PRODUCT MADE BY THIS METHOD

Technical Field

[0001] The invention concerns the method of producing a functional product of berries made into a puree with crushed seeds and skins and the device for producing the product. The invention also applies to the functional product of berries made into a puree with crushed seeds and skins made in this way and enriched with the content of berry seeds and skins, namely, with polyunsaturated fatty acids, fat-soluble vitamins and flavonoids, which remain unavailable under conventional processing conditions.

[0002] The berries mentioned belong to the genus Vaccinium and include red bilberries, high-bush whortleberries, common whortleberries, cranberries, lingonberries (Vaccinium praestans), blueberries.

[0003] Other berries such as strawberries, raspberries, black currants, red currants, sea- buckthorn, blackberries, etc with seeds up to 2.5-3 mm can also be processed by this method. The same holds true for other fruit and berries, including tropical ones.

Background Art

[0004] Physical and chemical characteristics as well as biological activity of flavonoids, especially polyphenols and anthocyanins, present in the berry skins and seeds have been thoroughly studied for years.

[0005] Anthocyanins make a special class of natural flavonoids which impart red, purple and blue color to various fruit. Of late, anthocyanins have become especially important as dietary antioxidants. For example, blueberry (Vaccinium myrtillus) anthocyanins have been widely used for improving visual acuity and for treating disturbed circulation. Experimental data have been obtained on flavonoids anti-inflammatory properties. Besides, there is information confirming that orally administered anthocyanins are beneficial in treating diabetes and ulcers, in addition they are known for their antiviral and antimicrobic action.

[0006] Pro-anthocyanidins make another class of natural flavonoids which are contained in berry seeds and skins. Recently medical research has been performed on therapeutic application of pro-anthocyanidins which are famous for their antioxidant activity. Besides, there has been information on these compounds antiviral, antibacterial, anticancerogenic, antiinflammatory, antiallergic and vasorelaxant action. They have been shown to prevent lipid peroxidation, platelet aggregation, capillary penetration and fragility and to influence fermentation system. For example, monomeric pro-anthocyanidins (i.e. anthocyanins) and bi- variate pro-anthocyanidins are used for treating diseases caused by capillary fragility, besides, they are known for their anti-inflammatory effect with animals (Beladi, et al., Ann. N.Y. Acad. Sci., 284; 358 (1977)). The published data testifies to the fact that oligomeric pro- anthocyanidins are useful in treating various abnomalities (Altem. Med. Rev. 5(2): 144-151 (2000))

[0007] Pro-anthocyanidins can also ensure antiviral protection. Laboratory experiments have shown pro-anthocyanidins of Virginia hazel (Hamamelis virginiana) to neutralize herpes virus (HSV-1). (Erdelmeier, CA, Cinatl, J., Plant Med. June: 62 (3):241-5 (1996) DeBruyne, T., Pieters, L.J. Nat. Prod. July: 62 (7):954-8 (1999)).

[0008] Because of the above characteristics and anthocyanins and pro-anthocyanidins usefulness, there have been made numerous attempts to extract these compounds from fruit, vegetables and other plants. But apart from anthocyanins and pro-anthoacyanidins, plants, fruit and vegetables contain other compounds such as mineral salts, common organic acids, for example, citric and tartaric acids, carbohydrates, glycosides, catechines.

[0009] At present flavonoids are extracted from plants and fruit by various methods. The methods of flavonoids extraction and purification include heating the aqueous plant material, its ultrafiltration, this being performed at very high temperature, which can ruin flavonoids. Besides, ultrafiltration causes low-molecular polyphenol material separation from the end product.

[0010] Several methods of anthocyanins and pro-anthocyanidins extraction and separation include the use of toxic and environmentally hazardous materials. Thus, the known methods of flavonoids extraction and purification can hardly be adapted to the industrial production scale where removal of various chemicals and solutions is very important. Besides, anthocyanins and pro-anthocyanidins are to be extracted in such a way as to minimize their natural tendency to destruction.

[001 1] Flavonoid extracts are used as active ingredients of food additives in the form of tablets or capsules. But the schedule of tablets and capsules intake is to be strictly observed, which sometimes presents difficulties for people.

[0012] It is better when people take flavonoids in with food, but it is still better when flavonoids are available in their natural form without extraction and further introduction into food. Such product becomes feasible after crushing plant seeds and skins (for instance, those of berries), thus making their useful content available. [0013] The seed contents of such berries as blueberries, bilberries, cranberries, etc (Vaccinium class) are very similar and contain very important ingredients, such as

polyunsaturated fatty acids (PUFA), especially a-linolenic PUFA (18:3) which is better known today as Omega-3. The efficiency of PUFA digestion rises substantially in the presence of fat- soluble vitamin E and β-carotene, they being also part of berry seeds.

[0014] At present marketed Omega-3 is extracted from cod-liver oil but its world reserves have been shrinking considerably of late. The technology of Omega-3 extraction and packing does not prevent its partial oxidation, which is conducive to its fast spoilage and loss of its valuable properties. Besides, the process of oxidation brings about the loss of valuable fat- soluble vitamins.

[0015] The technologies of blueberry natural juice production are well known (see http://www.vandykblueberries.ca/) but they consist just in squeezing out the juice and removing skins and seeds. Under industrial conditions blueberry pulp juice (dispersion juice) is conventionally produced by pulping machines of various design and employing various size sieves. Previously heated flattened blueberries are fed to the pulping machine where they are pushed through a sieve by centrifugal force leaving their seeds on the sieve. This causes 22- 25% loss of valuable feedstock, mainly in the form of berry skins.

[0016J Method and device for hydrodynamic processing of juicy fruit is described in international application WO/2004/056213. This method consists in circulating berries along a closed circuit: pump - turbulence (cavitation) device - flow apparatus - pump until homogenous mass with 100-300 μιη particles is obtained.

