got emerged. A few popular products are shown in

Fig. 1.2. Reports on the commercial use of propolis appeared in the early 1900s. A

preparation of propolis and vaseline, 'propolisin vasogen' was shown to possess

antibacterial activity (Mayer et al, 1902: 1414) which found an application as a

medication in the Boer war. Other important commercial products developed from

propolis are listed in Table 1.3

Figure 1.2 Popular commercial products manufactured from propolis

6

Table 1.3 Commercial products from propolis

Sr.

No

Commercial product Country Reference

1 Preparation against mycotic infections Germany Bekenmeier et al, 1973: 443-

449

2 Antibacterial ointment Russia Thikonov and Krivenchuk

1973

3 Mouthwash as a antiseptic Russia Jenko et al, 1952

4 Hair tonic Japan Hirahara, 1997: 4

6 Health food China Sun, 2001: 19

7 Dental gel Brazil Itice,1997: 5

8 Dental cream Brazil Pinto, 1998: 7

9 Ointment for burns Brazil Cota de lima,1998: 4

10 Medicinal soap China Zhang,1999: 4

11 Spray for treating burn Romania Palos et al,1978: 2

12 Sunscreen ointment Romania Palos et al,1975: 2

13 Gel for mucosal infections Romania Oita et al,1979: 2

14 Drug: for the treatment of liver diseases Romania Mihail et al,1986: 3

15 Bandage for ophthalmic use Romania Constantinescu et al,1981: 2

16 Toothpastes with improved anti-

inflammatory properties

USA Sklyar et al,1986

17 Preparation for bone regeneration Romania Gidoiu et al,1986: 2

18 Blood lipid-lowering and regulating

agent in the form of capsule, tablet, or

granule

China Zhang et al, 2004: 4

19 Emulsion used in treatment of wound

healing

Poland Katarzyna and Zbigniew ,

2005: 3

20 Disinfectant China Wang et al, 2005: 10

21 Capsule used in treating AIDS and

improving immunity

China Hu et al, 2005: 4

7

As propolis is derived from many sources, its composition bound to be complex and

variable. Dietrich was the earliest one who detected the traces of vanillin in the

propolis during estimation gross composition of propolis (Dieterich, 1911: 1019-

1027). Sixteen years later Jaubert (1927:1134-1136) found that the colour of the

propolis was due to the flavonoidal pigment like chrysin. No report on identification

of other compounds in propolis appeared until 1969. In 1969 Popravko (1969: 476-

482) isolated six flavonoids from the propolis and identified them as acacetin,

kaempferide, rhamnocitrin, 5-hydroxy 4’,7-dimethoxy flavone, 5,7-dihydroxy 3,4’-

dimethoxy flavone and 3,5-dihydroxy 4’,7’-dimethoxy flavone. Two flavanones:

pinostrobin and 5-hydroxy 4’, 7-dimethoxy flavone and an aldehyde, isovanillin was

also isolated. While these developments were going on, the detailed study of the

chemical constitution of propolis and its fractions was being carried out in many

countries. Their results indicated that the chemical constitution of propolis was not

fixed. It was found to change as per the habitat from which the propolis was

collected. This was expected since certain chemical components collected from the

surrounding flora are known to constitute propolis. Due to this the chemical

constitution of propolis from any particular habitat becomes unique. Table 1.4

illustrates the chemical composition of propolis collected from various countries.

Table 4. Chemical composition of propolis from various countries

Source Compounds identified Reference

Flavonoids

England 3,5,7-Trihydroxy-4’,6-dimethoxyflavone

(Betuletol), 3,5,4’,7-tetrahydroxy-5’-

methoxyflavone (Isorhamnetin), 3,4’,5,7-

tetrahydroxyflavone (Kaempferol), 4’,5,7-

trihydroxyflavone (Apigenin), 3,4’,5-trihydroxy-

5’7-dimethoxyflavone (Rhamnazin), 7-

methylquercetin (Rhamnetin)

Vanhaelen and

Vanhaelen et al,

1979: 253-259

Japan 3,5,7-Trihydroxy-6-methoxyflavone, 5,7-

dihydroxy-3-methoxyflavone, 7-hydroxy-5-

methoxyflavanone (Alpinetin), 3,5,7-trihydroxy

Machida et al,

1985: 455-460

8

6-methoxyflavanone, 2,4’,6-trihydroxy-4-

methoxychalcone

Russia 4’,5,7-Trihydroxy-6-methoxyflavone, 5,7-

dihydroxy-3,4’,6-trimethoxyflavone, 3,4’,5,7-

Tetrahydroxy-3’-methoxyflavone, 3,4’,5-

trihydroxy-3’7-dimethoxyflavone

Popravko et al,

1976: 38-41

Poland 2,6-Dihydroxy-4-methoxychalcone, tectochrysin,

pinocembrin, chrysin, galangin, genkwanin,

apigenin, kaempferol, pilloin, 5-hydroxy-4', 7-

dimethoxyflavone, and pinostrobin chalcone.

