Antioxidative activity of propolis from Dalmatia (Croatia)

Katalinić V, Radić S, Ropac D, Mulić R, Katalinić A.
Odjel sanitarne kemije i toksikologije, Institut pomorske medicine HRM, Split, Hrvatska.

AIM: The aim of this study was to determine the antioxidative activity of propolis from ecologically clean parts of Dalmatia.

METHODS: Phenol concentration in ethanolic propolis extracts was determined by Folin-Ciocalteu reagent using gallic acid as the standard. Flavonoid phenolic compounds were analyzed after precipitation with formaldehyde. The residual non-flavonoid phenolics were also determined by Folin-Ciocalteu method. By determining the change of peroxide number (PN), of tiobarbiture acid reactive species (TBARS), and of DPPH-radical activity, antioxidative efficiency of propolis was tested and compared with well known and widely used synthetic antioxidants. Values of PN and TBARS were determined at 60 degrees C in samples of trigyceride substrate (lard) without and with the addition of antioxidants. Compared was the efficiency of three antioxidants: propolis (alcoholic extract), vitamin E, and (+)-catechin in a concentration of 1%. PN was monitored during 50 days. By the method of Sedlacek, TBARS were measured during 30 days. Antioxidative activity of propolis extract was also measured in terms of hydrogen donating ability using stable radical alpha,alpha-diphenyl-beta-picril hidrazyl (DPPH*) and compared with commercial synthetic antioxidants of butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), and (+)-cathecin. Inhibition degree of DPPH* was calculated by the formula of Yen and Duh.

RESULTS: Total phenol content, expressed as gallic acid, in propolis extracts varied from 75.2 to 90.2 g/kg propolis. The proportion of flavonoids in total phenols ranged from 62% to 65%. Values of TBARS were not increased only in samples with added propolis. The inhibition of DPPH-radical by propolis extracts ranged from 93% to 96%, by catechin 95%, by BHT 49%, and by BHA 64%. Compared to BHT and BHA, propolis extracts showed greater reducing activity against DPPH-radical.

DISCUSSION: The chemical composition of propolis, and thus its biological activity depend on the plant from which it has been collected, and on the macro- and microclimatic conditions. Many compounds in propolis exert antioxidative activity. A belief was expressed that the biological activity of propolis is very probably based mostly on its antioxidative efficiency. Dalmatian propolis showed high efficiency in the prevention of oxidative processes. This could be explained by the high proportion of polyphenol constituents, especially flavonoids. A very low and equal degree of increase of PN, as a measure of oxidative processes, was noticed in the samples of triglyceride substrate with the addition of propolis and (+)-catechin. The greatest rise of TBARS was measured in the samples of pure lard. There was no increase of TBARS only in the samples with added propolis. Propolis and (+)-catechin showed great efficiency in the inhibition of DPPH-radical, greater than BHT and BHA, which are widely used in food industry.

CONCLUSION: The results indicate that Dalmatian propolis could be an efficient protective agent against oxidative processes in food. The high antioxidative activity of propolis, its natural origin, and present knowledge about its biological properties, make it a very promising nutritional additive for human diet.

Oxidant/antioxidant properties of Croatian native propolis

Sobočanec S, Šverko V, Balog T, Šaric A, Rusak G, Likić S, Kušić B, Katalinić V, Radić S, Marotti T.

Division of Molecular Medicine, Ruder Boskovic Institute, Zagreb.

Native propolis was defined as propolis powder collected from the continental part of Croatia and prepared according to a patented process that preserves all the propolis natural nutritional and organoleptic qualities.

Nine phenolic compounds (out of thirteen tested) in propolis sample were detected by high performance liquid chromatography (HPLC) analysis. Among them chrysin was the most abundant (2478.5 microg/g propolis). Contrary to moderate antioxidant activity of propolis examined in vitro (ferric reduction antioxidant power; FRAP-assay), propolis as a food supplement modulated antioxidant enzymes (AOE) and significantly decreased lipid peroxidation processes (LPO) in plasma, liver, lungs, and brain of mice. The effect was dose- and tissue-dependent. The lower dose (100 mg/kg bw) protected plasma from oxidation, whereas the higher dose (300 mg/kg bw) was pro-oxidative. Hyperoxia (long-term normobaric 100% oxygen) increased LPO in all three organs tested.