[0017] Method and device for hydrodynamic processing of plant seeds is also described in USA patent No. 7,428,797 B2. The method described in the patent consists in circulating soybeans and water along a closed circuit: pump - turbulence (cavitation) device - flow apparatus - pump until homogenous mass is obtained. The device performs loading an axially symmetrical vertical flow apparatus with soybean slurry in an aqueous medium until the circulation circuit is filled, which requires a pump and piping; soybean dispersion by pumping slurry within closed circuit ensuring flow turbulence before its entering the flow apparatus and slurry spiral twisting inside the apparatus until the required consistency and temperature are obtained; degassing and discharge of the product. To avoid protein and fat destruction and oxidation, the slurry is twisted to form a spiral, its radius decreasing from top to bottom, and then directed to recirculation through an opening in the lower part of the flow apparatus;

degassing is started only after an axially symmetrical cone-shaped cavity in the slurry is formed; on achieving the required homogeneity and temperature of the entire product volume the circulation is stopped and the circulation circuit is emptied.

[0018] An attempt has been made to apply the device to continuous production of puree from juicy berries with seeds. For this flattened fresh berries are continuously fed to the device by an additional pump mainly into the sucker of the principal pump. As a result of repeatedly circulating flattened berries and their juice through the turbulence or cavitation device, the berry skins are ruined and heated to the pureeurization temperature.

[0019] The slurry flow twisting in the flow apparatus brings about the removal of air contained in berries through the cone-shaped cavity inside the rotating slurry. The removal of the air is performed continuously and simultaneously with heating and crushing at about 80°C.

[0020] But as the experience has shown, this technology and device do not ensure berry skin homogenous crushing, let alone berry seeds, which results in poorly crushed or even uncrushed seeds getting into the finished product. This is due to the fact that the berry puree viscosity is dozens of times higher than that of freshly crushed ones. That is why the flow fractioning device in the form of the flow apparatus tangentially twisting the flow is not very efficient for making berry puree with crushed seeds and skins, as it does not ensure complete crushing of berries or their parts that can pass through the device without circulation and do not get into the turbulence or cavitation zone a sufficient number of times.

Disclosure of the Invention

[0021] Proceeding from this it is necessary to invent a method of extracting compounds (especially from seeds and skins) containing phenol compounds such as pro-anthocyanidins as well as PUFA together with fat-soluble vitamins to be used in nutraceuticals. This method is to be economically effective for industrial production, economically justified, denying the use of toxic solutions or agents and enabling the extraction of the above compounds in such a way as to decrease their natural tendency to destruction. Thus, there is a demand for a device ensuring such process and the product resulting from the process.

[0022] This demand is satisfied by the method for producing a functional product from berries in the form of puree with crushed seeds and skins which consists in feeding the raw stuff made by flattening berries to obtain pulp, that is, juice, seeds and flattened skins, into a closed hydrodynamic circuit at its initial temperature where simultaneously with air removal the stuff is circulated under pressure pulsations, turbulent friction and cavitation bulbs collapse which causes heating; this destroys and crushes skins forming suspension with uncrushed and/or coarsely crushed seeds; according to the invention the stuff circulation continues until the suspension temperature rises to the previously set value depending on a specific type of stuff characteristic of suspension with particle size less than the characteristic size of the seeds; after that under the centrifugal force the flow twisting separates the stuff with uncrushed seeds from the stuff with crushed seeds and directs the stuff with uncrushed seeds to a bypass channel which connects the centrifugal separator and the pump sucker, thus bypassing the flow apparatus; this makes an additional circuit which includes a bypass channel, a pump, a hydrodynamic baffle, a centrifugal separator and a bypass channel, this additional circuit length being not longer than the closed hydrodynamic circuit length; the circulation of the suspension portion which enters the bypass channel continues in the additional channel, the circulating factor being higher than the suspension circulating factor in the closed hydrodynamic circuit; the circulation in the closed and additional circuits continues until the suspension temperature in the closed hydrodynamic circuit rises to the sterilization temperature, the air contained in berries is removed from the suspension which has achieved the starting sterilization

temperature; simultaneously with the suspension circulation in the closed hydrodynamic circuit, the circulation in the additional circuit continues until the suspension temperature rises to the sterilization one, then the end functional product is obtained and discharged from the hydrodynamic circuit to be packed.

[0023] This method enables the production of the finished functional product - berry puree.

[0024] According to the invention the method differs in the length of the additional circuit which makes 1/2 - 1/5 the closed hydrodynamic circuit length, due to this the suspension circulating factor in the additional circuit is 2-5 times greater than in the closed one, thus ensuring additional crushing of seeds and skins.

[0024] To achieve sterilizing effect the feedstock should be held at the sterilization temperature for a predetermined time. This time can make 10-20 minutes.

[0025] According to the invention the starting sterilization temperature is to be 75-82°C.

[0026] For implementing the method there has been suggested an equipment to produce a functional product from berries in the form of puree with crushed seeds and skins; this equipment consists of a flow apparatus with an input for loading feedstock, the finished product discharge outlet with a closer, a device for air removal, a pump and a hydrodynamic baffle which form a closed circuit including a centrifugal separator which has an inlet and the first and second outlets and is installed between the hydrodynamic baffle which is connected to the centrifugal separator inlet and the flow apparatus connected with the centrifugal separator through its first outlet, a bypass channel equipped with a closer which connects the centrifugal separator second outlet and the pump sucker bypassing the flow apparatus.

[0027] The centrifugal separator can be a hydrocyclone.

[0028] The equipment implementing the invented method makes it possible to produce a functional product from berries in the form of puree which contains juice and crushed seeds and skins, the availability of polyphenols presented as a ratio of their amounts after and before processing is more than 1. Besides, the product contains water-soluble substances, their amount expressed in °Bx is 10-20% higher than in the unprocessed berries, while polyunsaturated fatty acids and vitamin E contained in the berry seeds before processing become available to the human organism.