Pilloin and pinostrobin chalcone

Maciejevicz et al,

1985 :23-25 ;

Maciejevicz et al,

2001 : 1171-1179

Tunasia Myricetin 3,7,4’,5’-tetramethyl ether, Quercetin

3,7,4’,5’-trimethyl ether

Martos et al, 1997:

2824-2829

Brazil Aromadendrine-4’methyl ether, 3,5,7-trihydroxy-

6,4’-dimethoxyflavone

Banskota et al,

1998: 896-900;

Boudourova et al,

1997: 676-679

China Genistein, kaempferol, acacetin and chrysin Volpi et al, 2006:

354-361

Japan Isonymphaeol-B, nymphaeol-A, nymphaeol-B,

and nymphaeol-C

Kumazawa et al,

2004: 260-262

Venezuela Eupatorin, hispidulin, 5-hydroxy-6,7,3',4'-

tetramethoxyflavone

Tomas-Barberan et

al, 1993: 191-6

Beijing 5,7-Dihydroxyflavone, 3,5,7-trihydroxyflavone,

5,7-dihydroxy-2'-methoxyflavone,

Wang et al, 1988:

613-14

Czechoslovakia Pinostrobin (I), pinocembrin (II) Suchy et al, 1981:

543-8

Australlia Pinostrobin (I) , sakuranetin (II) , isosakuranetin

(III, xanthorrhoeol (IV) , pterostilbene (V) , 5-

hydroxy-4'7-dimethoxyflavanone, 3,5-

dimethoxybenzyl alcohol

Ghisalbert et al,

1977: 111-30

9

Source Compounds identified Reference

Acids and Phenols

Venezuela Sesamin, aschantin, sesartenin andyangambin Trusheva et al,

2004: 683-689

Brazil Caffeoylquinic acid derivatives ,C-

guayacylglycerol ,3-[ 4-hydroxy- 3- (3- oxobut-1-

enyl)-phenyl] acrylic acid , 3,5-diprenyl-4-

hydroxycinnamic acid, 3-prenyl-4-dihydro-

cinnamoyloxycinnamic acid, 2,2,-dimethyl-6-

carboxyethenyl-2H-1-benzopyran, 9-E-,2-

dimethyl-6-carboxyethenyl-8-prenyl-2H-1-

benzopyran, 3-prenyl-4-hydroxycinnamic acid,

3prenyl-4-(2-methoxypropionyl)-cinnamic acid,

(E)-3-[2,3-dihydro-2-(1-hydroxy-1-methylethyl)-

prenyl-benzofuran-5-yl]-2-propenoic acid, 3-

Caffeoylquinic acid, 4- Caffeoylquinic acid

5- Caffeoylquinic acid, 3,5- Caffeoylquinic acid,

4,5- Caffeoylquinic acid, 4,5- Caffeoylquinic acid

metyl ester, 3,4- dicaffeoylquinic acid, 3,4-

dicaffeoylquinic acid methyl ester, new p-

coumaric acid derivatives, one lignin, cinnamic

acid derivatives, (E)-2,3-dihydroconiferyl p-

coumarate, (E)-3-{2,3-dihydro-2-[2-[(E)-p-

coumaroyloxy]-1-methylethyl]-5-benzofuranyl}-

2-propenoic acid, (E)-4-(2,3-

dihydrocinnamoyloxy)cinnamic acid, (E)-3-(2,2-

dimethyl-3,4-dihydro-3-hydroxy-2H-1-

benzopyran-6-yl)-2-propenoic acid, (E)-3-[2,3-

dihydro-2-(1-methylethenyl)-5-benzofuranyl]-2-

propenoic acid, (E)-3-[2,3-dihydro-2-(1-

methylethenyl)-7-prenyl-5-benzofuranyl]-2-

propenoic acid and (E)-3-{3-[(E)-4-(2,3-

dihydrocinnamoyloxy)-3-methyl-2-butenyl]-4-

Basnet et al, 1996 :

1479-1484 ;

Tatefuji et al,

2004 : 966-970 ;

Tazawa et al,

2004 : 1477-1479 ;

Basnet et al, 1997 :

828-833

10

hydroxy-5-prenylphenyl}-2-propenoic acid, 4-

hydroxy-3-prenylbenzoic acid

England Salicylic acid (2-hydroxybenzoic acid), 4-

hydroxybenzoic acid, 4-methoxybenzoic acid,

Gentisic acid (2,5-dihydroxybenzoic acid),

Protocatechuic acid (3,4-dihydroxybenzoic acid),

Gallic acid (3,4,5-trihydroxybenzoic acid), ο-

Coumaric acid, p-Coumaric acid, Benzyl ester of

p-coumaric acid, Ferulic acid, Isoferulic acid

Coumarin, Scopoletol

Vanhaelen and

Vanhaelen et al,

1979: 253-259

Japan Cinnamylidene acetic acid

3,5-dihydroxystilbene (pinosylvin)

Machida et al,

1985: 455-460

Poland 2-Amino-3-methoxybenzoic acid,

Phenylmethyl ester of benzoic acid,

Phenylmethyl ester of salicylic acid,

3,4-dimethoxycinnamic acid, Cinnamic acid

methyl ester, Cinnamic acid ethyl ester, 1,5-

pentanediol monobenzoate, 6,10,14-trimethyl-2-

pentadecanone, 2-heptadecanone

1-(4-Hydroxy-3-methoxyphenyl)1,2-bis{4-[(E)-

3-acetoxypropen-1-yl]-2-methoxyphenoxy}-

propan-3-ol acetate, 1-(4-hydroxy-3-

methoxyphenyl)-2- {4-[(E)-3-acetoxypropen-1-

yl]-2- ethoxyphenoxy}propan-1,3-diol 3-acetate,

3-acetoxymethyl-5-[(E)-2-formylethen-1-yl]-2-

(4-hydroxy-3-methoxyphenyl)-7-methoxy-2,3-

dihydrobenzofuran

Maciejevicz et al,

1983 : 251-253 ;

