FR2594336A1 - Propolis: purified natural substance employed in agri-foodstuffs, agriculture, cosmetics, dermatology and hygiene products - Google Patents

Abstract

Propolis extract obtained by extraction with alcohol and ultra-pure water. Natural, non-toxic substance allowing the preservation of foodstuffs. Use in agriculture for the destruction of weeds.

Description

 The present invention aims to use a natural substance (propolis) harvested by bees on poplar buds or some conifers. This substance after purification is used in various fields: agribusiness, agriculture, cosmetics, dermatology, hygiene products.

 Propolis, for the part that interests us is composed of vitamins B3 and P.P-. 10% of volatile essential oils, many components of the class of flavonoids, benzoic acid, caffeic, ferulic, chrysin which gives the yellow color of propolis and wax, ceramic alcohol, dimethoxyflavone, dréhéthoxyflavonol , dimethoxyflavonone, vanillin, quercetin, rhamnocitrin, sakuranitrin, galangin, isovaniline, isalfinine, tectochrysin, P benzyl coumarate, pinobanksin, pinocembrin, pinostrobin, pectalinacigenin.

Purified and concentrated propolis contains no lipids, proteins or hormonal substances, but is rich in minerals and trace elements, aluminum, barium, calcium, chromium, cobalt, copper, tin, iron, manganese, nickel, lead, silicon, strontium; titanium, vanadium, zinc, which play a very important role, therefore capital in cellular metabolism.

 The flavonoids mentioned above by their physiological functions, have an interest in therapy by acting directly on blood capillaries, ascorbic acid by potentiating its activity, and by reducing inflammation.

 Propolis, and no publication so far state it among the many works that have been performed on this substance, to say that it could have a teratogenic action, moreover it does not have oncogenic properties in the animal, for example, which as a general rule is perfectly well tolerated and that its use according to the recommended doses and methods leads, except rare exception, no incident or adverse or annoying side effect.

 It has been used for in-vivo testing of doses of 10 and 15 gr per kilo of weight in humans and animals, no toxic effects have been observed or pathological disorder of any kind, even in the long term . Propolis also has extensive bacteriostatic and bactericidal properties on many microbial strains, in particular on certain staphylococci, streptococci, salmonella, bacillus subtilis alvei and larvae, proteus vulgaris, eschérichia coli B (collibacillus).

Fortificidal properties on some species of fungi causing parasitic (fungal) diseases that are related to caffeic acid,
P benzyl coumarate, pino cambrine, pinobaksin.

 Anesthetic properties very powerful, superior to those of cocaine and novocaine that have a relationship with the volatile oils it contains.

 Effective anti-inflammatory properties with healing (stimulates and promotes tissue regeneration).

 Antioxidant properties found by the conservation of certain food extended (2 and 3 times) therefore favorably to be used by the food industry and cosmetics.

 Phytoinhibitory properties by its inhibitory action on the germination of certain plants, thus avoiding in certain cases the irradiation for the germination.

 Propolis also has a particular property, to be mummifying, this power has been found many times, by the presence in the hive of inects, lizards, mice, etc ... which are introduced into it . The bees then cover these unwanted and foreign hosts of propolis, and these after several months of presence remain in a perfect state of conservation and do not emit a putrid odor.

 Propolis natural substance that undergoes no manipulation other than that of the extraction that will be indicated in this file is also devoid of any harmlessness, has no formal counter-indication and incompatibility, it can-therefore be associated without restriction to other therapeutics. It can be used at any age of life without difficulty.

 In view of all these advantages, however, it is necessary to foresee a possible allergic reaction which, moreover, exists and is called apiculturists' dermatosis, but this rare dermatosis is found only at the rate of 0.5 per 1000.

 If in addition a subject is predisposed to the allergy in general, it will be necessary to use the propolis with precaution and especially in those which represent phenomena of cutaneous allergy.

 In some people who are particularly sensitive to this form of cutaneous allergy, even of the eczematous type, which is expressed in the hands by the use often of too large doses in relation to the subject's susceptibility, which has been too frequently used, We will have to put an end to the treatment or the use of propolis, or the benefit of using propolis in progressive doses, which makes it possible to test the sensitivity of the subject.

