FI64272C - FOERFARANDE FOER FRAMSTAELLNING AV NAERINGSMEDEL INNEHAOLLANDEETT I MAGTARMKANALEN AKTIVERBAT ANTIBAKTERIELLT SYSTEM - Google Patents

Description

1 64272

The present invention relates to a process for the preparation of novel nutrients comprising a gastrointestinal activating antibacterial system.

It is known that children in particular can develop severe gastrointestinal infections, mainly due to infections caused by E.coli and various Salmonella Species. Thus, previously known methods for eliminating these infections have involved administering a therapeutic amount of antibiotics at the risk of forming an antibiotic-resistant bacterial strain and so on.

It has now surprisingly been found possible to avoid the latter disadvantages and to prevent and / or eliminate such bacterial infections by means of the present invention, which is characterized by the addition of an antibacterial system containing at least 1 mg / kg of nutrient (50-5000 U / kg) to known nutrient mixtures. nutrient) lactoperoxidase, at least 16 ppm thiocyanate calculated as sodium thiocyanate and at least 21 ppm solid, water-soluble peroxide donor which is alkali metal carbonate, alkaline earth peroxide or urea peroxide, calculated as sodium percarbonate.

The molar ratio of peroxide donor to thiocyanate is preferably less than 4: 1, preferably 1-2: 1.

In a preferred embodiment, thiocyanate .60-3500, preferably 160-1750 ppm / kg of nutrient calculated as NaSCN, peroxide donor in the amount of 2 64272 210-4000, preferably 210-2000 ppm / kg of nutrient calculated as sodium percarbonate and lactoperoxidase 10-200 are added. , preferably 10-100 mg / kg of nutrient.

The amount of thiocyanate is chosen so that its concentration in the digestive tract is 0.1, preferably 0.2-0.4 mM, whereby the concentration must not exceed the toxic concentrations (10 mM). The amount of peroxide donor is selected to give an equimolar amount, i. so that the concentration of H 2 Cl 2 formed reaches 0.1, preferably 0.2 to 0.4 nM. The amount of lactoperoxidase depends on the activity of the enzyme, but assuming that 1 mg of enzyme contains 50 units (U), at least 1 mg of enzyme per liter of gastric fluid must be present.

The concentration of thiocyanate in the solution is thus 0.1, preferably 0.2-0.4 mM, the concentration of the peroxide donor is such that the concentration of hydrogen peroxide is 0.1, preferably 0.2-0.4 mM and the concentration of lactoperoxidase is 1 mg / 1 (50 U / l). The latter may vary but must be 1-2 mg / l (50-100 U / l).

One unit of lactoperoxidase is the amount of lactoperoxidase-hih, which forms 1 mg of pyrogallin from pyrogallol in 80 seconds at pH 6.0 and 20 ° C.

The amount of lactoperoxidase in bovine whey (unpasteurized) or ultrafiltered whey can, of course, vary, the amount of lactoperoxidase reported in the literature is about 30 mg / l, while the amount of lactoperoxidase in human milk is reported to be 1/30 of bovine milk.

The invention is described in more detail below by way of example.

£ Šimčík

The broth for young children was made from the following i. St a a i.neks 3 64272 50, 3 kg of unsalted whey powder.

19.0 kg skimmed milk powder (Z 100 mg LP / kg) 14.0 kg fat blend 8.0 ke lactose 7.5 kg steamed rice powder __1.0 kg vitamins and minerals 100.0 kg 21 g sodium percarbonate 16 g sodium thiocyanate

The ingredients were thoroughly mixed in a blender. The dry broth mixture is dissolved in water at a temperature of about 30-3 5 ° · "; to a 10-15% aqueous solution (10-15% dry matter) and administered in an amount of about 0.2 liters 4-5 times a day.

The amount of sodium percarbonate in such broth may vary between 210-420 ppm and sodium thiocyanate between 160-320, the amounts preferably being equinolar.

The sodium percarbonate used in the above example is protected against the purple reaction by coating the granulate with a layer of a polymer which is soluble or permeable only to the intestinal tract.

Such protective polymers include waxes, cellulose gun tantalum, phthalate, and polymers that dissolve or permeate in the intestinal region. In addition, a dual coating may be used if the slurry to be performed prior to administration would result in an environment in which the polymer dissolves.