Maciejevic et al,

1984 : 421-427 ;

Valcic et al, 1998:

95-110

USA Caffeic acid, ferulic acid

Johnson et al,

1994 : 1783-91

Korea Gallic acid, chlorogenic acid, catechin,

epigallocatechin gallate, epicatechin, epicatechin

gallate

Jeong et al, 2003:

501-505

Bulgeria p-Coumaric acid, dimethoxycinnamic acid, Bankova et al,

11

isoferulic acid, ferulic acid, caffeic acid, pentenyl

ferulate, pentenyl caffeate, pinocembrin,

pinobauksin, pinobanksin-3-O-acetate benzyl

caffeate, galavgin

1989: 94-8

Terpenoids

Poland Geraniol, Nerolidol, Guaiol, Farnesol

Dihydroeudesmol, β-bourbonene, Caryophyllene,

Patchoulane, Selenene

Aromadendrene, Calarene, Copaene

Calamenene, β-patchoulene, β-bisabolene

Maciejevicz et al,

1983: 251-253

Brazil Communic acid , Imbricatoloic acid Isocupressic

acid Acetyl isocupressic acid, 8 (17),13 E-

labdadien-15,19-dioic acid, 13 E-labdadien-15-

dioic acid, 13-hydroxy-8(17),14- labdadien-19-

dioic acid, β and amyrin, Cycloartenol,

Pentacyclic triterpenoid bauer-7-en-3-yl acetate,

cycloartenol, lanosta-7, 24-diene-3-ol lupeol

Bankova et al,

1996: 277-280

; Banskota et al,

1998: 896-900

; Marcucci et al,

1998: 117-119;

Furukawa et al,

2002: 439-440

Croatia α-pinene, β-pinene, γ-terpinene Borcic et al, 1996:

311-313

Canary Island Geraniol, Linalyl propionate Bankova et al 1998:

1411-1415

Steroids

Poland Cholestrilene, Cholinasterol, Stigmasterol, β-

dihydrofucosterol

Maciejevicz et al,

1982: 277-279

Hydrocarbons

Brazil C27H56, C29H60, C31H64, C33H68.

Carboxylic acids: C16:0, C18:0, C18:1.;primary

alcohols. range from C24 to C34, C30

Negri et al, 1998 :

305-314

Poland Eicosane, Eicosine, Heneicosane1-octadecene,

Tricosane, Pentacosane

Maciejevicz et al,

1982 : 277-279 ;

Maciejevicz et al,

1983 : 251-253

12

Source Compounds identified Reference

Volatiles

Brazil Ledol, spatulenol, germacrene, guaiol, farnesol,

dihydroeudesmol, and nerolidol. -Bisabolene,

patchoulane, Me 2,8-dimethylundecanoate, and

benzyl benzoate

Maciejevicz et al,

1983: 251-253

Greece junipene , α-pinene , manoyl oxide, trans-β-

terpineol α-eudesmol, n-decanal , guaiol, δ-

cadinene , α-muurolene, n-decanal, cedrol, n-

nonanal, manool

Eleni et al, 2006;

992-996

13

Structures of a few important identified compounds from propolis are indicated in

Fig.1.3a and 1.3b.

Figure 1.3a Structures of some important identified compounds from propolis

14

Figure 1.3b Structures of some important identified compounds from propolis

The propolis is thus a natural product whose composition is extraordinary variable.

Samples from different geographic origin may possess totally different chemical

compositions and biological activities. This information in literature not only

illustrates the chemical constitution and applications of propolis from various

countries but it also throws light on the lack of study on Indian propolis. Thus,

15

although a lot of investigation is done on propolis from other countries much less

efforts are made to investigate the biological properties and chemical constitution of

Indian propolis.

1.2 INDIAN SCENARIO

Honey is a common household product and bee-keeping is a well-managed activity

in India. Honey and wax are the main commercial products in India. Other beehive

products like propolis are less known. Though Indian bee-keeping indutries do

produce propolis, its chemistry and applications have not been much investigated.

The work on Indian propolis started in late 90’s. Initially using various

different parameters and models for hepatoprotective activities, it was shown that

Indian propolis acted as a strong hepatoprotective agent (Table 1.5). Results of the

strong hepatoprotective activity generated interest in the study of other biological

activities of Indian propolis.

Table 1.5 Hepatoprotective studies of Indian propolis

Type of the study Reference

The protective effect of propolis against alcohol carbon

tetrachloride induced hepatoxicity in rats

Sharma et al, 1997:

76-81

The protective effect of propolis was studied in paracetamol

induced acute hepatic necrosis in Wistar rats

Sharma et al, 1998:

27-34

Effect in restoring CCl4 induced istopathological alterations Shukla et al, 2004:

993-997

Hepatoprotective efficacy of propolis against biochemical

and histopathological changes induced by CCl4

Bhadauria et al,

2007: 145-54

The curative effect of propolis against acetaminophen

(APAP) induced acute hepatorenal alterations.

