GB1576528A - Vitamin product - Google Patents

Description

(54) VITAMIN PRODUCT (71) We, FARMOS-YHTYMA OY of P.O. Box 425, SF-20101 Turku 10, Finland, a joint stock company organized under the laws of Finland, do hereby declare the invention, for which we pray that a patent may be granted to us, and the method by which it is to be performed, to be particularly described in and by the following statement: The present invention concerns a stabilized vitamin product especially useful in the feeding of animals. The product is extremely advantageous as regards its stability, and its absorption, accumulation and utilization in the animal body and also its palatability when containing e.g. vitamins A, D, E and K.

The use of vitamin solutions which contain fat soluble vitamins and which are made "water soluble" through emulsifying has increased in the feeding of animals. This is due i.e. to the automatization of feeding which is a result of increased animal stock units. The su plemental vitamin requirement has increased as a result of increased production levels (stress) and improved fodder utilization. Supplemental vitamin feeding has become routine and it is in most cases easiest to administer vitamins separately as a liquid fodder preparation together with the drinking water.

Orally administrable emulsions containing vitamins A, D, E (and sometimes K) or mixtures thereof are conventionally prepared by incorporating an oil vitamin raw material together with an emulsifier while vigorously stirring them into a base or vehicle, usually water. When comparing the resulting "water soluble" products with respect to their utilization, usually only the absorption capacity of the emulsified vitamin (for example vitamin A acetate/palmitate) has been considered, but not the possible effect of the base.

Surprisingly it has now been found in accordance with the invention that by incorporating into the base of the vitamin emulsion one or more sugar alcohols including xylitol it is possible to improve the stability i.e. shelf life, the resistance to microbial attack, the palatability and most importantly, the utilization by the animal body of the vitamins.

Preferably the emulsion contains at least 2.25% by weight based on total weight of the emulsion. There may be e.g. from 5% to 15% or more of xylitol.

The total sugar alcohol content is preferably from 5 to 30% by weight of toatal weight e.g.

15%. It has proven especially advantageous to add xylitol as well as or as an ingredient of a sugar alcohol mixture obtained as a by-product in the xylitol production from plant material such as birch wood, into the emulsion base. Further preferred features of the invention appear from the appended claims.

The above mentioned by-product can be obtained in the form of a yellowish brown, sweet liquid having a specific weight of about 1.22 as a 50% aqueous solution. Its caloric value and degree of sweetness are the same as for glucose. The by-product contains several sugar alcohols and usually its composition, calculated as dry matter, is the following: Table I xylitol 15-25% by weight arabitol 20-35% by weight mannitol 15-25% by weight sorbitol 5-15% by weight dulcitol 5-15% by weight rhamnitol 5-10% by weight others 2-5% by weight decomposition products 2-5% by weight Thus emulsions containing 15 % of this by-product, as set out in the Examples which follow, will contain from 2.25 to 3.75% by weight of total weight, of xylitol, and may contain up to 7.5% if-the emulsion contains 30% of this by-product.

In more recent processes the recovery of xylitol has been improved and the composition, calculated as dry-matter, may vary within the following lines: Table II xylitol 6-18%byweight arabitol 9-21% by weight mannitol 13-19% by weight sorbitol 8-12% by weight dulcitol 5-11% by weight rhamnitol 4-6 % by weight reducing sugars 8-15%byweight other polyols 6-14% by weight From literature it is known e.g. that xylitol stimulates the functions of the liver and enhances cell activity. It is also known that microbes in the mouth are not able to utilize xylitol as an energy source and that in rumen fluid (in vitro) sugar alcohols are preserved without decomposing for long time periods (Poutiainen E., et. al., 1976, Proc. Nutr. Soc. 294th Scientific Meeting, London, p. 140-141 A and SF-patent application 76 0746, U.K. Patent Specification 1,542,802).

The invention is illustrated by means of the following Examples concerning additives which have proves to be especially advantageous.

Example 1 - Resistance to microbial growth.

In Example 1 microbial growth in aqueous solutions of different base additives of the vitamin emulsions was investigated. Sample solutions containing xylitol alone and as an ingredient of the above described by-product from the xylitol production were compared with samples containing saccarose and containing only water.

In these tests a solution containing 15%by weight of xylitol, of by-product or of saccarose as dry matter as well as a sample containing distilled water, were contaminated with a microbe suspension containing the following microbes: Escherichia coli (gram-negative rod) Staphylococcus epidemis figram-positive coccus) Saccharomyces carlsbergensis (yeast).

