LIQUID MIXTURES

Technical Field

The invention relates to propolis products.

The invention relates in particular to liquid propolis products which can be used in many sectors such as food, health and cosmetics that offer them for human consumption in their most suitable and in the highest form of bioavailability, in a fast and easy consumable state by properly extracting propolis which cause development of antioxidant, antimicrobial, antifungal, antivirus, anti - inflammatory, anti - cancerogen and anesthetic effects as well as many beneficial biological activities, and production methods thereof.

Prior Art

Propolis is a bee product which the bees process the resinous substances that they collect from the buds and bodies of plants. The bees produce propolis for protecting the hive against micro - organisms and external factors by repairing the cracks in the hive with propolis. The bees use the resins of mostly poplar and pinaceae while producing this. There are 45 % resin, 30 % beeswax, phenolic compounds, 5 % minerals and vitamins in the composition of propolis. Propolis basically has antimicrobial, anti - inflammatory, antioxidant, anti - cancerogen and anesthetic effects, and it provides supporting the immunity system as well. Because of the fact that it is being used frequently in important fields such as medical, apitherapy, healthy nutrition and biocosmetic fields, it is a rather popular component. Propolis basically comprises 150 chemical compounds, more than 20 minerals, beeswax, resin and pollen.

Propolis which is produced by bees to protect their hives cannot be consumed as it is taken from the hive because it comprises high content of beeswax and parasites which try to reach the honey in the hive. In order for propolis to be converted to a suitable form for human consumption, it is necessary to remove the beeswax and impurities in its content. Following removal of beeswax and other impurities from propolis with its bioavailability of 2 % in its raw state, the bioavailability can rise up to 100 %. And our body employs propolis that converted to completely pure form composed of antioxidant substances such as phenolic and flavonoid.

Chinese patent application number CN105560289A relates to the preparation technique of liquid propolis. Said technology includes the following steps: disintegration of propolis raw material, preparation of a liquid mixture composed of caprilic / capric triglyceride, propylene glycol, glycerin, pure water and an amount of alcohol mixture, mixing both components, filling liquid propolis mixture and packaging. There is the probability of encountering some side effects in the propolis dissolved in glycerin too. Since glycerin cannot be completely absorbed in the body, it can be harmful to consume it too much alone or together with food. Also, extraction processes carried out with lipid kinds such as triglyceride turn out to be slow and cannot give the desired performance.

Turkish patent application number TR 2019 / 20656 which is another application present in the literature relates to water based liquid propolis in which in its content, antioxidant value has been increased by the addition of boron that has been prepared in a form suitable for human nutrition and its liquefaction has been provided by a special technology. This composition too can cause various side effects from the point of view of health because of the boron compounds.

Many products comprising propolis are being sold in the market. As a result of analyses made on these products, it has been seen that propolis content information written on the label is wrong, or extraction methods are insufficient. It is necessary for the raw propolis to be extracted correctly due to the indigestible components such as impurities and beeswax in its content, and to be supplied to consumption in its highest state of bioavailability by preserving the beneficial components in its content in the maximum amount. Antioxidant content and bioavailability of the products in the market are rather low. In addition, the composed mixtures comprise components which can be harmful to human health. The insufficient content of these products which are bought to increase body resistance and to fight with diseases is not suitable for the purpose.

Moreover, direct consumption of propolis is rather difficult due to its bitter taste and resinous structure. Therefore, products available in the market use many additives and chemicals to overcome this consumption difficulty. The purpose of using these chemicals is to bring in solubility to the resinous structure and to try to mask the bitter taste with the artificial sweeteners used. Moreover, the firms are unable to purify propolis correctly or import it from countries such as China. Residue and additive problems are frequently encountered in the imported propolis. Consequently, the firms are obliged to add chemical preservatives and additives to the final product due to the fact that they cannot make use of the antioxidant, antimicrobial, antifungal and similar bioactive effects of propolis. And this in turn causes the product to attain an undesirable condition.

Consequently, due to the above mentioned negativities and because of the insufficiency of the existing solutions about the subject it has been necessary to perform an improvement in the related technical field.

Brief Description of the Invention

The present invention relates to liquid propolis products which meet the above mentioned requirements, which eliminate all disadvantages and which offer some additional advantages, and its method of production and liquid propolis product.