[0029] The equipment implementing the invented method makes it possible to produce a functional product from berries belonging to the group which includes blueberry (Vaccinium myrtillus), red bilberry (Vaccinium vitisidaea) and cranberry (Oxycoccus), which contains berry juice with crushed seeds and skins, the polyunsaturated fatty acids and fat-soluble vitamins contained in the berry seeds before processing becoming available to the human organism; the content of water-soluble substances in the product expressed in °Bx is by at least 1% higher (by the product weight) than in the unprocessed berries, among them pectin makes not less than 500 mg/100 g product; mass fraction of total polyphenols is higher at least by 10%> compared to the unprocessed berries, among them anthocyanins make not less than 250 mg/100 g product; the product antiradical activity is at least 1.8 times higher than berries antiradical activity before processing; the amount of particles with characteristic size of d<50 μιη makes at least 60%, while those of d>300 μηι amount to not more than 1%.

[0030] The amount of polyunsaturated fatty acids in the functional product produced in accordance with the invention is not less than 0.7 g/100 g product.

[0031 ] The polyunsaturated fatty acids in the product are represented by a-linolenic acid (18:3) Omega-3.

[0032] Omega-3 content in the functional product produced in accordance with the invention makes at least 0.15 g/100 g product.

[0033] Vitamin E (tocopherol) content in the functional product produced in accordance with the invention makes at least 0.5 g/100 g product.

[0034] β-carotene content in the functional product produced in accordance with the invention makes at least 0.5 g/100 g product. [0035] Pectin content in the functional product produced in accordance with the invention is at least two times higher than in the unprocessed berries.

[0036] Pectin content in the functional product produced in accordance with the invention makes at least 600 mg/100 g product.

[0037] Polyphenols in the functional product produced in accordance with the invention are represented by anthocyanins.

[0038] The availability of anthocyanins is at least by 15% higher than their availability in the starting material, the anthocyanin content amounting to at least 250 mg/100 g product.

[0039] The antioxidant activity of the product produced in accordance with the invention is not less than 200 units/g.

[0040] Besides, the functional product contains not more than 0.5% particles with the characteristic size of d>300 μιη and not less than 70%) particles with the size of d<50 μηι. Brief Description of the Drawings

[0041 ] The invention will be better understood due to the following description with reference to the attached drawings in which:

[0042] Fig. 1 is a diagrammatic representation of the device according to the invention;

[0043] Fig. 2 is a device according to an embodiment of the invention;

[0044] Fig. 3 is a schematic representation of a hydrocyclone that separates the feedstock into a fraction with crushed seeds and that with uncrushed seeds;

[0045] Fig. 4 is a cross-sectional view of the hydrocyclone of fig. 3.

Best Mode for Carrying Out the Invention

[0046] The terms used in the invention description have the following meanings:

[0047] Hydrothermodynamic processing technology (hereinafter referred to as HTD- technology) is the technology of heating viscous food liquids using hydrodynamic (HDM) effect: turbulence, cavitation, particles friction against each other and against the walls of the units including those described in US Pat. No. 7,428,797. The technology provides bulk heating excluding convective surfaces of viscous materials to be processed. By passing repeatedly through the hydrodynamic zone the material particles are emulsified, effectively mixed and heated turning into pate, puree, dispersion juice, etc with stipulated degree of fragmentation. The absence of heat carrier that can overheat the feedstock allows to obtain the desired product avoiding overheating and with a long shelf-life.

[0048] HDM-baffle is a device for applying hydrodynamic effect to the feedstock. HDM- baffle can employ devices which are well-known to specialists in hydraulics and which can interrupt the laminar flow or sharply increase the flow initial turbulence. Such device can be mechanical, that is, any bluff body or an ultrasonic vibrations generator which is acoustically connected by an acoustic line to the wall of the channel through which the feedstock is pumped; it can be hydraulic (stream), for instance, in the form of an opening in the wall of the channel for pumping the main flow which opens straight into the channel and directs the exciting stream at an angle to the main flow; it can combine both mechanical and hydraulic features.

[0049] Feedstock is fresh or frozen berries, including blueberries, with small-size seeds, their removal being difficult by means of a pulping sieve, centrifuge, etc. Performing comparative analyses, blueberries were crushed to obtain pulp without access of oxygen directly before sampling, which excluded oxidation characteristic of most finished blueberry products.

[0050] "Availability" or "degree of availability" are identical values, they are determined by biochemical analyses which estimate the amount of certain substances and are performed in certified laboratories according to standard procedures.

[0051] The term "availability" can be illustrated by a walnut kernel which is unavailable for consumption while it is in the shell. The use of the original technology for making blueberry puree increases the availability of the blueberry cell content K>1. As a result of passing repeatedly the blueberry particles through HTD zone, the blueberry seeds and skins are crushed and their content in the form of blueberry cell cytoplasm and seed content get into the food liquid, thus becoming available both for analysis and for human consumption

(bioavailability).

[0052] To quantify the availability of a certain berry ingredient before and after HTD processing the value of K = mi/m ₂ is used which is determined experimentally, mi and m ₂ (mg/100 g) meaning the amount of the ingredient in berry puree and in berries processed into fresh dispersion juice immediately before sampling.

[0053] Generally speaking, the notion of availability is not identical to bioavailability but makes it possible to consider the berry puree availability for human body. This can be explained by the fact that any component in the sample is identified by chemical reactions which are similar to the biochemical reactions in the human body.

[0054] Functional product. Healthy food is getting more and more important in various countries, the core of health food being a functional product (FP). Such products influence favorably human health and are recommended as nutritional therapy or functional additives to a patient's diet. They can also serve as prophylactic agents against aging and loss of health.

[0055] The definition of the functional product accepted in Japan seems most complete and correct as it implies its every day use in the diet. The Japanese approach to functional products denies any extracts, tablets, capsules, etc even if they are made from vegetable raw materials.

[0056] Proceeding from the great experience in the nation rehabilitation after atomic bombing during the World War II and taking into account proper nutrition, this definition of the functional product is taken as a basis in this application.

[0057] The functional product in this invention is a pureed berry, including blueberry puree, is the product obtained as a result of berries processing (including blueberries) into puree-like, puree-like, cream-like substance or pulp juice with crushed seeds with different moisture content, all pathogenic microflora being destroyed due to pureeurization by HTD technology which ensures the product log shelf-life. The method of crushing and pulp, seeds and skins heat processing into a highly dispersed structure makes the product exclusive and preserves all berry healing properties plus unique components of berry seeds.

