TECHNICAL FIELD

The present invention concerns a method for preparing preservatives in the food industry.

BACKGROUND ART

Many food products require the addition of chemicals to prevent deterioration due to the action of micro- organisms or oxidation by the oxygen contained in the air. Said chemicals are generally indicated as food preservatives.

Few food preservatives are truly harmless. The majority of food preservatives can have side effects if certain consumption limits are exceeded. These food preservatives include sulphites, nitrites and nitrates.

For a correct evaluation of the toxicity of these food preservatives, it is important to remember, for example, that nitrates and nitrites are converted by the organism that consumes them into nitrosamines , which are often involved in potentially carcinogenic processes. Furthermore, sulphites, which are often added to foodstuffs in the form of sulphur dioxide (S0 ₂ ) , in sensitive persons can trigger allergies which manifest themselves in respiratory disorders, and are in any case toxic for everyone if consumed in high quantities.

In the food industry, the need is therefore strongly felt for preservatives that are effective in preventing the deterioration of foodstuffs and at the same time are free from toxicological contraindications .

The Applicants have devised a method for producing food preservatives starting from extracts and molecules from appropriate vegetable matrices so as to replace the molecules produced by chemical synthesis such as, for example, the sulphites, nitrites and nitrates currently used massively in the agri-food industry. The preservatives obtained by the method of the present invention can be used for the production, for example, of enological products, fruit juices, beer, and cured and/or cooked meat based products.

For a correct understanding of the invention, a brief explanation of clustered water is provided below. Studies based on Quantum Electro-Dynamics (Del Giudice E. , Gualdi C. , Mangano G. , Mele R. , Miele G. , Preparata G. , "Neutron stars and the coherent nuclear interaction", International Journal of Modern Physics D , 531, (1995); Del Giudice E., Galimberti A., Gamberale I., Preparata G. , "Electrodynamical Coherence in Water: a possible origin of the tetrahedral coordination", Modern Physics Letters B9, 953, (1995)) have explained the long range correlations and the formation of clusters in water via a description based on a system of interacting dipoles. According to these studies, in the transition of the physical and energy states of water, another actor exists which is the magnetic field. It has been hypothesised that in the transitions from one level to another, the atom changes the form and arrangement of its charges, emitting at the frequency ω = ΔΕ/¾. When these atoms have the same frequency and the same wavelength, they begin to exchange electromagnetic messages, as in the binary language 1 and 0 of computer science, so as to synchronise (phase) their oscillations and/or transitions. The regions in which all this occurs are called "coherence domains" and have the dimensions of -1000A.

The molecules within the coherence domains oscillate completely in phase and assume variable energy states, superior to the minimum energy states .

The magnetic field that acts mainly to trigger the coherence domains can take many forms: it can be a static field of a permanent magnet, it can be a dynamic field of an electromagnetic wave, with frequency sufficient to resonate the electrons or, considering the quantum field (infinitely small and instantaneous) , it can be the information of other coherence domains of other water which is already excited.

Examining the water, when it is in its state of natural equilibrium, its electric dipoles are in an incoherent regime, i.e. they are arranged in a disorderly manner with electrostatic type bonds.

If the water is immersed in a magnetic field greater than the critical value corresponding to the natural equilibrium, the dipoles are oriented in a coherent regime; in this situation the dipoles oscillate in phase with one another. In situations of dynamic equilibrium, the coherence domains are positioned alongside one another, so that the coherences are linked together. The "size" of the coherence domain is equal to the wavelength of this coherent field and, in the case of water, corresponds to 1/10 micron.

For every given temperature value, and therefore for a given number of collisions with the molecules of the environment, a fraction of the water molecules "loses its rhythm" and feeds a non-coherent fraction which, like a dense gas, circulates in the interstices between the coherence domains. The remaining 60% of water is not dominated by the coherent magnetic field and represents the non-coherent fraction which behaves like a physical system that follows the laws of gases.

The coherence domains can be displayed as islands immersed in a sea of non- coherent liquid water. The fraction of coherent water is organised in complex structures which simulate the hydrogen bond and form magnetic structures; these structures are able to interact with extremely weak electromagnetic signals and transport the electromagnetic information. Therefore, the coherent part of water can receive and transport electromagnetic information, while the non-coherent part represents the solvent of the ions and elements fundamental for functioning of the cell. Within these fields, the water assumes for the most part a hexagonal structure. The capacity of the water to form clusters is used to incorporate extraneous molecules, for example sugars, salts, proteins, acids or active ingredients.