 Thus propolis by its properties, its quality of natural substance, non-toxic, having no contraindication, is an exceptional product of the first magnitude whose unrestricted use allows its use in many indications.

 Extraction technique to obtain a propolis extract.

 Take 100 Crs of raw propolis, crush it, then reduce it to powder, pass it through the sieve N 3 then N 2.

 Macerate for 24 hours in an opaque bottle with a quantity of alcohol at 96 ° C triple the test portion. Filter.

 Take the solid part remaining on the last sieve, wrap it in a sterile cloth and introduce it into a distillation bottle, topped with a refrigerant.

 Boil with alcohol at 96 C for 1 hour. The quantity of alcohol for an intake of 100 grs will be 500 ml of alcohol at 96 C. Filter the extract, then evaporate in a water bath.

 Resumption by hot water using the same technique, then filtration and concentration in a water bath.

 Collect the extract and place in the refrigerator, in an opaque bottle to be protected from light and firm.

Shake 2 to 3 times a day during storage. The extract obtained is thermostable, it can be autoclaved at 120 ° C for 30 minutes, its pH is close to 7, it has a slight aromatic odor
Propolis extract melts at 80 ° C.

General formula
Purified-concentrated propolis
Ultra-pure water.

 Purified and concentrated propolis alone should be stored before formulation for safe indication, in opaque vials, protected from light and stored out of the refrigerator.

 Conservation for several months (10 to 12 months).

Aqueous extract of propolis
With 100 Grs of test sample as previously indicated in the propolis extraction technique, the following correspondence will be obtained by the concentration of the bain-marie and after filtration: for 1 ml of concentrated extract 62 mg of solids.

 By refractometry it is also possible to determine the quantity of dry matter.

 It is necessary to add in this descriptive technique the propolis extract, to explain its very great interest by the contribution of micro elements, that no physiological or biological process takes place in the human body without the participation. trace elements, that these trace elements participate in a specific way to protein metabolism, carbohydrate, and lipid, protein synthesis, thermal metabolism.

 These trace elements or micro elements are received via food and water, here by the propolis extract in cases where the ground is aggression, or is in imbalance. For example, 24 microelements exist in human blood, iron is used in the composition of respiratory enzymes, zinc in enzymes that participate in protein metabolism, carbohydrate, copper in bone marrow and bone, iodine in the thyroid gland, the nickel in the pancreas etc ...

 In a nutshell the activity of trace elements, biological activity, is characterized by the action in synergy with vitamins and enzymes.

 It has been established and recognized that some propolis have greater effects than others, an average is to be established.

 My in-vivo and in-vitro tests were carried out on this average, by harvesting propolis from different sectors, different regions, so I could notice the concentration stability of the final extract of propolis, by operating on harvests of 50 hives of high populations (more than 60,000 bees) in perfect sanitary condition, taking into account climatic factors, geographical situation, local flora.

 All my tests and results were performed only for the following disciplines or indications; dermatology - cosmetics - antioxidant inhibitory action - preservation - deodorization. I was only partially interested in the certain qualities of the propolis on the medical level not being a doctor, nor specialized in diseases, I did not continue on the other hand if I describe as "certain" the qualities of propolis in medical field, this is in relation to its use in many works carried out by the USSR and Romania. If the medical term "dermatotogy" is used in the indications, it is only valid for the aggressions in certain forms of the skin and that on simple advice or not, you can obtain freely in pharmacy.

 However, the products developed as a result of my trials, have a particular advantage over those existing on the market, namely more effective, without secondary risks, or allergy if one complies with the terms and doses indicated. In addition this extract is a natural substance devoid of toxicity, precise information on its tolerance per kilo of weight have been given in the technical information.

 Anti-bacterial, bacteriostatic properties.

The tests were carried out according to the following experimental technique
For each strain
4 batches of 5 petri dishes (large diameter) in which a culture medium for demanding bacteria (typtone, casein, soybean) was poured before solidification was introduced at 40 ° C progressive doses of propolis concentrate obtained by extraction .