[0058] Aseptic packing. To ensure the food product shelf-life for 1-2 years, the technology of hot filling in glass jars at the pureeurization temperature of 83-95°C has been chosen as the most reliable one.

[0059] Sterilizing effect is a generally used value which is the product of heat treatment temperature by its duration. Under conventional canning technologies in continuous sterilizing production lines the sterilizing effect is achieved in conformity with the following pattern: the higher the heat treatment temperature, the shorter its duration.

[0060] Under this periodic technology of making blueberry puree the temperature grows in time or with the number of circulations, that is why sterilizing effect is determined by the sum of known coefficients multiplied by the duration of each temperature maintenance.

[0061] Sterilizing effect for berries with pH 2.8-3.5 is calculated starting from 75°C (sterilizing effect start) up to 95-98°C which is the final sterilizing effect temperature and is determined from research tables. As a rule this highest temperature is called sterilization (pureeurization) temperature.

[0062] The process of getting berry puree in accordance with the invention can be illustrated by the following examples.

[0063] Example 1. Making blueberry puree from frozen blueberries. [0064] A unit of 100 1 volume is loaded with flattened blueberries using a hatch. The feedstock consists of seeds, berry skins and juice which has been released at the initial temperature of 0°C (temperature close to defrosting). The air is removed by its forcible ousting through the unit upper part.

[0065] After removing air the hatch is sealed, the pump motor is started and the pulp is circulated along the closed circuit: pump - medium HTD separator (it operates in passive flow mode without separating fractions) - vessel - pump.

[0066] In the process of circulation the pulp is crushed and heated at the rate of 3 degrees/minute, starting from the initial temperature of 0°C up to 21°C (after approximately 7 minutes).

[0067] At this temperature blueberry skin characteristic size becomes comparable to blueberry seed size (about 1 mm).

[0068] While the circulation along the closed circuit continues, the valve is opened which feeds the material into the bypass channel, thus ensuring by centrifugal force in the separator the separation of the fraction with uncrushed seeds and its direction through the bypassing channel into the pump sucker, then into the HDM-baffle and then into the bypassing channel, and the fraction with crushed seeds and skins which after the separator moves to the vessel and then along the closed hydrodynamic circuit.

[0069] The length of the closed hydrodynamic circuit is about 5.5 m, the length of the additional circuit is about 1.4 m, that is, about 4 times shorter. Thus the multiplicity of uncrushed seeds circulations through the hydrodynamic effect zone is 4 times greater than that of the main flow, which ensures effective crushing.

[0070] While circulation in the closed and additional circuits with fractions separation continues, the feedstock temperature rises to the sterilizing effect starting temperature (80°C), then the air removal valve is opened to remove air released from berries.

[0071] During the further circulation skin and seed particles are crushed to the conditioned size of 20-50 μιιι and at the sterilization (pureeurization) temperature of 85°C the feedstock turns into blueberry puree ready for packaging; with air removed, bacterial purity ensured, the puree meets the conditions of long storage.

[0072] The temperature of 21°C in example 1 has been determined experimentally. For this in the previous experiments samples were taken at 15, 17, 19, 21 , 23°C. As the temperature was rising, the degree of skin fragmentation was increasing and at 21 -23°C the size of particles was considered sufficient for the separator efficient operation. [0073] Thus, the required growth of the suspension temperature makes At ₀ = 21°C - 0°C = 21 degree.

[0074] It should be noted that the increase of Δί ₀ by several degrees does not influence the quality of the finished product.

[0075] The decrease of Δί ₀ below 15°C causes substantial quality loss as the centrifugal separator does not ensure effective separation and crushing of seeds and large particles of berry skins.

[0076] The holding of the product at sterilizing temperature for 12 minutes improves the puree bactericidal action and lengthens its shelf-life.

[0077] Example 2.

[0078] The procedure is the same as in example 1. The sterilizing temperature is increased to 92°C. The product obtained is sterile. The additional processing manifests itself in the finished product by brown shades of the puree.

[0079] Example 3.

[0080] The procedure is the same as in example 1. The air removal from berries starts at 72°C. (The starting temperature of the sterilizing effect is also 72°C).

[0081] The finished product is nonconforming because of a great amount of air bulbs in the puree.

[0082] Example 4.

[0083] The equipment is the same. Fresh red bilberries processing starts at t ₀ = 15°C. Flattened red bilberries are fed into the unit and the procedure described in example 1 is repeated.

[0084] The optimal growth of temperature at which the bypassing channel is opened is at Δίο = 25°C, at this the suspension temperature reaches 40°C. The starting temperature of the sterilizing effect is selected experimentally and makes 76°C.

[0085] The final sterilizing temperature is 86°C and 88°C. The red bilberry puree obtained meets sanitary requirements for canned food with a long shelf-life and has pronounced bactericidal properties.

[0086] It should be noted that the lower sterilizing temperature of 86°C is preferable to the higher one of 88°C, though the degree of particle crushing at the higher temperature is higher.

[0087] In the course of the described processing the feedstock is heated to the

temperatures which together with repeated cavitation appear sufficient for destroying pathogenic microflora and removal of intercellular air. The low acid number leveling pH 3.2- 3.4 contributes to the puree preservation for at least 1-2 years, the aseptic packaging being ensured.

[0088] The functional product obtained - the blueberry puree or liquid blueberries - is a dark-blue cream-like mass with crushed seeds similar in color and scent to fresh blueberries.

[0089] Blueberry puree is a product made from natural blueberries by waste-free processing, including repeated circulation through the zone of hydrodynamic effect, accompanied by the bulk heating of preliminarily flattened blueberries, as a result the berries are heated to the sterilizing or pureeurizing temperature and crushed to homogenous state. Due to specially selected mode ("gentle" hydrodynamic processing) berry skins have been crushed to homogenous state, the characteristic size of particles being 20-50 μιη, and berry seeds have been broken due to their repeatedly passing through the HTD effect zone. The processing is performed without access of oxygen. The puree preserves blueberry unique properties.