The water molecules imprison the extraneous molecule and surround it, forming a shell or a niche and create a "copy"; even when this extraneous molecule is completely destructured or filtered and therefore the niche is empty, the clusters appear to maintain its form, its perfect "footprint" and record its information in terms of vibrations.

Short and long distance interactions therefore exist which bind the water molecules into coherence domains and allow them to preserve the structure they had in the presence of a solute, even after the disappearance of the latter.

Here and below, by clustered water we mean water that has been subjected to the action of an electromagnetic field and which has the characteristics described above.

DISCLOSURE OF INVENTION

The subject of the present invention is a method for preparing preservatives in the food industry comprising in an operating sequence the steps of:

ultrasound extraction, in which a plant species with antioxidant and antimicrobial properties is added to clustered water and subjected to sonication;

- separation, in which a phytoextract mixture coming from the extraction stage is subjected to an extraction with supercritical fluid chromatography; - clustering, in which the phytoextracts obtained from the separation stage are mixed with water and the mixture formed is subjected to a clustering process by means of the action of an electromagnetic field.

BEST MODE FOR CARRYING OUT THE INVENTION

Preferably, the clustered water used in the ultrasound extraction stage and produced in the clustering step has a pH that ranges from 5.0 to 8.5 and a redox potential that ranges from 5 to 40 mV.

Preferably, the clustered water used in the ultrasound extraction step is obtained by subjecting a quantity of double-distilled water with a temperature ranging from 5 to 15°C and having a pH ranging from 7.0 to 8.0 to a direct current having a voltage ranging from 5 to 27 volts for an amount of time ranging from 0.5 to 3.5 hours.

Preferably, in said ultrasound extraction step the sonication has a frequency ranging from 100 to 300 KHz.

Preferably, in said separation stage the supercritical fluid is carbon dioxide used at a temperature ranging from 31 to 35 °C and at a pressure ranging from 70 to 80 atm. Preferably, the plant species added to clustered water is comprised in the group consisting of Vitis Vinifera; Rosmarinus Officinalis; Malpighia Punicifolia; Citrus Limonum; Raphanus Niger, Curcumin, Devil's Claw; Poligonum Cospidatum, Panax Ginseng, Gingko Biloba and White Willow.

For a better understanding of the present invention, implementation examples for illustrative non-limiting purposes are given below. PREPARATION OF PRESERVATIVE FOR CURED PORK

- Water clustering - A cylindrical toughened glass container comprises 1 litre of double-distilled water having a temperature of 9°C and a pH of 7.0. Two graphite electrodes are connected to the container, arranged at a height equal to 1/3 and 2/3 of the water level starting from the base of the container. The two electrodes are connected to a direct current delivery device and subjected to a voltage of 15 volts for a period of 2 hours. At the end of the 2 hours of delivery, the water inside the container has a pH value of 8 and a redox potential of 12 mV.

- Ultrasound extraction -

The clustered water coming from the clustering stage is poured into a cylindrical stainless steel tank. In the tank, 4 digital multifrequency ultrasound generators are radially installed capable of emitting ultrasounds with a bandwidth oscillating between 20 and 350 KHZ. In particular, the ultrasound generators are positioned at a height equal to 2/3 of the water level starting from the base of the tank. Inside the cylinder there is a motorised propeller stirrer.

50 g of Rosmarinus Officinalis leaf with dimension of 2 mm are placed inside the tank.

The mixture thus constituted is stirred, at a frequency of 100 Hz, for 15 minutes and subsequently the ultrasound generators are activated at a frequency of 250 KHZ for 2 hours.

Once the sonication stage has been completed, the exhausted vegetable raw material is eliminated, while the liquid portion consisting of clustered water and phytoextract is collected ready for the next separation stage.

- Chromatographic separation with supercritical fluid - Supercritical carbon dioxide at a temperature of 35 °C and a pressure of 75 atm is used as the supercritical fluid. The extraction column is filled with bauxite having a diameter of 1 mm and with zeolite having a diameter of 2 mm. Furthermore, the extraction column is split into a feed section, a rectification section (portion of the column above the feed section) and an exhaustion section (portion of the column below the feed section) .