The prioritized purpose of the invention is to provide the method to obtain a properly extracted liquid propolis extract with high bioavailability and liquid propolis product.

An object of the invention is to provide a liquid propolis product and a method of obtaining liquid propolis extract without side effects or drug-food interaction.

Another purpose of invention is to form the steps applied during the method in a way not to disturb the stability of propolis product in a suitable and efficient result manner.

Another purpose of invention is to obtain properly extracted liquid propolis extract with high flavonoid content which causes development of antioxidant, antimicrobial, antifungal, antivirus, anti - inflammatory, anti - cancerogen and anesthetic effects as well as many beneficial biological activities.

Another purpose of invention is to provide raw materials or final products from the obtained liquid propolis extract to be used for healthy nutrition, apitherapy and biocosmetics purposes in the food, health and cosmetics sectors. In order to accomplish the above mentioned objects, the invention relates to a method of obtaining liquid propolis product which comprises the following process steps and which has high antioxidant capacity (a) disintegrating the raw propolis by passing it through a grinder; (b) mixing the disintegrated propolis with at least one fruit juice selected from a group consisting of ethyl alcohol, propylene glycol, ultra pure water, balsam apple and black elderberry extract and a solvent selected from a group consisting of carob extract; (c) mixing the obtained solution by adding lemon juice or apple cider vinegar as acid regulator; (d) shaking the degassed solution; (e) degassing the shaken solution in ultrasonic bath; (f) centrifuging the degassed solution; (g) removal of volatile substances by drying the centrifuged mixture in a rotary evaporator until the extent of dryness; (h) carrying out analysis; and comprises the process steps of

(i) Addition of honey, ginseng and / or royal jelly and at least one fruit juice selected from the group consisting of balsam apple, black elderberry extract to the mixture in which solvents have been removed by drying and mixing the same;

(j) Addition of perga, pollen, salt, menthol and / or spices to the mixture;

(k) Mixing the mixture in a homogenizer;

(L) Degassing it in an ultrasonic bath. The invention also includes the propolis products obtained by the above mentioned method and the use of these products for healthy nutrition, apitherapy and biocosmetic purposes in food, health and cosmetics sectors.

Structural and characteristic features of the invention and all of its advantages will be understood more clearly by means of the detailed description given below and therefore it should be appreciated by considering this detailed description.

Detailed Description of the Invention

In this detailed description, liquid propolis products of the invention and production methods thereof are described aimed at only better understanding of the subject and without forming any limiting effects. The invention relates in particular to liquid propolis products which can be used in many sectors such as food, health and cosmetics that offer them for human consumption in their most suitable and in the highest form of bioavailability, in a fast and easy consumable state by properly extracting propolis which cause development of antioxidant, antimicrobial, antifungal, antivirus, anti - inflammatory, anti-cancerogen and anesthetic effects as well as many beneficial biological activities, and production methods thereof.

The invention is also a liquid propolis extract suitable for healthy nutrition, apitherapy and biocosmetic purposes for use in food, health and cosmetic sectors offered as drops, syrup or drinks wherein liquid propolis obtained by extracting pure propolis with of at least one fruit juice selected from ethyl alcohol, propylene glycol, ultrapure water, potion pomegranate and black elderberry extract; and a solvent selected from group consisting of carob extract; and lemon juice or apple cider vinegar as acid regulator comprises honey, ginseng, royal jelly, pomegranate, at least one fruit juice selected from black elderberry extract, perga, pollen, salt, menthol and/or spices.

Since the raw propolis cannot be digested directly due to the indigestible components such as impurities and beeswax in its content; it has to be freed of the impurities by extracting with a solvent. In Table 1 , preferred raw materials of the invention and their amounts in the method of invention are given. The liquid propolis product which has been developed accordingly comprise honey preferably in an amount from 12.5 % to 37.5 % (more preferably 25 %) based on total weight, salt preferably in an amount from 12.5 % to 37.5 % (more preferably 25 %) based on total weight, ginseng preferably in an amount from 0.5 % to 2.5 % (more preferably 1 %) based on total weight, royal jelly preferably in an amount from 0.5 % to 2.5 % (more preferably 1 %) based on total weight menthol preferably in an amount from 0.05 % to 0.5 % (more preferably 0.25 %) based on total weight, spices preferably in an amount from 0.05 % to 0.5 % (more preferably 0.25 %) based on total weight, pollen preferably in an amount from 0.5 % to 2.5 % (more preferably 1 %) based on total weight, - perga preferably in an amount from 0.5 % to 2.5 % (more preferably 1 %) based on total weight.