[0090] As a result of hydrodynamic processing accompanied by heat effect, the hydrolysis has been performed, that is, the conversion of low-molecular carbohydrates into high-molecular ones (molecular chain length is increased). As distinct from conventional temperature hydrolysis, the hydrolysis performed in accordance with the invention can be termed as low- temperature or "cold hydrolysis" as it takes place at the temperatures differing by 15-20°C from the conventional one, which contributes considerably to the finished product quality.

[0091] The composition of the blueberry puree coincides with that of the natural berries it is made from.

[0092] The acidity of the puree and berries is preserved and varies within pH 3.0-3.5.

[0093] The moisture content in the puree expressed in percentage practically equals that of the feedstock and makes 82-90% depending on the berry variety and harvest time. A slight discrepancy (not more than 1%) can be explained by evaporation at the time of packing and is not taken into account.

[0094] The unique content of blueberry seeds gets into the puree and becomes available for human consumption.

[0095] The preservation of the feedstock color (dark-blue with red or purple tints) is characteristic of the product, as the properly made puree is to preserve the color of natural blueberries (or the berries it has been made from).

[0096] As forest berries from ecologically clean areas belong to organic feedstock, the puree produced from them in accordance with the invention can also be considered an organic product. [0097] Taking into account the fact that blueberries are different as far as their

biochemical and physical compositions are concerned, as most of their active substances are present in skins and seeds, all the above mentioned and used below characteristics of blueberry and other berries can refer to freshly crushed blueberries.

[0098] Thus, the functional product obtained having a long shelf-life and the feedstock of blueberries and other berries mentioned above have the same composition as well as the main characteristics (moisture content, acidity, color).

[0099] The main characteristics of puree made from blueberries, red bilberries, cranberries are:

[0100] Fully preserved total polyphenols, including chlorogenic acid (CGA) and anthocyanins, present in natural berries;

[0101] Their high availability compared to berries;

[0102] Long shelf-life due to the destruction of bacterial microflora, aseptic or quasi- aseptic sealed packaging being feasible.

[0103] Other characteristics:

[0104] 1. The amount of water-soluble carbohydrates measured in Brix degrees (°Bx) and compared to the feedstock is 1-2% higher, which is due to the following:

fiber content in puree is lower than in natural berries;

- the amount of sugars due to partial fiber "cold" hydrolysis into digestible form (as sugar and dextrins) is bigger;

- the amount of digestible pectins is increased 2-3 times, which causes the decrease of indigestible protopectins amount converting them into digestible ones due to hydrolysis, thus bringing about pronounced jelly-like properties of the puree.

[0105] 2. The content of berry seeds is transferred into the puree due to crushing most of them. The puree contains polyunsaturated fatty acid Omega-3, vitamin E (tocopherol), β- carotene, a group of sterols belonging to phyto-estrogen group.

Here the degree of K polyphenols availability (bioavailability should also be considered) is regarded as a ratio (determined experimentally) of polyphenol amount mi (mg/100 g) in the puree to their amount in natural berries m ₂ preliminarily processed into berry pulp (mg/100 g), that is, K = mi/m ₂ .

Practically in all experiments the value of K was more than 1, which testifies to the uniqueness of the puree made from blueberries or other berries by

hydrothermodynamic processing technology. [0106] Independent research has been performed in the laboratory of the Food Technology University, Kiev, using Folin-Denis method on natural blueberry feedstock and puree. Before the research natural blueberries were crushed to the size of 5-20 μιη without heating and with no oxygen present and immediately tested (control sample). Experimental samples were heated to 85-100°C using HTD technology and packed in sterile containers under quasi-aseptic conditions, then tested and compared to the experimental sample.

[0107] According to the research (see table 1) the experimental samples fully preserved the total amount of polyphenols which was in the control sample, that is, in natural blueberries. The greater amount of polyphenols after thermal processing compared to the control sample is due to their greater availability (K>1) and not to the process of synthesis which does not take place under HTD processing.

Table 1

[0108] These data confirms the assumption that polyphenols are preserved in the blueberry puree and their availability is increased by 15-20%, as, for example, for polyphenol

0.79

values in the table K = « 1.15 .

0.69

[0109] Besides, table 1 testifies to the decrease of fiber amount in the puree from 2.5% to 1.5% compared to berries, that is, by 1%, whereas the total amount of sugar increased by 0.7%, which is indicative of fiber partial conversion into sugar and dextrins. [0110] Table 1 (see also tables 4, 5) shows that the seed content unavailable in berries becomes available after applying the claimed HTD technology. In particular, the amount of polyunsaturated fatty acids makes about 1 % the puree weight. These acids include oleic acid (Omega-9), linoleic acid (Omega-6), linolenic acid (Omega-3).

[011 1] At that, the amount of PUFA Omega-3 so valuable for human health is about 300 mg/100 g puree, which makes about 20% recommended daily intake (1.5-1.8 g) of this essential acid.

[0112] It should be noted that Omega-3 is usually extracted from cod liver oil, which substantially reduces the world fish reserves. The application of the claimed method and equipment makes it possible to extract this unique PUFA from seeds crushing them in liquid with no oxygen present, thus conserving the earth natural resources.

[0113] This is also confirmed by the conversion of fat-soluble vitamin E (tocopherol) contained in seeds into the form available for human body. This vitamin requirement is compensated by 10-20%o if about 100 g puree (depending on the berry variety and maturity) is consumed daily, the daily tocopherol intake being 5 mg. The same holds true for β-carotene which is contained in the berry seed in the unavailable form and then gets into puree.

[01 14] The research performed at the independent laboratory of the Stomatology Institute, Ukrainian Academy of Science, to determine the total polyphenols, including anthocyanins, in blueberries and in the puree made from them testified to the fact that the puree contains by 25.4% more extractive substances, the amount of bioflavonoids being 16% higher, the amount of anthocyanins being 1.3 times higher (based on the weight of berries or puree, as the technology is waste-free).

[01 15] These data are given in table 2.