The highest vulnerability to oxidative stress was observed in lungs where hyperoxia was not associated with augmentation of AOE. Propolis protected lungs from hyperoxia by increased catalase (CAT) activity. This is of special importance for lungs since lungs of adult animals are highly vulnerable to oxidative stress because of their inability to augment AOE activity. Because of its strong antioxidant and scavenging abilities, native propolis might be used as a strong plant-based antioxidant effective not only in physiological conditions but also in cases that require prolonged high concentration of oxygen.

PROPOLIS - 21st Century medication?

Propolis lijek 21. stoljeća?

Propolis is a resin collected by bees mostly from flower and leaf buds. Historical monuments indicate that it has been used as a remedy since ancient times. It is known to have a wide spectrum of therapeutic effects.

Wound healing

It assists in wound healing, not only when applied locally, but also when taken by mouth.


Survival of mice after exposure to gamma rays is attributed to propolis’s antioxidative properties and it indicates its potential for protection from free radicals produced by various types of radiation.


Protects the stomach from alcohol damage.


When a stroke is suffered, propolis, in synergy with pollen, strengthens the antioxidative defence and improves blood supply of the brain which results in more rapid rehabilitation of disrupted and lost brain functions.


Propolis prevents cataract development and enhances the antioxidative defence of blood and lenses, thus protecting cornea from the consequences of chemical injuries.

Anti-oxidant properties

Antioksidacijska svojstva

At the present state of knowledge we could say that the basis of biological activity is to be found in propolis’s strong anti-oxidant properties because it inhibits and scavenges free radicals, which are known to cause a number of diseases, such as tumours, cardiovascular diseases, nervous system diseases, cataract, emphysema, premature ageing and others.

Anaesthetic properties

Its local anaesthetic properties are equivalent to those of Lidocaine and greater than those of cocaine and Procaine. Propolis is also useful as an analgesic agent when taken by mouth.

Anti-inflammatory properties

Strong anti-inflammatory effect is visible in cases of acute as well as chronic inflammation. It is interesting that it is also effective against non-infectious inflammations (burns caused by sun, fire, radiation and chemicals).

Antiparasitic properties

It blocks the development of single cell parasite Trichomonas vaginalis, which causes inflammation of female reproductive organs and male urethra. It is also effective against Trypanosoma cruzi.

Antiviral properties

Propolis inhibits the reproduction of a number of viruses that cause human diseases (Herpes simplex, genitalis and zoster, influenza and smallpox).

Clinical tests have showed that the time of recovery from common cold can be reduced by 2,5 times when propolis is used.

Antifungal properties

It is effective against various types of fungi, mainly those causing skin and hair diseases.

Antimicrobial properties

Biološka svojstva

Propolis has a stronger antibacterial effect on gram-positive bacteria (streptococcus, staphylococcus) than on gram-negative bacteria (Salmonella, Escherichia coli, Proteus spp., Pseudomonas aeruginosa).

Furthermore, it is beneficial in treatment of tuberculosis in human patients. It is interesting to note that, even when it is not directly effective against gram-negative bacteria, propolis creates protection against inflammation caused by those bacteria (Klebsiella pneumoniae, Proteus vulgaris, Pseudomonas aeruginosa).

The effect is less strong as well as less toxic than that of antibiotics, it does not cause bacteria resistance and it does not damage healthy intestinal flora.

When used in synergy with some of the antibiotics, the time of recovery is significantly reduced.

Physical and chemical characteristics of propolis

Terapijska svojstva

Propolis is a resinous substance of a yellowy green to brown or dark red colour, a balsamic aroma, and a bitter taste. To this day, over 300 biologically active substances have been isolated from propolis.

The most important among them are flavonoids, various phenolics and aromatic compounds.

The use of propolis in its original form is becoming increasingly difficult nowadays, due to pollution in areas where bees collect it. Thus, it needs to be chemically treated before it can be used by humans.

These procedures are a cause of partial loss of biologically active substances. Therefore, propolis solutions do not retain all the compounds originally contained in propolis since it is not completely soluble in any known solvent.

Due to harmful effects of solvents, it must not be used in larger quantities and large dosages are prone to cause stomach pains.

Company HEDERA had succeeded in producing native propolis in its original form without the use of chemical solvents, using its innovative methods. It has been collected in ecologically clean parts of Croatia.

This is a dietetic product of the highest purity, containing all biologically active substances. It can be used in larger dosages without harmful side effects because it does not contain any solvents.

There is no similar product on the market.




Learn more about the workings of propolis on the organ. Click on the image for visual display.

The influence of propolis on human body



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