Table 1. Raw materials used in the method of the invention

Raw propolis: Propolis is a plantal substance in the form of a sticky resin which is collected by bees from the trees and carried to the hives. It is a bee product having natural antibiotic property. Propolis which has a natural antibiotic property causes implementation of antioxidant, antimicrobial, antifungal, antivirus, anti - inflammatory, anti - cancerogen and anesthetic effects as well as many useful biological activities. It reinforces the immunological system. It is known to be effective against cancer, inflammatory diseases, diseases of viral and bacterial origin. The raw propolis should be freed of the impurities by extraction since it cannot be directly consumed due to the indigestible components such as impurities and beeswax in its content. Therefore, the raw propolis should be extracted with the correct solvents.

Solvent: Raw propolis should be freed of the impurities by extraction since it cannot be directly consumed due to the indigestible components such as impurities and beeswax in its content. Solvent is the agent which provides the dissolution of the raw propolis and transport of the active materials. In the method of obtaining the propolis extract of the present invention, as a solvent at least one fruit juice selected from the group consisting of suitable for use in foods ethanol (ethanol) with a purity of more than 90%, propylene glycol, pomegranate and black elderberry extract; carob extract or ultrapure are used. Fruit juices mentioned herein can use different solvents which are not harmful against human health according to the form and composition of the desired final product to be obtained by pomegranate juice, orange juice, strawberry juice, apple juice, peach juice, sour cherry juice or similar fruit juices. Solvents which are not suitable for human health such as methanol, ethyl acetate, chloroform and hexane are not preferred in the invention.

Ethanol, ethyl alcohol or plantal alcohol is a colorless chemical compound obtained from plants. It is currently being used as solvent in vitamin syrups.

Ultra pure water is the more purified state of pure water. Pure water is the water in which mineral substances present in the water have been removed. And ultra pure water is water with a conductivity value of 0.055 pS / cm (25 Ό) or electrical resistivity of 18.2 Megaohm - cm.

Propylene glycol is a petroleum oil derivative compound that has many uses in the commercial industry. It is odorless and it does not have any certain taste definition. Propylene glycol is being used in food, cosmetics, pharmaceutical and plastics industries. Ethyl alcohol and propylene glycol are suitable to be used in foods and are not subjected to any dissolver heating process.

Balsam apple extract Balsam apple (Momordica charantia) is being used in diabetes patients as an alternative treatment for lowering the glycose levels in the blood. Research on animals and humans show balsam apple to comprise hypoglisemic substances. In a study made on mice, the results have shown a significant increase in b cell count in the animals treated with M. charantia as compared to untreated diabetes patients. M. charantia extracts are being used as a potentially cytotoxic agent for certain cancer types in addition to being used in clinical conditions such as diabetes, dyslipidemia, microbial infections. In a study to assess the efficacy of balsam apple extract, human breast cancer cells, MCF - 7 and MDA - MB - 231 and primary human breast epithelial cells have been used as an in vitro model. The treatment of breast cancer cells with balsam apple extract has resulted in a significant decrease in cell proliferation and induced apoptotic cell death. This study shows that balsam apple extract modulates the signal pathways for the inhibition of breast cancer cell growth and that it could be used as a diet supplement for preventing breast cancer.

Black elderberry extract is a good source of protein, and they are the free and conjugated form of amino acids, unsaturated fatty acids, fiber fractions, vitamins, antioxidants and minerals. Polyphenols comprise mostly anthocyanins, flavonols, phenolic acids and proanthocyanidins as well as high biological activity components such as terpens and lectins. Anthocyanins and other flavonoids show antioxidant, anticarcinogenic, immuno - stimulating, antibacterial, antiallergic and antiviral properties; therefore their consumption can contribute to the prevention of various degenerative diseases such as cardiovascular disease, cancer, inflammatory diseases and diabetes. Acid regulating substance is being used to increase the acidity of the solution consisting of propolis and solvent in the method of obtaining high flavonoid content propolis extract of the invention. When ethanol, propylene glycol or ultra pure water mixture is being used as solvent, preferably 1 ml of lemon juice can be used as acid regulating substance. When propylene glycol, carob extract or at least one of the above mentioned fruit juices is being used as solvent, preferably 1 ml of apple cider vinegar can be used as acid regulating substance. Different acid regulators too can be preferred for different solvents other than these.