Satendra Kumar et al,

2008: 451-461

The duration-dependent hepatoprotective effects of propolis

against carbon tetrachloride induced liver damage in rats

Bhadauria et al, 2007:

1136-1145

Therapeutic potential of propolis as a hepatoprotective agent

after mercury intoxication

Bhadauria et al, 2008:

311-321

The protective effects of propolis on inorganic mercury

induced oxidative stress in mice.

Zhao et al,

2009 :164-269

16

Antimicrobial and antioxidant activity of propolis from Gujarat (Kumar N et al

2008) and south India (Ranjith Kumar et al, 2011: 8-13) were investigated and

reported. In a recent report Indian propolis was proved to be a strong antioxidant. On

the basis of the results it is possible that Indian propolis could be of value as an

easily accessible source of natural antioxidant, as a food supplement or in the

pharmaceutical industry (Naik et al, 2009: 110-120). The antiulcer activity was

proved for Indian propolis (Pillai et al, 2010: 21-28). Its wound healing property

was also demonstrated (Pillai et al, 2010: 1198-1206). The propolis has also been

proved to be effective in cancer treatment. The study showed that propolis has a

pharmacological role in treating experimental breast cancer in rats. It has maximum

activity when administered with paclitaxel (Padmavati et al, 2003: 72-77). Though

these reports on the medicinal activities of Indian propolis were seen, very few

references are available on the chemical composition of Indian propolis. To reveal

the information about chemical composition of Indian propolis the GC-MS analysis

of propolis was carried out. This study investigated the composition of ethanolic

extracts of propolis collected from Gujarat. The compounds identified from the

propolis are pinocembrin, chrysin, galangin, caffeic acid, 3- hydroxy-4-

methoxycinnamic acid, 3,4-dimethoxycinnamic acid, p-coumeric acid, benzyl

cinnamate, 4-pentanoic acid, and ferulic acid (Kumar et al, 2009: 46-48). Another

report on the chemical constitution of Indian propolis appeared in 2010 (Laskar et

al, 2010: 233-237) They isolated galangin and pinicenbrin from ethanol extrtact of

Indian propolis.

Thus, the potential of Indian propolis is not completely explored. Its

chemical constitution, which is expected to be unique, is also not studied in

sufficient depth. Considering the vast potential of Indian propolis as a novel

bioresource, its bio-evaluation followed by the chemical investigation is felt

justified.

17

1.3 REFERENCES

Aga H, Shibuya T, Sugimoto T, Kurimoto M, Nakajima S (1994) Isolation

and identification of antimicrobial compounds in Brazilian propolis. Biosci.

Biotechnol. Biochem. 58(5): 945-6.

Alfonsus EC (1933) Some sources of propolis. Glean. Bee Cult. 61: 92-93.

Aliev BA (1968) Utilisation locale du produit des abeilles, propolis en oto-

rhinolaryngologie pratique. Vest. Oto-rino-Iar. 30: 105.

Antunes RMP, Catao RMR, Ceballos BSO (1996) Antimicrobial activity of

propolis. Rev. Bras. Farm. 77: 15-18.

Ataç U, Kadri˙ye S, Özant Ö, Dilşah Ç, Ömür G, Beki˙r S (2005) Chemical

compositions and antimicrobial activities of four different Anatolian propolis

samples. J Ethnopharmacol. 99: 69-73.

Aubert JGF (1926) Origin of the colour of beeswax and the composition

of propolis.] C. r. hebd. Seanc. Acad. Sci., Paris. 184: 1134-1136

Bakay M, Pusztai R, Beladi I (1975) Effect offlavonols on the in vitro

response of chicken lymphocytes to phytohaemagglutinin. pp. 225-227 from

Topics in flavonoid chemistry and biochemistry. eds Farkas L, Gabor M and

Kallay F. Amsterdam: Elsevier Scientific Publishing Company

Bankova V, Christov R, Delgado Tejera A., (1998) Lignans and other

constituents of peopolis from Canary Island. Phytochemistry 49:1411-1415.

Bankova V, Christov R, Kujumgiev A, Marcucci MC (1995) Chemical

composition and antibacterial activity of Brazillian propolis. Z. Naturforsch.

50c: 167-172.

Bankova V, Kuleva LZ.N (1989) Phenolic compounds of propolis from

different regions of Bulgaria. 26: 94-8.

Bankova V, Marcucci MC, Simova S, Nikolova N, Kujumgiev A, Popov S

(1996) Antibacterial diterpenic acids from Brazillian propolis. Z.

Naturforsch. 51c: 277-280.

Banskota AH, Tezuka Y, Adnyana IK (2000a) Biological evaluation of

propolis from Brazil, Peru, the Netherlands and China. J

Ethnopharmacol. 72: 239-246.

18

Banskota AH, Tezuka Y, Prasain JK, Matsushige K, Saiki I, Kadita Sh

(1998) Chemical constituents of Brazillian propolis and their cytotoxic

activity. J. Nat. Prod. 61: 896-900.

Banskota AH, Tezuka Y, Prasain JK, Matsushige K, Saiki I, Kadita SJ

(1998) Chemical constituents of Brazillian propolis and their cytotoxic

activity. Nat. Prod. 61: 896-900.

Barre R (1942) Identification of the colouring matter of beeswax. Revue.

Can. BioI. 1: 485-490.

Basnet P, Matsuno T, Neidlein R, Naturforsch C (1997) Potent free radical

scavenging activity of propol isolated from Brazilian propolis. Z.

Naturforsch. C: Biosci. 52: 828-833.