Biological effect

Four test tubes containing 10 ml of whey with 21 ppm (0.25 mM) sodium thiocyanate were incubated at 30 ° C inoculated with Ps. Fluorescence EF 1998. One of the tubes was a control. Sodium percarbonate was added to the other three test tubes in amounts corresponding to 0.1, 0.2 and 0.3 mM U 00. The number of bacteria was determined at inoculation and 2 hours after inoculation. LactoDeroxidase was present at 5 ug / ml. The results are given in Table 1 below.

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table 1

Sodium percarbonate Number of bacteria / ml in an amount corresponding to _. ~ mM H 2 O 2 ° for 2 hours

Control (0.0 mM) 2.1 · 106 2.1 · 106 0.1 mM 2.1-106 1.8-105 0.2 mM 2.1 - 106 2.8 · 103 0. 3 mM 2 , 1-106 2.6-103

As can be seen from this, a strong improvement in antibacterial activity was obtained when 0.2 mM H 2 O 2 or more is present. However, the addition of as little as 0.1 mM H 2 O 2 gives a strong bactericidal effect.

Ps fluorescence EF 1998 was incubated at + 30 ° C in two test tubes 1) and 2), tube 1) containing 1 g of commercial milk replacer with 15% ultrafiltered whey powder, 57% whey powder, 8% fat, 16% animal protein and 4% of vitamins and minerals in 10 ml of water containing 50 ug of lactoperoxidases and 27.5 ppm of sodium percarbonate was added thereto, and tube 2) contained the same amount of milk replacer (50 ug of lactoperoxidase) and having 21 ppm of sodium thiocyanate and 27.5 ppm of ppm sodium percarbonate I isatri in.

The number of bacteria in the test tubes is given in Table 2 below, which gives the results for 0, 2, 4 and 6 hours.

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Table 2 Sample 0246 hours

Test tube 1 4.5-106 3.5-106 5.3-106 6.0-106

Test tube 2 4.5-106 3.5-105 5.3-104 5.6 * 10

As can be seen from the table, the number of bacteria in tube 2) after 6 hours is only 1/1000 of the number of test tube 1 / at the same time that no or only slight growth has occurred in tube 1), but instead bacteria have died in tube 2).

The effect of the antibacterial system was determined in a test tube. The various E. coli strains were then incubated at + 37 ° C in an aqueous solution of 5 g whey powder / 100 ml, 21 ppm sodium thiocyanate and sodium percarbonate in an amount corresponding to 0.25 mM H 9 ° 2 * E. coli. n mortality after 2 hours is given in Table 3 below.

Table 3 E. coli strain Percent death 0-139 299/66> 99.98 0-149 853/67> 99.98 0-138 355/67> 99.95 0-8 915/66> 99.99 0 -147 949/66> 99.99 0-141 220/65> 99.99

Several of the E-coli strains tested above are resistant to antibiotics. The results thus show a high degree of application of the system.

The high degree of novelization is also evidenced by the fact that the system also has an effect. > 40 mg LP / kg), 22.5% by weight of whey powder, 12.5% by weight of soya powder, 15.0% by weight of animal fat, 4.5% by weight of lactose and 0.5% by weight of vitamins. The strain was incubated at + 37 ° C in 10% aqueous milk replacer with 5 μg / ml lactoperoxidase. Two different concentrations of sodium thiocyanate and three different concentrations of H ~ 0o in the form of sodium percarbonate were used, as shown in Table 4 below.

Table 4

Concentration in mM Number of bacteria / ml SCM K2 ^ after 2 hours after 2 hours 0.25 0 2.5-106 7.7-107 0.25 0.25 2.5-106 5.3-104 0.50 0 1.8-106 1.0-108 0.50 0.50 1.8-106 1.5-104

As shown in Table 4, the antibacterial system gives a strong bactericidal effect at both concentrations tested, while the address vt u is that 3'IN does not give any effect per se, but bacterial growth continues when no peroxide-releasing substance is present. . ^ j * '

The peroxide donor may also be an alkime. The reaction which gives hydrogen peroxide sulphate in the reaction.

64272 For the first time, the transfer price for the first time in the middle of the year and in the host channel Akti -verbart antibakter.iel.lt system.