The fractionation occurs by temperature gradient. The column is equipped with a differentiated heating system throughout the height, where fractionation is performed by temperature gradient with temperatures ranging from 25 to 35 °C and increasing with the height.

In the exhaustion section where the temperature ranges from 25 to 35°C the compounds to be fractionated are solubilised; in the rectification section, the temperature ranges from 8 to 15 °C so as to drastically reduce the solubility of one or two compounds; the latter will be released by the C0 ₂ -SC and will undergo an internal reflux, while the more soluble compound will become increasingly concentrated in the extract. The lightest portion of phytoextract is collected from the top of the extraction column. Said light portion of phytoextract is subsequently subjected to the clustering stage. - Clustering -

50 ml of the light portion of phytoextract obtained from the separation stage are mixed with 950 ml of double-distilled water at a temperature of 10 °C and having a pH of 5.50. The solution thus obtained is subjected to a further clustering and final stabilisation process.

The solution is then placed in a cylindrical toughened glass container identical to the one described for clustering of the water. Two graphite electrodes are applied in the container, arranged at a height of the solution equal to 2/3 starting from the base of the container. The two electrodes are connected to a direct current delivery device and subjected to a voltage of 20 volts for a period of 2 hours. At the end of the treatment, the solution has a pH value of 8 and a redox potential value of 10 mV.

One litre of the resulting solution has the following composition: 950 ml of clustered water; 25 ml of Rosmarinic Acid; 10 ml of Erodithiol; 4 ml of Ampelotin; 3 ml of Naringenin; 3 ml of Peonidin; 3 ml of Malvidin; 2 ml of Apigenin.

- Test on preserving power for cured pork -

The resulting solution was tested as a preservative for cured meats.

In particular, 1 ml of the above solution per 1000 g of cured meat was used. As a comparison, the traditional preservative used was a mixture consisting of 2 g of Potassium Nitrate (E 252) and 1 g of Potassium Nitrite (E 249) per 1000 g of the same cured meat .

After 30 days, a sample of 100 g of each of the two cured meats was analysed to verify the presence of bacteria.

A further comparison example consists in repetition of the procedure described above, the sole difference being that the ultrasound extraction stage is performed in the presence of double-distilled water not previously subjected to the clustering stage. The ultrasound extraction is much improved in the presence of clustered water, and this improvement necessarily affects the preserving power of the product derived . demonstrate the above, it was experimentally verified that ultrasound extraction stage of the above example (ultrasound extraction in the presence of clustered water) produces a phytoextract concentration equal to 0.21 Kg/m ³ , whereas repeating the same procedure with double-distilled but non-clustered water, the phytoextract concentration is lowered to 0.13 Kg/m ³ .

Table I shows the bacterial load values in CFU/g recorded on the cured meat comprising the food preservative produced by the present invention (Cured meat inv.) , on the cured meat comprising the traditional food preservative (Cured meat pack 1) and on the cured meat comprising the food preservative produced without using clustered water in the ultrasound extraction stage (Cured meat pack 2) .

TABLE I

PREPARATION OF PRESERVATIVE FOR ENOLOGICAL PRODUCTS

- Water clustering -

A cylindrical toughened glass container comprises 1 litre of double-distilled water having a temperature of 10°C and a pH of 7.5. Two graphite electrodes are connected to the container, arranged at a height equal to 1/3 and 2/3 of the water level starting from the base of the container. The two electrodes are connected to a direct current delivery device and subjected to a voltage of 13 volts for a period of 2 hours. At the end of the 2 hours of delivery, the water inside the container has a pH value of 8 and a redox potential of 13 mV.

- Ultrasound extraction - The clustered water coming from the clustering stage is poured into a tank identical to the one already described in the corresponding stage for the preparation of preservatives for cured meat . 50 g of Rosmarinus Officinalis leaf with dimension of 2 mm and 40 g of Vitis Vinifera seeds with dimension of 1 mm are placed inside the tank.

The mixture thus constituted is stirred at a frequency of 100 Hz for 25 minutes, after which the ultrasound generators are activated at a frequency of 270 KHZ for 3.0 hours.