Basnet P, matsushige K, hase K, kadota S (1996) Potent antihepatotoxic

acivity of dicafeoyl quinic acids from propolis. Biol. Pharm. Bull. 19: 1479-

1484.

Bekenmeier H, Braun W, Friedrich E, Kala H, Metzner J, Schneidewind E,

Schwaiberger R, Wozniak KD (1973) Mikrobiologische, pharmakologische

und klinische Untersuchungen zur Wirksamkeit von Propolis. Derm. Mschr.

159: 443-449.

Bhadauria M , Satendra Kumar N, Shukla S. (2007) Duration-dependent

hepatoprotective effects of propolis extract against carbon tetrachloride-

induced acute liver damage in rats. Advances in Therapy. 24I1136-1145.

Bhadauria M , Satendra Kumar N, Shukla S (2007) Hepatoprotective

Efficacy of Propolis Extract: A Biochemical and Histopathological

Approach. Iranian. J. Pharmacol. Therapeut. 6:145-154.

Bhadauria M, Shukla S, Mathur M, Agrawal OP, Shrivastava S, Johri S,

Joshi D , Sing V, Mittal D, Satendra kumar N(2008) Hepatic endogenous

defense potential of propolis after mercury intoxication. Integrative zoology.

3: 311-321.

Bohrisch P (1908) Propolis and its characteristic constituents. Pharm.

Zentralhalle. 48: 929-934.

Borcic I, Radonic A, Grzunov K (1996) Comparision of volatile constituents

of propolis gathered in different regions of Croatia. Flav. Fragrance.

J.11:311-313.

19

Boudourova- Krasteva G, Naturforsch Z (1997) Phenolics from Brazilian

propolis. 52c: 676-679.

Caillas A (1923) Composition of the propolis of bees. C. r. hebd. Séanc.

Acad. Sci. Paris. 177: 1145-1147.

Cengarle L, Carta A, Tilloca G, Marceddu MF (1998) Antioxidant activity of

a Sardinian propolis. Riv. Ital. 75: 551-557.

Choi YM, Noh DO, Cho SY, Suh HJ, Kim KM, Kim JM (2006) Antioxidant

and antimicrobial activities of propolis from several regions of Korea. Food

Sci. Technol. 39: 756-761.

Christov R, Bankova V, Tsvetkova I, Kujumgiev A, Tejera AD (1999)

Antibacterial furofuran lignans from Canary Islands propolis.

Fitoterapia 70: 89-92.

Constantinescu M, Dumitrescu A, Ilies N, Trandafir V (1984) Biologically

active ophthalmological bandage. Rom. RO 83236 B, 2 pp.

Cota de lima M (1998) Ointment for treatment of burns. Braz. Pedido PI BR

9603703 A 26 May 1998,4 pp.

Dieterich K (1908) The analysis of bee wax in its several stages of formation

and concerning the bee resin (propolis). Pharm. Post. 40: 369.

Dieterich K (1911) Further contributions to the knowledge of bee resin

(propolis). Pharm. Zentralhalle. 52: 1019-1027.

Dieterich K (1911) Resin from bee glue (propolis). Pharm. Ztg. 55771.

Eleni M, Eleftherios S, Ioanna C (2006) Volatile constituents of propolis

from various regions of Greece – Antimicrobial activity. J. Food

Engineering. 77: 992-996.

Furukawa S, Takagi N, Ikeda T, Ono M, Nafady AM, Nohara T, Sugimoto

H, Doi S, Yamada H (2002) Two novel long-chain alkanoic acid esters of

lupeol from Alecrim-propolis. Chem. Pharm. Bull. 50: 439-440.

Ghisalberti EL (1978) Propolis: A review. Bee Wld. 60: 59-84.

Ghisalberti EL, Jefferies PR, Lanteri R. (1977) Potential drugs from propolis.

Mass Spectrom. Drug Metab. 111-30.

Gidoiu T; Popescu F, Palos E (1986) Preparation for bone regeneration.

Rom. RO 89150 B1, 2 pp.

Glinnik AV, Gapanovich VYa (1981) Antibacterial properties of propolis.

Zh. Ushn., Nos. Gorl. Bolezn. 4: 75-6 (Russian).

20

Gorbatenko AG (1971) Treatment of ulcer patients with a 30% alcohol

solution of propolis. Vrach. Delo. 3: 22-24.

Hady El, Faten KA, Hegazi AG (2002) Egyptian propolis: 2. Chemical

composition, antiviral and antimicrobial activities of East Nile Delta

propolis. Z. Naturforsch. C: J Biosci. 57: 386-394.

Hayashi K, Komura S, Isaji N, Ohishi N, Yagi K (1999) Isolation of

antioxidative compounds from Brazilian propolis: 3,4dihydroxy-5-

prenylcinnamic acid, a novel potent antioxidant. Chem Pharm Bu1l. 47:

1521-1524.

Heiduschka A, Vogel FM (1913) Bee glue (propolis). Pharm. Zentralhalle.

53: 1087-1088.

Helfenberg KD (1908) The analysis of beeswax and propolis.

Chemikerzeitung. 31: 987-988.

Hirahara T (1997) Cosmetics containing propolis-containing compositions.

Jpn. Kokai Tokkyo Koho JP 09067243 A2 Heisei, 4 pp.

Hu GA (2006) capsule for treating AIDS.Faming Zhuanli Shenqing Gongkai

Shuomingshu CN 1771999 A, 4 pp.