In particular, a particular barrel can be found in the gastrointestinal tract to be infected with E. coli and salmonella arteries. Hittills kända sätt att eliminera dessa infektioner är därvid att t.Lil-föra terapeutisk mängd antibiotika, med ätföljande risk för uppkomsten av antibiotikaresistenta bacterialammar.

In this case, the bacterial infectious genomic uptake of the bacterial infection system can be performed in such a way that the bacterial infection system can be used as an agent in the field of infection. , 1 mg / kg in rodents (50 ^ -5000 U / kg in rodents, thiocyanate in the presence of 16 g of sodium hydroxide and fast, water peroxide donor, in addition to alkali percarbonate, jordalkaliperoxide or 21 parts of urea peroxide in the game) beräknac som sodium sodium percarbonate.

Dot mol era förhällandet mellan peroxiddonator och tiocyanat är företrädesvis mindre än 4: 1, företrädesvis 1-2: 1-

Enrichments in the formulation of thiocyanates in the form of 160-3500, in the case of .160-1750 ppm / kg in the form of NaSCN, peroxide in the formulation of 210-4000, in the case of 210-2000 ppm / kg of the formulation perkarbonn.t och lactaktoxidas i en mängd av 10-200, förcträdos-vis 10-100 mg / kg.

When the thiocyanate is released, a digestion channel with a concentration of 0.1, 0.2-0.4 mM is used, and the concentration is converted to a toxic concentration (10 m.M.). When the peroxide donor is released, an equimolar solution of H 2 ° 2 is used, with a concentration of 0.1, 0.2-0.4 mM. The lactoperoxide game contains 8 64272 enzymes of activity, which can be based on the addition of 1 mg of enzyme to 50 enhancers (U), with 1 mg of enzyme per liter of digestion.

This solution has a thiocyanate concentration of 0.1, 0.2-0.4 mM for fatty acids, 100% peroxide donor concentration for 0.1, 0.2-0.4 mM for fatty acids, 50 mg of lactose peroxide concentration (50 U / l). For example, it can vary from 1 to 2 mg / l (50-100 U / l).

In the case of lactoper peroxides, lactoperoxide containing 1 mg of pyrogallin in pyrogallol under 80 seconds at a pH of 6.0 and 20 ° C is not required.

Mängden lactaktoxidas i vassle frän bovin mjölk (opastöri-serad) eller i ultratiltrerad vassle kan helt naturligt variera. The active ingredient in lactated oxide is enriched in about 30 mg / l, and then the lactoperoxide in milk is enriched in 1/30 to 1 mg.

The application of the exemption is limited to those exemptions.

Example

In addition to this, the weight of the above ingredients is 50.5 kg of whey powder 19.0 kg of skimmed-milk powder (z. 100 mg LP / kg) 14.0 kg of flaxseed 8.0 kg of lactose 7.5 kg of ready-to-use rice 1, Vitamin och mlneral 100.0 kg and ?] g of sodium sodium carbonate 16 g of sodium thiocyanate 9 64272

Ingredients in the field. For this purpose - bleaching in water at a temperature of 30-35 ° C with .10-15% water content (10-15% water content) and in the following areas: play of about 0,2 liters 4-5 gangers om dygnet.

Game sodium carbonate can vary from 210 to 420 ppm and sodium thiocyanate from 160 to 320 ppm, the colors vary by equimolar,

The exemplified sodium percarbonate is then reacted with the polymer genome to form a granule with a solid permeability to the polymer.

The polymer used is a wax, cellulose acetate, phthalate: a liquid and a permeable polymer.

Vidare kan dubbelskikt användas om en uppslamnlng fore admin-nistrering skulle ge en polymerlösande miljö.

Biologisk e £ fekt

For example, 10 ml of water containing 21 pom (0.25 mM) of sodium thiocyanate at 30 ° C, inoculated with Ps. Fluorescent EF 1998, is added. Sodium percarbonate is dissolved in 0.1, 0.2 or 0.3 mM H 2 O 2 and treated with water. The game is characterized by the inoculation as well as 2 inches after inoculation. Lactoperoxxas can be found in the nervous system at 5 μg / ml. The results are shown in Table 1 below.