Experiment

Control

Na Characteristics Blueberry puree

Blueberries (A) Availability K=- (B)

1 Dry substances (%) 15,3 17,8

2 Extractive substances (% 53,6 67,2 1 ,25

based on dry weight)

3 Total bioflavonoids (% based 1,73 2,00 1 ,16

on dry weight)

4 Anthocyanins, mg/100 g 317,2 412,3 1 ,3 [01 16] The increase in the blueberry puree polyphenol and particularly anthocyanin content in the course in its production from fresh berries using HTD technology is explained by the fact that in fresh berries these substances are involved in cell structures or other substances, for instance, in sugars, which makes them unavailable in extracts produced from fresh berries. The claimed method of the puree production encourages the release of the free forms of bioflavonoids, anthocyanins, chlorogenic acid, which increases their content in the puree, this being registered by spectrophotometric analysis.

[01 17] Table 3 shows the composition of basic polyphenols in blueberries and blueberry puree, the data being obtained by using Shimadzu chromatograph, depending on the berry maturity (mg/100 g puree). (Samples 1 and 3 are berries harvested at the beginning and the end of the season; sample 2 and 4 are the puree made from these berries)

Table 3

[01 18] In particular, table 3 shows that blueberry puree is uniquely rich in anthocyanins, their content being 300-500 mg/100 g puree. It must be just anthocyanins that ensure very high antiradical activity of the blueberry puree compared to other berries and fruit.

[01 19] The results of lingonberries and red bilberry puree analyses are given in table 4.

Table 4

Sample, Sample, Availability

Characteristic Lingonberries, (mi )

86 °C 88 °C, (m ₂ ) k = m ₂ /mi Extractive substances, g/lOOg

1 13,4 15,4 15, 6 1, 16 raw material

2 Pectin, mg/100 g 170 595 629 3,7

3 Dry substances, % 19,0 18,7 18,4 -

4 Anthocyanins, mg/100 g 157,0 283,2 273,4 1 ,74

Polyphenols by Folin-Denis

5 805 972 1028 1,28 method, mg/100 g

6 Chlorogenic acid, mg / 100 g 24,8 29,2 28,9 1,17

7 PUFA Omega-3, 6, 9 mg/100 g Not available 722 863 -

8 Omega-3, mg/100 g traces 235 250 -

9 Vitamin E, mg/100 g Not available 0,72 0,76 -

10 β-carotene - 0,15 0,17 -

[0120] The amount of pectin increased 3.7 times compared to its amount in lingonberries.

[0121] The results of cranberries and cranberry puree analyses are given in table 5.

Table 5

[0122] Pectin content increased about 3.9 times compared to cranberries.

[0123] The size of berry skin and seed particles in the puree made using the method and the equipment described in the prototype are given in table 6, the data being based on blueberries. [0124] Particle sizes were measured at several temperatures, which are equivalent to the increasing multiplicity of berry particles passage through the zone of hydrodynamic processing, active turbulence and/or cavitation.

Table 6

[0125] In particular, table 6 shows that the amount of particles bigger than 300 μιη makes not more than 2%, the particles sized 150-300 μιη make not more than 5% and those of less than 150 μη make about 93% in the finished product (t° = 88°C).

[0126] The degree of crushing corresponds to the requirements of Ukrainian State Standards 4082-2001 "Puree-like canned fruit for baby food. Specifications for homogenized products for baby food". In accordance with them the amount of particles with d<150 μπι is to be not less than 70%, d>300 μηι - not more than 7%.

[0127] The characteristic sizes of crushed blueberries ensured by the invention are given in table 7.

Table 7

[0128] Table 7 shows that in the finished product (t° = 88°C) there are practically no particles with d>300 μηα (>1%), particles sized 150-300 μιη amount to not more than 2%, particles sized less than 150 μιη make at least 98%.

[0129] The analysis with the help of an electronic microscope showed that the

characteristic diameter of particles is 80-100 μιη in berry purees produced by the method described in the prototype (table 6) and 20-50 μηι in berry purees produced by the claimed method (table 7).

[0130] Given the repeatability of the results with cranberries and red bilberries, it can be concluded that the functional product of the berry puree has a characteristic spectrum of particles crushing to d<150 μηι amounting to 97%, among them the particles with d<50 making not less than 60%.

[0131] It is probably because of this that the berry purees shown in tables 1-5 and made in accordance with the claimed method and equipment have pronounced jelly-like properties.

[0132] These properties are due to the presence of considerable amounts of pectin substances which form a three-dimensional ordered structure.

[0133] As a rule, blueberries, cranberries and red bilberries contain from 0.5% to 0.9% protopectin. As a result of low-temperature hydrolysis taking place at low temperatures owing to cavitation and turbulence, a great amount of protopectin is converted into pectin (water- soluble form) which is available to human organism.

[0134] As a result, the amount of water-soluble and extracted substances increases by 1- 2% which corresponds to 10-20% based on the puree dry weight. About half of this amount consists of pectin substances, the other half - of simple sugars and dextrins obtained by fiber hydrolysis, whereas blueberry or other berry juice made from fresh or frozen berries has a ten times lower viscosity than the viscosity of purees made from these berries using the invention.

[0135] Berry juice flows easily down a vertical surface, spreads on a horizontal surface like any Newtonian liquid. Drop formation in such juice is similar to water drop formation.

[0136] As distinct from juice, the berry puree is characterized by high degree of greasiness and adhesiveness. In particular, the puree does not flow down from a vertical surface and does not spread on a horizontal surface under attractive force.

[0137] As it is difficult to describe the quantitative characteristics of jelly-like puree properties, only their qualitative characteristics are mentioned.

[0138] Below are the results of the research aimed at determining the antioxidant activity (AA) of blueberries and the puree made from them. It has been found out that the puree AA is considerably higher than the blueberries AA (table 8).

Table 8

[0139] Blueberry puree biochemical characteristics (average of three values)

T e ant ox ant propert es o ue err es an o the puree made from them have been estimated according to three characteristics: antiradical activity (using DPPH free radical), anti superoxide activity (inactivation of superoxide antiradical), chelating activity (binding of iron ions Fe ²⁺ ).