The tryptone culture medium, casein, soy, was carried out at the recommended dose (Institut Pasteur) these progressive documents are
1 lot of 5 boxes at 30 mmg
1 set of 5 boxes at 60 mmg
1 set of 5 boxes at 90 mmg
1 set of 5 boxes of 120 mmg
and this technique for each bacterial strain tested.

Strains are represented by
Subtle Bacillus - Bacillus alvei - Staphylococcus aureus - Streptococcus haemolitricus - Proteus vulgaris. Incubation, t were respected with respect to the characteristics of each type of bacteria.

The results of the tests, researches, showed that all the propolis samples had a pronounced microbial action, equal or superior to certain antibiotics, such as penicillin, tetracycline, streptomycin, and resulting in the following way: for example the concentrate having gave the best result.

<Tb>

<SEP> Propolis <SEP> to <SEP> tetracycline <SEP> penicillin <SEP> streptomycin
<tb><SEP> concentration <SEP> of
<tb><SEP> 60 <SEP> mg
<tb> Staphylococcus
<tb> aureus
<tb> Proteus
<tb><SEP> + <SEP> + <SEP> + <SEP><SEP> ++
<tb> vulgaris
<tb> streptococci
<tb> haemoliticus <SEP> ++ <SEP> ++ <SEP> + <SEP>
<tb> Bacillus
<tb><SEP> ++ <SEP> ++ <SEP> + <SEP> ++
<tb> subtilis
<tb> (+) = not very sensitive (++) = sensitive (+++) = very sensitive () = very effective
The same technique was carried out with the concentrations at 300 - 600 900 - 1.200 mmg, with double culture media in amount of weight.

 The results were identical, which means that propolis even more concentrated in% has no more clear action.

 This test could be more elaborate and studied with other antibiotics and other strains, this is not the goal.

 It was only necessary to demonstrate the bactericidal power of propolis extract on certain strains of microorganisms with standard antibiotics. This bactericidal and bacteriostatic action will not be put in formula to create a product, that is the reason why this study was not carried out more deeply but to demonstrate that the purified propolis concentrate had a bactericidal action and bacteriostatic at very low concentrations.

 Anti-inflammatory, healing action by stimulation of tissue regeneration in dermatology.

 By the presence of micro-elements contained in the concentrate, it will make the process of epithelial proliferation and granulation increase more active, will also improve blood and lymphatic circulation, and reduce the permeability of vessels of the area reached .

 Thanks to its powerful anesthetic properties, its bactericidal effect on gram + and - microbes, its stimulating influence in the regeneration process, propolis extract is particularly indicated in superficial burns - keratodermia of the soles of the feet and hands - boils , external abscesses - wounds with slow or difficult scarring - in bruises, cuts - wounds - warts, frostbite - crevices - horns in feet - scares in general - cutaneous manifestations of shingles.

 The epidermis with propolis extract will get rid in some indications, crusts, the rash will disappear and a thin scar will form. The necrotic mass will be cleaned and the infiltrations will be absorbed very quickly. The granulation tissue will appear, and the opithelium will begin to form. In other specific cases the epidermis will soften and become more elastic, an interesting healing action by stimulation of tissue regeneration will ultimately result in a reduction in treatment time in comparison with the known therapeutic means.

In cosmetics
- In sunburns which are burns in the first degree.

 - in the fine healing sought after surgical procedures, same aesthetic, scars cheloids.

- total or partial hair loss
- diffuse redness of the faces, unsightly pimples.

- seborrheic skin
- fine lines, wrinkles
- intolerances to cosmetic products.

- anti-wrinkle mask
- mask to restore the suppleness of the facial skin and restore freshness, rebalance dilated pores, and tired face.

 Antioxidant quality of propolis extract and preservation.

 Great interest in the food industry, cosmetics, the concentration of total flavonoids present in the propolis extract, as well as terpenescontained in the fraction of essential oils also present.