Once the sonication stage has been completed, the exhausted vegetable raw material is eliminated and the liquid portion consisting of clustered water and phytoextract is collected ready for the next separation stage.

- Chromatographic separation with supercritical fluid - Supercritical carbon dioxide at a temperature of 34 °C and a pressure of 77 atm is used as the supercritical fluid.

The stage uses an extraction column and a method identical to those described above for the corresponding stage in the preparation of preservatives for cured meat. The lightest portion of phytoextract is collected from the top of the extraction column. Said light portion of phytoextract subsequently undergoes the clustering stage.

- Clustering - 100 ml of the light portion of phytoextract obtained from the separation stage are mixed with 900 ml of double-distilled water at a temperature of 10°C and having a pH of 6.50.

The solution thus obtained is subjected to a further clustering and final stabilisation process.

The solution is then placed in a cylindrical toughened glass container identical to the one described for clustering of the water. Two graphite electrodes are applied in the container, arranged at a height of the solution equal to 2/3 starting from the base of the container. The two electrodes are connected to a direct current delivery device and subjected to a voltage of 25 volts for a period of 2 hours. At the end of the treatment, the solution has a pH value of 8 and a redox potential value of 12 mV.

One litre of the resulting solution has the following composition: 900 ml of clustered water; 3 ml of Afzelechin; 6 ml of Apigenin; 8 ml of Ampelotin; 6 ml of Cyanidin; 8 ml of Peonidin; 9 ml of Malvidin; 4 ml of Anthocyanins ; 27 ml of Procyanidins ; 25 ml of Rosmarinic Acid; 4 ml of Thiamin.

- Test of preservation power on enological products -

The resulting solution was tested as a preservative for enological products.

In particular, 3 ml of solution were used per 1000 ml of wine. For comparison, sulphur dioxide was used as a traditional preservative in a quantity of 36 mg per 1000 ml of the same wine .

A further comparison example consists in repetition of the procedure described above, the sole difference being that the ultrasound extraction stage takes place in the presence of double-distilled water not subjected to the clustering stage. The ultrasound extraction is much improved in the presence of clustered water, and this improvement necessarily affects the preserving power of the product derived.

To demonstrate the above, it was experimentally verified that the ultrasound extraction stage of the above example (ultrasound extraction in the presence of clustered water) produces a phytoextract concentration equal to 0.20 *Kg/m ³ , whereas repeating the same procedure with double-distilled but non-clustered water, the phytoextract concentration is lowered to 0.12 Kg/m ³ .

After 210 days, a sample of 50 ml of each of the two wines was analysed to verify the presence of the compounds shown in Table II and considered the main indicators of correct vinification.

Table II shows the values in mg/litre of the above-mentioned compounds both for the wine treated with the preservative obtained by the method of the present invention (Wine inv.), and for the wine treated with the traditional preservative sulphur dioxide (Wine pack 1) and for the wine treated with the preservative obtained by means of an ultrasound extraction stage in the presence of non-clustered double-distilled water (Wine pack 2) .

TABLE II

The values of the parameters given in Table II show that the wine comprising the preservative prepared with the method of the present invention has a vinification in terms of alcoholic fermentation and malolactic fermentation similar to if not better than that of wines comprising sulphur dioxide as a preservative. In particular, the wine comprising the preservative obtained with the method of the present invention with respect to the comparison wine shows a substantial similarity as regards the parameters of shikimic acid (index of antioxidant qualities of the wine) and tartaric acid (index of freshness and vivacity of the wine) , whereas it shows a drastic reduction in the levels of sulphur dioxide, thus demonstrating the possibility of being able to eliminate it completely, maintaining or even improving the other qualitative parameters.

Lastly, a lower quantity of malic acid shows an improvement in malolactic fermentation and, therefore, a "softer" wine, whereas a higher quantity of lactic acid shows a lower acidity.

As appears evident from the results given in Tables I and II, the preservatives prepared with the method of the present invention are able to guarantee preservation of the food product to which they are applied and, at the same time, do not entail the problems of toxicity of the food preservatives.

Lastly it should be highlighted that the presence of clustered water in the ultrasound extraction stage is able to guarantee a more effective extraction power, therefore providing a product with a higher preserving power.