[0141] It has been noted that blueberry puree antiradical and chelating properties exceed those of all known antioxidants, among them grape leaves, which are a powerful source of polyphenols, are 2.5 times inferior to blueberry puree.

[0142] Thus, on the basis of the above materials and numerous independent studies conducted by the applicant we can state that the blueberry puree is a unique natural antioxidant product which can be attributed to functional products.

[0143] It is important to note that due to HTD processing the biochemical activity of natural blueberries rises 1.5-2 times, blueberry itself being a powerful natural antioxidant. At that, the claimed method and equipment improve the unique properties of fresh natural berries, preserving them in canned form for 1 -2 years (which is quite unattainable with fresh berries).

[0144] It should be mentioned that the blueberry puree made by the method described in the prototype does not provide such a high degree of anthocyanin availability. Consequently, the antioxidant activity of such product is considerably lower than that of the product made by the method in accordance with the invention.

[0145] Table 8 shows, for example, that the antiradical activity (ARA) of the puree made according to the invention is about 2 times higher than that of the blueberries the puree is made from (K = 2.06). [0146] At the same time, the ARA of the blueberry puree made according to the prototype method is only 1 .3 times higher than the ARA of the berries (Kl = 1.29).

[0147] Blueberry puree is known for its positive prophylactic and therapeutic effect in treating such serious diseases hepatitis, dysbacteriosis (dysbiosis), vascular diseases, including hepatitis eye-lesion, certain types of cancer, genitourinary infections, such as trichomonads, lamblias, etc.

[0148] Owing to its fine-dispersed crushing (nanocrushing) liquid blueberries are easily digested ensuring transportation of polyphenols, antioxidants, including anthocyanins, fiber, pectin, fructose, glucose, carotinoids, tocopherol, Omega-3, etc, to the affected organ, which testifies to the product functionality.

[0149] Blueberry, cranberry and red bilberry samples have been studied for their antibacterial activity. The highest activity has been determined in red bilberry and cranberry samples. The research was conducted first on "pure" culture of bacteria, then on the bacteria of clinical cultures. The latter has been of particular interest as these bacteria acquired immunity to most standard medicines. As distinct from chemicals, berry puree does not oppress prebiotic, or "useful" microflora.

[0150] In accordance with WHO recommendations to assess the product biological activity the following test-strains have been used: Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Basillus subtilis ATCC 6633, Proteus vulgaris ATCC 4636, Candida albicans ATCC 885 /653.

[0151] Microbial suspension was made using Densi-La-Meter device (made by PLIVA - Lachema, Czech Republic; 540 nm wave length). The suspension was prepared according to the instruction the device is supplied with and the information letter on the innovations in the health care system # 163-2006 "Standards in Microbial Suspension Preparation", Kiev. Cultures were synchronized for 2 hours at low (4°C) temperature. The microbial load was 107 microbe cells per 1 ml medium. 18-24-hour microorganism culture was used. The tests were performed using Mueller-Hinton agar (PA "Life-Giving Medium", Dagestan, XI, 2013 expiration date).

[0152] Preparation diffusion in agar was performed by "wells" method. Preparation antibacterial activity was determined on two layers of solid medium in Petri dishes. In the lower layer use was made of "hungry" unseeded media (agar-agar, water, salts). The lower layer was a 10 mm thick lining with 3-6 vertically installed stainless steel thin-walled cylinders (8 mm diameter, 10 mm high). The upper layer was poured around cylinders, it consisted of life-giving agar medium, melted and cooled to 40°C into which an appropriate standard of test- strain daily culture was introduced. Preliminarily this layer was carefully mixed to form a homogenous mass. After the mass stiffened, the cylinders were removed with sterile forceps and the holes were filled with the material to be tested considering its volume (0.3 ml).

[0153] The medium volume of the upper laver varied from 14 to 16 ml. The dishes were dried for 30-40 minutes at room temperature and then put into a thermostat for 18-24 hours.

[0154] The following criteria have been used to assess new antibacterial substances and to study strains resistance to antibiotics:

absence of microorganism growth-inhibition zones around holes as well as growth- inhibition zones less than 10 mm testify to microorganism insensibility to the preparation in the hole or to antibiotic concentration;

[0155] 10-15 mm diameter growth-inhibition zones testify to the culture low sensibility to the antibacterial substance concentration;

15-25 mm diameter growth-inhibition zones testify to the microorganism sensibility to the drug;

over 25 mm diameter growth- inhibition zones testify to the microorganism high sensibility to the drug.

[0156] The results of the research on the samples antibacterial activity are given in tables 9 and 10.

Table 9. Samples antibacterial activity

Growth-inhibition zone diameters, mm

Number of experiments n = 3

Lingonberries Lingonberries Lingonberries Cranberry 1 2 3

Staphylococcus 25, 25, 25 25, 26, 26 25, 26, 26 27, 26, 25 aureusATCC 25923

Escherichia coli ATCC 24, 23, 23 24, 24,24 24, 24, 25 25, 24, 25 25922

Pseudomonas 18, 19, 19 18, 18, 19 19, 18, 19 20, 21, 21 aeruginosa ATCC 27853

Proteus Vulgaris ATCC 17, 17, 17 16, 17, 16 17, 17, 17 16, 17, 16 4636 Basillus subtilis ATCC 6633 27, 28, 28 28, 28, 29 27, 28, 28 29, 28, 28

Candida albicans ATCC 17, 18, 17 17, 17, 18 19, 19, 18 18, 19, 17 653/885

Staphylococcus 25, 25,26 25, 26, 25 24,25,25 25, 24, 25 aureus(clinical)

Staphylococcus aureus 26, 25, 25 24, 25, 25 25,25,25 26, 26, 27 (clinical)

Escherichia coli 24, 25, 24 24, 24, 24 24, 25, 23 23, 23, 24 (clinical)

Pseudomonas aeruginosa 15, 16, 16 16, 17, 17 17, 16, 17 16, 17, 17 (clinical)

Enterobacter aerogenes 21,20, 22 22,21,21 22,21,21 22,21,22 (clinical)