 Tests were carried out on pork fat which was distilled twice under vacuum, the peroxide content was distilled twice in vacuo, the peroxide content was distilled twice under vacuum, the content of peroxides was was determined by iodometry.

 The following remark was made with the extract containing the sum of total flavonols: the extract significantly retarded the oxidation of the melted pork fat in a ratio of 3 times with respect to the peroxide content. The extract was at a concentration of 10 grs. The extract is therefore an effective natural preservative.

 Inhibitory power of propolis extract or phytoinhibiting power.

 A 2 7 agar-based culture substrate is used with distilled water and propolis extract, which is cast into a 20 cm diameter glass petri dish.

Tests performed in 1982 (April)
The concentrate is 1 part of extract per 100 parts of water, ie 1 g per 100 g of water and the test is carried out on 10 petri dishes for each type of seed.

 In each box we sow 50 seeds of a seed (the tests were made with seed of thought, wallflower, alfalfa, oats, Batavia lettuce) 1 box of kneaded witness to each batch of Seeding does not contain propolis extract.

 The seedlings are distributed in a homogeneous way.

 The oven growth at constant T of 25 C with a percentage in the humidity chamber at 80%. Seedlings are then exposed to light for 30 minutes every 15 hours.

This exposure is repeated 6 times. The light source comes from fluorescent tubes of 40 wats each, the number of tubes will be 10 tubes with spacings between each tube in parallel of 20 in the illumination received at each box must be 1,500
The readings are made after the 6th day. The seedlings are removed (25 per box and the state of the seed is noted).

 The same experiment is done with seedlings from different seeds, they have been grown in agar without inhibitor, 25 seedlings are taken per box, the seedlings are sprayed with the inhibitor (propolis extract) at the stage of the seedling.

 The results for the seedlings will be expressed in lengths of the hypocotyls and lengths of the radicles under a binocular microscope and according to the ratio of the dilution of the extract in the substrate, and the control without extract.

I found
That the seeds had not germinated, that the seedlings at the end of the experiment and after this spraying had had no growth, that it was stopped, without cell destruction. The concentration of the inhibiting power is in the ratio of a dilution 1 part of propolis extract per 100 of water in the case of seedlings. This extract is therefore applicable as a herbicide without any secondary risk at many levels. Herbicides placed on the market do not have this quality.

 Clear action on germination when it is not necessary with a conservation power (for example: seed of potatoes).

 Extract used as deodorant.

- For altered meat with putrid odor
- smell of the mouth (foul breath)
- deodorant (meeting rooms - cinema - premises)
- household garbage (garbage cans, toilets)
The results obtained by the propolis extract are well defined as having a bactericidal, bacteriostatic, inhibitory, anti-oxidant, cicatrizant, anti-inflammatory deodorant and preservative power.

 This research and experimentation spread over 5 years until 1984.

 As regards the tests of using a propolis extract as a preservative with the added quality of being an antioxidant, I can not provide the name of the agri-food company in which the experiments have been made, however I give the technical modality of experimentation namely: at the request of the company interested my commitment not to use this experimentation as a reference by quoting the name of the Company, as long as this product will not have applied for a patent, and other tests on other commodities, for commercial reasons.

The experiment was conducted as follows 3 tests were performed on each batch.

A batch of 20 turkey cutlets (ready to eat),
A batch of 20 turkey roasts (ready to eat),
A batch of 20 broilers (ready to eat),
A batch of 20 meat quails (ready to eat),
A batch of 20 fillets (ready to consume),
A batch of 20 slices of salmon (ready to eat),
These batches before the use of the extract have not been modified, they followed the normal chain of production namely: From the arrival of the pickup truck to the stage of the product ready to be consumed.

This production agro-food technique can not despite all the precautions taken to prevent the presence of bacteria in the premises and other agents, which therefore limits the shelf life of the product wrapped film and ready to be consumed thereafter. The extract will be able to play an important role by its own qualities.

The batches were all sprayed with the concentrate in 2 passes at a spacing of 5 mm between each pass. The lots were 20 units each. These lots were sprayed on a normal table at the production plant and then stored in a room. Spraying for poultry was done externally and internally.