Ginger, turmeric and cinnamon are preferably being used as spices in a preferred embodiment of the invention. There is a great number of published reports related to the antioxidative activity of these spices. Spices are food supplements that have been used as flavoring and coloring agents and as preservatives for thousands of years. The digestion stimulating effect, hypolipidemic effect, antidiabetic effect, antilithogenic property, antioxidant potential, anti - inflammatory property, antimutagenic and anticarcinogenic properties of the spices have been documented. Among these, hypocholesterolemic and antioxidant properties of a few specific spices have a wide range nutraceutical value. These useful physiological effects also have possible therapeutical treatment potential under various disease conditions. Turmeric (Curcuma longa) is used in many urinary disases, liver diseases and in jaundice. Turmeric extract and curcumin isolated from it are rather cytotoxic in mammalian cells and are effective in reducing animal tumors and shows potential of usage in cancer treatment. Ginger (Zingiber officinale Rose.) is one of the richest sources of antioxidants which cleanse the free radicals and the health inconsistencies that they bring along with them. Ginger comprises gingerols as active components which exhibit antioxidant activity determined by in vitro studies.

Pollen is a product of a great veriety and rich in biologically active substances. In pollen grains from different plant kinds 200 substances have been found. Proteins, amino acids, carbohydrates, lipids and fatty acids, phenolic compounds, enzymes and coenzymes, vitamins and bioelements are included in the fundamental chemical substances. Bee pollen is used in apitherapeutical treatment since it shows a series of actions such as antifungal, antimicrobial, antiviral, antiinflammatory, immuno-stimulant and local analgesic action and also because it facilitates the granulation process of burn wound healing. Bee pollen is a precious nutritive substance and a natural product of honey bee advertized as a source of energy. The health promoting value of bee pollen originates from the presence of enzymes and co-enzymes in the bee pollen as well as secondary plant metabolites (tocopherol, niacine, thiamine, biotine and folic acid, polyphenols, carotenoid pigments, phytosterols).

Among the active components found in ginseng include ginsenosids, polysaccharides, peptides, polyacetilenic alcohols and fatty acids. There is a wide variation in the ginsenosid content of different ginseng species. There are many clinical studies on ginseng to characterize its therapeutic properties such as improvement of physical performance, cancer treatment, diabetes and hypertension.

Royal jelly is a very precious bee product having a great number of functional properties such as antibacterial activity, anti - inflammatory activity, vasodilatative and hypotensive activities, disinfectant effect, antioxidant activity, antihypercholesterolemic activity and antitumor activity.

Salt currently plays a multi-purpose role in many produced foods and drinks in addition to its roles in human physiology, nutrition and health. Salt which encompasses sodium and chlorine in its content is an important compound in meeting the sodium requirement of the body in addition to giving flavor. Salt also possesses specific properties which are important for the processing of these products in many food products. Salt possesses an effect primarily on the water activity and a protective duty against microbes generally in combination with other antimicrobial agents in addition the mentioned properties. In addition to its protective duty, it provides stability of the product by effecting the acid base balance. Salt content in the products not only effects their tastes but also their microbiological shelf life and in some instances their texture and mouthfeel too.

Menthol, plant oriented menthol is the product of natural distillation of mint leaves. Menthol which is structurally similar to mint is denser in terms of elements that it comprises. Menthol which is used in food usually to give aroma is also antiseptic for the intestines with its anti - inflammatory and vasodilatator property. It plays an important role in the treatment of respiratory tract diseases. Menthol is being preferred in respiratory tract ailments and in allergical situations in nasal sprays with its anesthetic and deconjestant properties.

Perga is pollen in the form of pellets collected by the bees and carried back to the hive by mixing them with their own digestive enzymes, comprising a little amount of honey and beeswax. By means of fermentation implemented by the enzymes of the bees, perga which is formed by melting of the outer membrane of the pollens encompasses many useful microorganisms thereby showing probiotic property. Perga which comprises many vitamins, phenolics and antioxidants is also being used as a natural food supplement.