Iolrich N, Derevici A, Petrescu A (1965) Antibacterial properties of

Romanian propolis. Lucr. Stllnt. Stat. Cent. Ser. Aplc. 5:107-10.

Ishihara T, Sakamaki K (1997) Propolis dispersion health food. Jpn. Kokai

Tokkyo Koho JP 09149765 A2 10 Jun 1997 Heisei, 4 pp.

Itice NT (1997) Dental gel. Braz. Pedido PI BR 9503177 A, 5 pp.

Jaubert GF (1926) Origin of the colour of beeswax and the composition

of propolis. rhebd. Seanc. Acad. Sci. 184: 1134-1136.

Jenko A (1952) Mouthwash as a antiseptic. Om. Pat. 167862.

Jeong CH, Bae YIl, Lee H, Shim KH (2003) Chemical components of

propolis and its ethanolic extracts. Han'guk Sikp'um Yongyang Kwahak

Hoechi. 32: 501-505.

Johnson KS, Eischen FA, Giannasi DE (1994) Chemical composition of

North American bee propolis and biological activity towards larvae of

greater wax moth (Lepidoptera:Pyralidae). J. Chem. Ecol. 20: 1783-91.

Jungkunz R (1932) Bee’s resin(propolis). Chem.Umschau. 39: 30-33.

Kedzia B, Holderna E (1986) Investigations of the combined action of

antibiotics and propolis on Staphylococcus aureus. Herba. Pol. 32: 187-95.

21

Kedzia B, Holderna-Kedzia E Chemical composition of propolis in up-to-

date studies. Herba. Pol. 1991, 37: 95-110.

Kivalkina VP, Gorshunova VP (1969) Combined action of antibiotics and

propolis on Staphylococcus aureus and Escherichia coli. Fitontsidy, Mater.

Soveshch. 6: 103-105.

Three-component emulsion for treating burns and slowly healing

wounds.Klys, Katarzyna; Klys, Zbigniew (Pol. ). Pol. PL 190340 B1 30 Nov

2005,3pp

Katarzyna K, Zbigniew K (2005) Three-component emulsion for treating

burns and slowly healing wounds. Pol. PL 190340 B1, 3pp

Kravchuk PA (1968) Use of propolis in otolaryngology. Zh. ushn. nosov.

gorlov. Bolez. 28: 97-98.

Kujumgiev A, Bankova V, Ignatova A, Popov S (1993) Antibacterial activity

of propolis, some of its components and their analogs. Pharmazie. 48: 785-6

Kumar N, Mueen Ahmed KK, Dang R, Shivananda TN, Das K (2009) GC-

MS analysis of propolis of Indian origin. Pharmacognosy.1: 46-48.

Kumazawa S, Goto H, Hamasaka T, Fukumoto S, Fujimoto T, Nakayama T

(2004) A new prenylated flavonoid from propolis collected in Okinawa,

Japan. Biosci. Biotechnol. Biochem. 68: 260-262.

Laskar AR, Ismail Sk, Roy N, Naznin AB (2010) Antioxidant activity of

Indian propolis and its chemical constituents Food chem.

Lavie P (1960) Les substances antibactériennes dans la colonie d’abeilles

(Apis mellifica L.) Annls. Abeille. 3: 103-183, 201-305.

Lee SH, Kim DC, Lee JY, Cho M, Hwang WIk (2000) A study on the

anticancer activity of propolis. J. Food Sci. Nutr. 5: 54-57.

Lindenfesler LA (1967) Antimicrobial activity of propolis. Am. Bee. J. 107:

90-92, 130-131.

Lu Li-Chang; Chen, Yue-Wen; Chou, Cheng-Chun (2003) Antibacterial and

DPPH free radical-scavenging activities of the ethanol extract of propolis

collected in Taiwan Yaowu Shipin Fenxi, 11: 277-282.

Machida S, Haga M, Takino Y (1985) Chemical constituents and anti-

microbial activity Japanese propolis. Proc. 30 into Beekeeping Congr. 455-

456.

Maciejewicz W (2001) Isolation of flavonoid aglycones from propolis by a

22

column chromatography method and their identification by GC-MS and TLC

methods. J. Liq. Chromatog. Related Technol. 24: 1171-1179.

Maciejewicz W, Daniewski M, Mielnlczuk Z (1984) Gas chramatagraphy

- mass spectrometry investigation of prapolis. Analysis of phenalic acids

and sugars. Chemia. Ana/it. 2: 421-427.

Maciejewlcz W et al (1983) Gas chromatagraphy - mass spectrometry

propolis. Analysis of sesquiterpenes. Acta Polon. Pharm. 40: 251-253.

Maciejewlcz w. Chemical campositian af propalis: latest investigatian and

research. Internatianal Symposium an Apitherapy, Cracaw, Paland, 23-25

May 1985.

Marcio ARA, Silvana AL, Rosane NMG, Maria MSR, Flavia RFN (2011)

Mechanism of action of underlying antiinflammatory and

immunomodulatory effects of propolis : A brief review. Rev. Brasil. de

Farmacogn. 22: 208-219.

Marcucci MC, Rodriguez J, Ferreres F, Bankova V, Groto R, Popov S

(1998) Chemical composition of Brazillian propolis from Sao Paulo state. Z.

Naturforsch. 53c: 117-119.