The room temperature was 15'C, it was not maintained at this T ', leaving the free room has its own variations. The duration of the test was 30 days. A control was put under the same conditions without spraying.

2nd test). The batches were sprayed on a table as before but with a single pass, for poultry a single spray also internal and external, the temperature of the premises at departure was 17 ', but this one as for the previous test to give an extra value to the quality of Propolis extract was not retained, leaving the free room to variations. The lots were 20 units each, the duration of the test was 30 days. A control was put in the same conditions without spraying.

3rd trial). Lots of 20 units each, single pass spray for external and internal poultry. A control was put under the same conditions without spraying for 15 days. These batches were put in cold room and then out of it and placed in 4 ovens at a temperature of 40'C (4 ovens by a significant number of batches) maintained for 15 days at this constant temperature.

The following results were obtained
No alteration of the meat after cutting. No particular smell, even sensitive, at the tasting no change of taste or smell. In addition it was found that only one spray was needed.

This experiment demonstrates the additional conservation possible with this propolis extract in foodstuffs. It is obvious that other tests are needed and on other commodities, I will also test this extract of propolis cheese, to give some pie charts more resistance to casting, also on yogurts, small Swiss, which ' have very limited conservation limits.

I request the protection of my applications by the propolis extract for the following industrial activities
Cosmetics Industry, Food Industry, Plant Protection Products Industry, Dermatology and Hygiene and Deodorization Products.

According to the following formulas
Indus sorts cosmetics
Indication: partial or total hair loss. Formula for 100 gr 1 gr extract - 99 gr ultra pure water PH neutral.

Instructions for use: daily frictions on the head at night, keep at night, schampoing plants (neutral) every morning.

Presentation: aerosol form, with nitrogen propellant or 125 ml glass opaque bottle.

This neutral PH product can be used for any type of hair.

It can not create a secondary risk or allergy. It is a stimulant of first quality, it stops more hair loss.

Indication scars cheloides fine scarring following surgical or aesthetic surgery, wrinkles and fine lines, sunburn (curative and preventive).

Cheloid scars: extract 2 grs, ultra pure water 98 grs, PH neutral.

wrinkles fine scars following surgical procedures. Fine lines
Sunburn
Formula for 100 gr.

0.500 c of propolis extract - 99.5 grs ultra pure water
Neutral PH
How to use: Spray scars (twice a day) in the morning and evening.

Wrinkles and fine lines ditto.

Presentation: aerosol with nitrogen propellant
masks
Technique of manufacture: Dissolve in the ultra pure water and in a sterile way the following part: Boric acid, then add the Bentonite when the temperature of the water during the cooling will have reached 50'C. Stir for 15 minutes, then add the concentrate when the mixture reaches 30C.

Formula for 100 grs
Boric acid 0.50 grs
Bentonite 5 grs
Concentrated 1 gr
Ultra pure water 93 grs 5
Neutral PH
Bentonite is used in this formula for its absorbency, its pore cleansing activity and impurities of the skin and releasing to restore the skin a clear complexion, radiance, softness.

Indications: Facial skin, pronounced attenuation of wrinkles and fine lines, restores radiance and softness, deep cleansing of the epidermis, excess of sebum resulting in shiny and oily skin, bringing a significant freshness of the face.

How to use: This mask is to be used according to his desire and need for treatment every day for 8 days and leave in application 1 hour on the face.

For pronounced wrinkles and fine lines, the treatment should be increased to 1 month.

It is recommended to remove the mask with neutral cleansing milk, or distilled water, do not use tap water.

Presentation: In opaque glass pot.

This mask by its composition and originality is a product of high quality far superior to those existing on the market. It can be used for any type of skin, without any secondary risk.

Phytosanitary products industries
Phytoinhibitor potency of propolis extract
As a herbicide
Formula for 100 grs, 10 grs of extract for 90 grs of ultra pure water.

Then lyophilization for its spray application, tap water, well water can be used.