The invention comprises honey as a sweetener. Honey is a natural product that changes the plant nectars, secretions in the living parts of the plants or the secretions of the plant sucking insects living on the living parts of the plants by combining them with specific substances after being collected by the honey bee, reducing the water content and maturing by storing with honeycomb. Honey which is a source of vitamins and minerals has a lot of benefits to the body, especially in the immunological system.

The method of obtaining liquid propolis product of the invention comprising the process steps of (a) disintegrating the raw propolis in amounts indicated in Table 1 by passing it through a grinder; (b) mixing the disintegrated propolis with at least one fruit juice selected from the group consisting of balsam apple and black elderberry extract, with a solvent selected from a group consisting of carob extract or ultra pure water preferably in amounts indicated in Table 1 ; (c) mixing the obtained solution by adding lemon juice or apple cider vinegar as acid regulator; (d) shaking the degassed solution; (e) degassing the shaken solution in ultrasonic bath; (f) centrifuging the degassed solution; (g) removal of volatile substances by drying the centrifuged mixture in a rotary evaporator until the extent of dryness; and (h) carrying out analysis and having a high antioxidant capacity in its most fundamental form comprises the following process steps; (i) mixing by adding at least one fruit juice selected from the extract of pomegranate, black elderberry; honey, ginseng and / or royal jelly to the mixture removed by drying the solvents;

(j) Addition of perga, pollen, salt, menthol and / or spices to the mixture;

(k) Mixing the mixture in a homogenizer;

(L) Degassing it in an ultrasonic bath;

In an embodiment of the invention, when the above mentioned ethanol, propylene glycol or ultra pure water is used as solvent, lemon juice can be used as acid regulating substance in the amounts given in Table 1. When propylene glycol, carob extract or at least one of the above mentioned fruit juices is used as solvent, apple cider vinegar can be used as acid regulating substance in the amounts given in Table 1.

A sample is taken from the supernatant which is formed following the centrifuging process, and bioavailable phenolic flavonoid substance content which the propolis comprises is analyzed in LC - MSMS (Liquid Chromatography - Mass / Mass Spectrometer). In the analysis results, the condition of galangine, pinocembrin, crisine, cinnamic acid to be above 10 mg / g and p - coumaric acid, ferulic acid, cafeic acid phenethyl ester (CAPE) to be above 5 mg / g is sought. In the event when the analysis results are above the desired values, dry matter determination of the sample is carried out. In the event when the analysis results are not suitable, said solution is again subjected to the processes of continuous stirring in a medium temperature of 20 - 30 ^< C for at least 7 d ays in darkness; degassing in an ultrasonic bath for 3 hours at 25 ‘C; centrifuging at 10 000 g for 20 - 30 minutes, in sequence. A new sample is taken from the supernatant which forms as a result of centrifuging process and phenolic flavonoid substance content is analyzed again in LC - MSMS.

In the obtained solution mentioned above, if the amount of dry substance has reached the desired ratio, at least one fruit juice selected from the extract of black elderberry extract; honey, ginseng and royal jelly are added to the mixture that solvents are removed by drying. Perga, pollen, salt, menthol and / or spices are added to the mixture. The mixture is stirred in a homogenizer preferably at 30 ‘C preferably for 15 - 30 minutes and is degassed in an ultrasonic bath. Filling and packaging of the final products are carried out to be offered to the health and cosmetics sector for the purpose of healthy nutrition, apytherapy and biocosmetics. Useful components that are present in the propolis are preserved by the method described above, and products which have high bioavailability, which are suitable for human consumption, in different liquid forms such as drops, syrups, drinks and in consumption ways can be produced.

Children and adults can consume the liquid propolis products of the invention since they are natural and are source of health. Said product is being consumed as 150 - 200 ml drinks, and it is proper for adults to consume 2 bottles daily, and for children 1 bottle daily. Also, the mixture comprising propolis, royal jelly and ginseng is suitable to be consumed by the adults as 25 - ml shots. The syrup form of the product is suitable for consumption of children and 10 - ml consumption as 5ml ^* 2 daily is being reccomended. Daily intake dose for all liquid products of the invention can vary depending on the propolis amount in its content.