Martos I, Cossentini M, Ferreres F, Tomas- Barberan FA (1997) Flavonoid

composition Tunusian honey and propolis. J. Agri. Food. Chem. 54: 2824-

2829.

Meresta L, Meresta T (1986) Antibacterial activity of flavonoid compounds

of propolis, occurring in flora in Poland. Bull. Vet. Inst. Pulawy. 28-29: 61-3.

Meyer W (1902) Neue Arzneimittel und pharmazeutische

spezialitaten. Chem. Ctb. 1414

Mihail N, Nistor C, Makkay C, Nistor V, Onisor M (1986) Drug for

treatment of liver diseases. Rom. RO 88868 B1, 3 pp.

Misono K (1999) Healthy food. Jpn. Kokai Tokkyo Koho JP 11187845 A2

Heisei, 10 pp.

Nagai T, Inoue R, Inoue H, Suzuki N (2003) Preparation and antioxidant

properties of water extract of propolis. Food Chem. 80: 29-33.

Naik, DG; Vaidya, HS; Behera, BC (2009) Antioxidant activity of Indian

propolis. J. ApiProd. ApiMed. Sci. 1:110-120.

Negri G, Marcucci MC, Salatino A, Salatino MLF (1998) Hydrocarbons and

monoesters of propolis waxes from Brazil. Apido. 29:305-314.

23

Nicolas A (1947) Cire d’abeilles et propolis. Nancy: Thomas. 142-143.

Nistor C, Onisor M, Nistor V, Cacoveanu A, Nistor CC (1985) Drug for

treatment of prostate cancer. RO 87149 B1 29 Jun, 2 pp.

Oita N, Palos E, Filipovici (1979) Antiseptic and antiinflammatory

pharmaceutical composition containing bee products. Patent No. RO 69702.

Ota C, Unterkircher C, Fantinato V, Shimizu MT (2001) Antifungal activity

of propolis on different species of Candida. Mycoses. 44: 375-378.

Oyaizu M, Ogihara H, Fujimoto Y (1999a) Antioxidative activity of

extracts from propolis. J. Japn. Oil. Chem. Soc. 48: 135-138.

Oyaizu M, Ogihara H, Fujimoto Y (1999b). Antioxidative activity of

extracts from propolis. Nihon Yukagakkaishi. 48: 135-138.

Padmavati R, Sakethisekaran D (2003) Combined effect of paclitaxel and

propolis against 7,12-dimethyl benz [A] anthracene induced experimental

breast cancer. Biomedicine.23: 72-77.

Palos E, Oita NA (1975) Constanta Sunscreen ointment Rom. RO 59239 30

Sep 1975, 2 pp.

Palos E, Vasilescu VM, Mester I (1978) Spray for treating burns. RO 65190,

2 pp.

Park Eun-Hee; Kim, Sun-Hee; Park, Soo-Sun (1996) Anti-inflammatory

activity of propolis. Arch. Pharmacal. Res. 19: 337-341.

Pascual C, Gonzalez R, Torricella RG (1994) Scavenging action of propolis

extract against oxygen radicals J. Ethnopharmacol. 41: 9-13

Phillip PW. Das K, Seine H, Propolis, its use and origin in the hive. Biol.

Zbl. 1928, 48: 705-714.

Pillai SI, Kandaswamy M, Subramanian S (2010) Antiulcerogenic and ulcer

healing effects of Indian propolis in experimental rat ulcer models J.

ApiProd. ApiMed. Sci. 2: 21-28.

Pillai SI, Palsamy P, Subramanian S, Kandaswamy M (2010) Wound

healing properties of Indian propolis studied on excision wound-induced rats.

J. Pharm. Biol. 48:1198-206

Pinto PS (1998) Prophylactic and therapeutic dental cream propolis

formulation. Braz. Pedido PI BR 9604085 A 16 Jun 7 pp

Popesco A, Braieanu C, Ghlorghlu A, (1967) Antifungal activity of propolis.

Dennato-Vener 12: 57-66.

24

Popravko SA (1976) Plant saurces of propolis. Pchelovodstvo. 96: 38-41.

Popravko SA, Gurevich A, Kolosov MN (1969) Flavonoid components

of propolis. Khimlya Prir. Saedin. 5: 476-482.

Qi Y, Luo S, Liu F, Xu Z, Zheng Y (2001) Antioxidant activity of propolis

Zhongguo Liangyou Xuebao. 16: 18-21.

Ranjith Kumar M, Bose S, Sathyabama S, Brindha P (2011) Antimicrobial

and DPPH Free Radical- Scavenging Activities of theEthanol Extract of

Propolis Collected from India. J. Ecobiotech. 3: 8-13.

Satendra Kumar N, Bhadauria M (2008) Propolis reverses acetaminophen

induced acute hepatorenal alterations: A biochemical and histopathological

approach. Arch. Pharmacal. Res. 31: 451-461.

Scheller S, Rogala D, Stasiak E, Zurek H (1968) Antibacterial properties of

propolis. Polskie Archwm wet. 11: 391-398.

Sharma M, Pillai KK, Husain SZ, Giri DK. (1997) Protective role of

propolis against alcohol-carbontetrachloride induced hepatotoxicity in rats.

Ind. J. Pharmacol. 29: 76-81.

Sharma M, Pillai KK, Hussain Z, Giri DK. (1998) Protective effect of

propolis against paracetamol toxicity in rat liver. Ind. J. Toxicol. 5: 27-34.