Indication
The propolis extract by its own qualities will stop the growth of weeds so particularly indicated in cases where this action is necessary. It is a new and major weed control agent with the following advantages over all other existing products, can not cause any pollution or harmful infiltration at groundwater level, does not create for animals, bees, and restriction manipulator particular, of secondary risks. In addition, this product is devoid of any toxicity, it is natural, PH neutral.

Manual
By spraying
Presentation in freeze-dried powder
Germination inhibitory agent
Same condition for use and benefits, the presentation as the herbicide product.

Formula for 1 000 ml, 2 grs of propolis extract, 998 grs of ultra pure water, followed by freeze-drying.

Agro Food Industries
Formula: 10 grs solution of propolis extract for 990 ml of ultra pure water and freeze-drying.

How to use: spray
Indications: Conservation of solid foodstuffs. For products such as yogurts, Swiss swiss, cheese, the tests are not completed, but already giving positive results, freeze-dried propolis extract must be introduced before caking or even before 50'C and put with stirring.

Deodorant: Hygiene
indications
Altered meat with putrid odor
Bad household odors (WC, various premises, garbage cans)
Meeting rooms, cinemas, industrial premises, offices.

Smell of the mouth (foul breath and refreshment)
How to use: In spraying.

Presentation: Freeze-dried product
For the smell of the mouth the concentrate must be presented in small pressurized bottle (model of pocket or purse) with propellant with nitrogen.

Indications: Impaired meat with putrid odor, bad house smells.

How to use: spray.

Formula for 1000 ml, 20 grs propolis extract, 980 grs ultra pure water.

Lyophilization then.

indications
Meeting rooms, cinemas, offices, industrial premises
Formula for 1000 ml, 5 grs of propolis extract, 995 grs ultra pure water.

Use: spray.

Presentation: aerosol or freeze-dried powder.

indications
Smell of the mouth
Formula for 1000 ml, 0.5 grs of propolis extract for 995.5 ultra pure water.

Aerosol presentation, nitrogen propellant.

Dermatology
indications
Keratodermia of the soles of the feet and hands.

Bedsores in general, warts, corns in feet, frostbite, bruises.

Formula for 100 grs, propolis extract 10 g.

Vaseline pure codex 80 grs, Lanolin anhydrous 10 grs.

Manual
Apply the ointment in the morning and evening, wrap with a cloth or gauze impregnated with the product.

Presentation :
In pot type opaque glass ointment jar.

Indications can also be aerosolized.

Aerosol formula for 1000 grs
Propolis extract 10 grso ultra pure water 990 grs.

Presentation: nitrogen gas propellant aerosol Indications
Superficial burns, boils, external abscesses, wounds whose healing is slow, difficult. Cutaneous manifestations of shingles, cuts, wounds, crevices.

Manual
In application on the part to be treated twice a day, do not wrap.

Presentation
In pot type ointment jar (opaque glass).

Formula for 100 grs (presentation ointment) 3 grs propolis extract
Vaseline pure codex 85 grs
Lanolin anhydrous 12 grs
Formula for 100 grs (aerosol presentation)
Propolis extract 3 grs
Ultra pure water 97 grs
Nitrogen propellant
How to use: spray.

Methods of manufacturing formulas used in dermatology
Melt Lanolin and Vaseline separately in a bain-marie, allow to cool, add the propolis extract to 45 C, shake to distribute, mix and let cool until completely solidified.

NOTE: These formulas have been studied, and have been tested in adults only.

Means of control :
Determination of micro elements by spectrophotometry and codex method, usual technique.

Thin layer chromatography, usual technique.

Claims (1)

 RENENDICATIONS  - The process of extraction of propolis characterized by maceration for 24 hours in a container or opaque bottle with a quantity of alcohol at 96 C triple the weight of the sample, this maceration is then filtered, the solid part is wrapped in a sterile cloth introduced into a distilling bottle to be overflowed with a condenser, then boiled with alcohol at 960C for 1 hour, the volute of which will be 500 ml per 100 grams of propolis, this preparation is again filtered and evaporated in a water bath, an extract is obtained which is taken up in hot water by applying the technical distillation name as indicated above, and then filtered and concentrated in a water bath.