Enterobacter aerogenes 20,21,20 22,21,21 22, 22, 22 23, 22, 22 (clinical)

Klebsiella pneumonia 20,21,20 21,22,21 20, 20, 20 21,20,21 (clinical)

Edwardsiella tarda 22,21,22 21,22,21 20, 22,21 21,20,21 (clinical)

Citrobacter freundii 25, 26, 25 25, 26, 25 25, 25, 25 25, 26, 26 (clinical)

Proteus mirabilis 18, 17, 17 16, 16, 16 17, 16, 17 16, 17, 16 (clinical)

Lactococcus sp 15, 16, 17 16, 17, 16 15, 16, 16 15, 16, 17

Candida albicans 15, 16, 15 15, 16, 16 16, 15, 16 15, 16, 16 (clinical)

Enterobacter aerogenes 20,21,20 22,21,21 22, 22, 22 23, 22, 22 (clinical)

Klebsiella pneumonia 20,21,20 21,22,21 20, 20, 20 21,20,21 (clinical)

Edwardsiella tarda 22,21,22 21,22,21 20, 22, 21 21,20,21 (clinical)

Citrobacter freundii 25, 26, 25 25, 26, 25 25, 25, 25 25, 26, 26 (clinical)

Proteus mirabilis 18, 17, 17 16, 16, 16 17, 16, 17 16, 17, 16 (clinical)

Lactococcus sp 15, 16, 17 16, 17, 16 15, 16, 16 15, 16, 17

Candida albicans 15, 16, 15 15, 16, 16 16, 15, 16 15, 16, 16 (clinical)

Table 10. Antibacterial activity of samples after 10-fold saline dilution

Growth-inhibition zone diameter, mm

Number of experiments n = 3

Lingonberries 1 Lingonberries 2 Lingonberries Cranberryl : 10 1 : 10 1 : 10 1 : 10

Staphylococcus 20, 19, 19 19., 18, 19 19, 19, 20 20, 20, 19 aureusATCC 25923

Escherichia coli ATCC 17, 18, 18 17, 18, 18 17, 18, 17 17, 17, 19 25922

Pseudomonas 1 5, 16, 15 15, 15, 16 16, 15, 14 15, 15, 16 aeruginosaATCC

27853

Proteus Vulgaris ATCC 15, 15, 14 14, 14, 14 14, 15, 14 15, 14, 15 4636

Basillus subtilis ATCC 20, 21 , 20 20, 21 , 21 20, 20, 21 22, 21, 20 6633

Candida 14, 14, 15 14, 14, 14 15, 14, 15 15, 15, 15 albicans ATCC

653/885

Staphylococcus 19, 18, 18 18, 18, 19 17, 18, 19 19, 18, 19 aureus(clinical)

Staphylococcus aureus 18, 18, 18 17, 18, 17 17, 18, 18 19, 18, 18 (clinical) Escherichia coli 17, 18, 17 18, 17, 17 17, 18, 17 18, 18, 16 (clinical)

Pseudomonas 14, 15, 15 16, 15, 16 14, 15, 15 16, 15, 15 aeruginosa

(ΚΠΙΗΪΗΗΗΗ)

Enterobacter aerogenes 15, 14, 14 14, 14, 14 15, 15, 15 15, 15, 14

(KjliHlHHHH)

Enterobacter aerogenes 16, 16, 15 18, 17, 16 16, 14, 16 14, 14, 16

( jliHiqHHH)

Klebsiella pneumonia 15, 15, 15 14, 14, 15 16, 16, 15 15, 15, 14

(KjliHlHHHH)

Edwardsiella tarda 16, 16, 17 15, 15, 16 17, 15, 15 15, 16, 16

(KjliHlHHHH)

Citrobacter freundii 16, 1 5, 16 16, 18, 16 17, 1 5, 16 1 5, 16, 16 (clinical)

Proteus mirabilis 14, 14, 14 15, 15, 15 16, 14, 15 15, 14, 14 (clinical)

Lactococcus sp growing growing growing growing

Candida albicans growing growing growing growing (clinical)

Candida 14, 14, 15 14, 14, 14 15, 14, 15 15, 15, 15 albicans ATCC

653/885

Staphylococcus 19, 18, 18 18, 18, 19 17, 18, 19 19, 18, 19 aureus(clinical)

Staphylococcus aureus 18, 18, 18 17, 18, 17 17, 18, 18 19, 18, 18 (clinical)

Escherichia coli 17, 18, 17 18, 17, 17 17, 18, 17 18, 18, 16 (clinical)

Pseudomonas 14, 1 5, 15 16, 15, 16 14, 15, 15 16, 15, 15 aeruginosa

(clinical) Enterobacter aerogenes 15, 14, 14 14, 14, 14 15, 15, 15 15, 15, 14 (clinical)

Enterobacter aerogenes 16, 16, 15 18, 17, 16 16, 14, 16 14, 14, 16 (clinical)

Klebsiella pneumonia 15, 15, 15 14, 14, 15 16, 16, 15 15, 15, 14 (clinical)

Edwardsiella tarda 16, 16, 17 15, 15, 16 17, 15, 15 15, 16, 16 (clinical)

Citrobacter freundii 16, 15, 16 16, 18, 16 17, 15, 16 15, 16, 16 (clinical)

Proteus mirabilis 14, 14, 14 15, 15, 15 16, 14, 15 15, 14, 14 (clinical)

Lactococcus sp growing growing growing growing

Candida albicans growing growing growing growing (clinical)

[0157] As a result of the research it can be stated that red bilberry and cranberry samples possess antibacterial properties towards various microorganisms and yeast-like fungi, the anti microbial activity being preserved at 10-fold dilution.

Industrial Applicability

[0158] The invention in any embodiment of the inventor's idea can be implemented industrially using simple non-standard equipment. The invention is designed for food industry usage for the production of such puree-like products from vegetable raw materials which are enriched with fats based on unsaturated fatty acids, fat-soluble vitamins, polyphenols, including anthocyanins with high antiradical activity in cases the desired product is to meet high requirement concerning its taste and functional properties as well as long shelf-life.