Shevenko VS, Shul'man FL (1969) Antimicrobila properties of Russian

propolis. Pchelovodstvo 5: 45

Shevenko VS, Shul'man. FL (1969) Pchelovodstvo. 5: 45.

Shigenori K, Tomoko H, Tsutomu N (2004) Antioxidant activity of propolis

of various geographic origins. Food Chem. 84: 329-339.

Shimada K (2002) Antimicrobial parts containing propolis, their

manufacture, and their application to filters and containers. Jpn. Kokai

Tokkyo Koho JP 2002249411 A2 6 Sep, 11 pp.

Shiva M, Mohammad S, Manoochehr H, Reza A, Nasrin S, Seyed NO

(2006) Chemical composition, oral toxicity and antimicrobial activity of

Iranian Propolis. Microbiol. Res. 161: 327-333.

Shukla S, Bhadauria M, Jadon A (2004) Effect of propolis extract on acute

carbon tetrachloride induced hepatotoxicity. Ind. J. Expt. Biol. 42:993-997.

Singh Z (1972) Propolis collection and its use. Ind. Bee. J. 34: 11-19.

25

Sklyar VE, Tereshina TP, Mozgovaya NV, Gorshkova NV, Ogilets MV,

Danilova AP, Korobka Yu T, Kolobutina VA, Gondel VP, Varava GN

(1986) Toothpastes with improved anti-inflammatory properties U.S.S.R.

SU 1263247 A1

Suchy V, Tekelova D, Petrovic P, Hrochova V, Dolejs L, Farm O (1981)

Study of constituents of propolis of Czechoslovakian origin. I Flavanones

and aromatic acids 50: 543-8.

Sun Q (2001) Health food and its production technology. Faming Zhuanli

Shenqing Gongkai Shuomingshu CN 1322478 A,19 pp.

Suzuki J, Takai H, Koide M, Yamamoto H (1996) Anti-tumor effect of

immunologically active fractions obtained from Brazilian pro polis,

given in combination with anticancer drug in Ehrlich carcinoma bearing

mice. Mitsubachi Kagaku 17: 1-6.

Takeshi N, Reiji I, Hachiro I, Nobutaka S (2003) Preparation and antioxidant

properties of water extract of propolis. Food Chem. 80: 29-33.

Tatefuji T, Izuni N, Ohta T, Arai Sh, ikeda M, kurimoto M (1996) Isolation

and identification of compounds from Brazillian propolis which enhance

microphage spreading and mobility. Biol. Pharm. Bull. 19: 966-970.

Tazawa S, Warashina T, Noro T, Miyase T (1989) Studies on the

constituents of Brazillian Propolis. Chem. Pharm. Bull. 46: 1477-1479.

Tikhonovai, Krivenchuk PE (1973) Preparation of biologically active

poly-phenol&-frem propeHsU.S.$;R. Patent No. 395087.

Tikhonov AI, Krivenchuk PE (1973) Preparation of biologically active

polyphenol &-frem prope Hs. U.S.$;R. Patent No. 395087.

Tomas-Barberan FA, Garcia-Viguera C, Vit-Olivier P, Ferreres F, Tomas-

Lorente F (1993) Phytochemical evidence for the botanical origin of tropical

propolis from Venezuela. Phytochemistry 34: 191-6.

Trusheva B, Popova M, Naydenski H, Tsvetkova I, Gregorio RJ, Bankova V.

(2004) New polyisoprenylated benzophenones from Venezuelan propolis.

Fitoterapia, 75: 683-689.

Tsakov TS, (1973) Antimicrobial properties of Bulgarian propolis. Parmat.

23: 38-41

Vachonina TV, Breeva IG, Dushkova ES (1969) alInd Int. Beekeep. Congr.

Summ, p. 185.

26

Vanhaelen M, Vanhaelen FR (1979) Propolis I. Origine n; tifrographie,

campositiOllr chimique et activite therapeutique. J. Pharm. BelJ. 34: 253-

259.

Volpi N, Bergonzini G (2006) Analysis of flavonoids from propolis by on-

line HPLC-electrospray mass spectrometry. J. Pharmaceut.Biomed. Anal.

42: 354-361.

Wang Bingji, Zhang H (1988) Chemical constituents of Beijing propolis.

Zhongyao Tongbao 13: 613-14.

Wang J, Liu M, Wang Q, Zhou Y, Gan Hui (2005) Preparation of

disinfectant containing nanometer propolis and silver ion. Faming Zhuanli

Shenqing Gongkai Shuomingshu CN 1669434 A 10 pp.

Zhang C (1999) Medicinal soap containing propolis. Faming Zhuanli

Shenqing Gongkai Shuomingshu CN 1221786 A, 4 pp.

Zhang K, Yang R (2004) Blood lipid-lowering and regulating agent. Faming

Zhuanli Shenqing Gongkai Shuomingshu CN 1465399 A, 4 pp.

Zhang Y (2002) Scavenging effect of propolis on free radical. Shipin Yu

Fajiao Gongye. 28: 48-52.

Zhao J-Q, Wan Y-F, Bhadauria M, Satendra Kumar N, Sharma A, Shrivastav

S, Agarwal OP, Mathur R (2009) The protective effects of propolis on

inorganic mercury induced oxidative stress in mice. Ind. J. Expt. Biol. 47:

164-269.