/

Table 1

Sodium percarbonate in en Anthelmintic / ml game motsvarande min H2C> 2 0 Tim 2 Tim

Control (0.0 mM) 2.1 · 10 ^ 2.1 * 10 ^ 0.1 mM 2.1 · 106 1.8 · 105 0.2 mM 2.1 · 106 2.8 103 0.3 mM 2.1 * 106 2.6 - 1C3 10 64272

In this case, an in vitro antibody is used which has an anti-bacterial effect of 0.2 mM in non-bacterial nerves.

Reduction was performed at 0.1 mM with significant bactericidal effects.

Ps. Fluorescent EF 1998 incubated at + 30 ° C in the presence of 1) and 2), colors 1) in the form of a commercially available product containing 15% ultrafiltrated copper powder, 57% copper powder, 8% fat , 16% animal protein, 4% vitamins and mlaler in 10 ml of water, colors with 50 ml of lactoperoxide velvet with sodium sodium carbonate 27.5 ppm and colors with 2) ), 21 parts of sodium thiocyanate and 27.5 ppm of sodium percarbonate.

The bacteria in the sample are replaced by a table of 2 to 2 results, 0, 2, 4 and 6 in the table,

Table 2

Prov 0246 timmar provrör 1 4,5 · 106 3,5. 106 5.3 * 106 6.0 · 106 Provrör 2 4.5-106 3.5-105 5.3-104 5.6 * 103

In the case of a table which is bacterial in the body 2) after the test is 1/1000 of the test in the body 1) at the same time, the color is shown in the body 1).

The antibacterial system has an effect in vitro. When incubated, E. coli was incubated at + 37 ° C in a water-free solution of 5 g of copper powder / 100 ml, 21 ppm of sodium thiocyanate and sodium impercarbonate and 0.25 mM ^ 2 ^ 2 '^ esu · * · "in the case of E. coli as specified in Table 2.

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Table 3 E. coli strain Percentage from 0-139 299/66> 99.98 0-149 853/67> 99.98 0-138 355/67> 99.95 0-8 915/66> 99.99 0 -147 949/66> 99.99 0-141 220/65> 99.99

Flora of the test E. coli strain is resistant to antibiotics. The results can be used to determine the value of the system.

Hög användningsgrad föreligger även genom att systemet utövar effekt även pä gramposltiva bacteracteria.

A test of E. coli 9703 was performed on a caloric acid-free product containing 45% w / w skummjölkspxilver (> 40 mg LP / kg), 22.5 wt.% Vasslepulver, 12.5 wt.% Soy powder , 15,0% by weight of animal fat, 4,5% by weight of lactose and 0,5% by weight of vitamins. Incubate at + 37 ° C for 10 minutes with a volume of 5 μg / ml. 1 actoperoxide. ^ vs olika concentrate av natriurntiocyanat ooh tre oi age Concentrator av i sodium percarbonate tower användes, säsom av nedanstaende tabell 4 framgär.

Table 4

Kone e n hr at i o n j jmM Antal bakteri.er/ml SCI ·: H „0 ,. efter 0 Tim efter 2 Tim z 2 0.25 0 2.5 · 106 7.7 * 107 0.25 0.25 2.5 * 106 5.3 * 104 0.50 0 1.8 · 106 1.0 * 108 0.50 0.50 1.8 · 106 1.5 * 104 12 64272

In Table 4, an antibacterial system and a kraft bactericidal effect are used to test the concentration of the same mixture at the SCN and in the case of large grains. the effect of the bacterial dietary action of the peroxide donor is on the nerve pathways,

For example, oxide donors can be treated with alkali metal sulfate to react with sulfur peroxide and sulfate.

Claims (9)

A process for the preparation of nutrients containing an antibacterial activatable in the gastrointestinal tract. system, characterized in that nutrient compositions known per se are added to an antibacterial system containing lactoperoxidase in a range of dietary mino 1 mg / kg nutrient (50-5000 U / kg nutrient), a thiazyanate in an amount of at least 16 ppm. calculated as pure methyl thiocyanate and a solid, water-soluble peroxide donor, which are alkali percarbonate, soil or peroxide or urea peroxide, in an amount of at least 21 ppm calculated as sodium percarbonate. Method according to claim 1, characterized in that lactoperoxidase is used in pure form. 3. A method according to claim 1, characterized in that lactoperoxidase is used in the form of a 1 ac. operoxidase containing milk product.