One night old pre-cultures were stirred to form a mixed suspension which was diluted in the ratio of 1 to 1000. 1 ml of this suspension was pipetted into 100 ml of each test solution and the solutions were transferred to an incubation chamber, the temperature of which was 35"C.

The microbe concentrations in the solutions were followed for over one month by carrying out base cultivations on bacteria(count-agar) and on yeast/mould (saboraud-agar). The results of the tests are shown in Table III. In the table the figures show the microbe concentration at each time determined as number of cells/ml.

Table III Microbial growth in different vitamin emulsion base additives

Incu- Base additive bation time, by-product distilled Xylitol Saccarose days water Cells/ml Cells/ml Cells/ml Cells/ml bact- yeast/ bact- yeast/ bact- yeast/ bact- yeast/ eria mould eria mould eria mould eria mould 1 21 000 500 25 000 3 000 22 000 600 35 000 200 2 100 300 500 100 300 200 4 300 3 700 9 60 1260000 170 110 90 60 6 000 000 100 000 i4 1 340 000 180 120 120 200 54000 34000 41 - - - - - - - - From the test results it may be concluded that when comparing saccarose, xylitol, the by-product from the xylitol production and distilled water, the saccarose solution proved to be clearly the best growth substate for the microbes. Xylitol and distilled water were mutually equivalent, which proves that xylitol is a bad microbe nutrient, but under the test conditions the xylitol does not seem to inhibit the growth of the microbes to any larger extent either. The by-product obtained from the xylitol production apparently contains components inhibiting the growth of bacteria but not of yeasts. It may thus be summarized that the bacteria thrive badly on sugar alcohols, that is at least as badly as in pure water.

Example 2 - Resistance to microbial growth In this test two vitamin solutions were compared as regards their tendency to become contaminated. The preparations used were a water-based vitamin emulsion (A-D-E-VIMIN (registered trade mark) Astra) and a corresponding vitamin A, D, E emulsion containing 15% of sugar alcohols (by-product from the xylitol production, Table I) as dry matter calculated on the total weight of the emulsion. The contamination and experimental methods were the same as in Example 1. The results are shown in Table IV.

Table IV

Incub- Vitamin emulsion tion time, A-E-VIMJN /Astra A-D-F-emulsion, containing days by-product Cells/ml Cells/ml bacteria | yeast/mould bacteria yeast/mould 1 52 000 000 25 600 000 31 000 700 2 750 000 40 000 110 100 9 190 000 160 000 14 240 000 110000 - 41 - 82 000 - From the results it may be seen that water based A-D-E VIMIN is a much better growth substrate for the microbes than the corresponding sugar alchol containing vitamin A, D, E solution. From the sugar alcohol containing vitamin emulsion the microbe growth disappeared very quickly after contamination. After one day the bacteria content had decreased to less than 1/100 of the amount after contamination, that is, the sugar alcohol containing vitamin emulsion is microbiologically very stable.

The same kind of contamination test was also carried out using a periodical contamination procedure with two additional known reference vitamin products. The test results are shown in Table V.

Table V Growth of microbes in different vitamin products Time Vitamin product days

A-D-vitamin A-D-E-vitamin A-D-vitamin A-D-E-vitamin Orion Astra Rohto 15 % sugar alcohols Mixed suspension Mixed suspension Mixed suspension Mixed suspension Count-agar Saboraud- Count-agar Saboraud- Count-agar Saboraud- Count-agar Saboraudbacteria agar bacteria agar bacteria agar bacteria agar cells/ml yeast/mould cells/ml yeast/mould cells/ml yeast/mould cells/ml yeast/mould cells/ml cells/ml cells/ml cells/ml 1 < 100 < 100 900 90 000 5 000 40 000 < 100 < 100 2 (conta- 600 600 90 000 2 000 000 800 2 000 < 100 < 100 mination 3 150 550 20 000 2 000 000 < 100 5 000 < 100 < 100 9 < 100 < 100 200 000 200 000 < 100 < 100 < 100 < 100 10 (cont.) 3 000 2 000 250 000 250 000 2 000 2 000 2 000 2 000 11 3 500 2 000 100 000 1 000 < 100 < 100 < 100 < 100 17 < 100 < 100 100 000 300 000 < 100 < 100 < 100 < 100 18 (cont.) 3 000 000 700 000 2 000 000 600 000 2 000 000 800 000 200 000 70 000 19 1 500 000 25 000 1 000 000 250 000 < 100 300 < 100 < 100 24 - 500 000 40 000 < 100 < 100 < 100 < 100 25 (cont.) 3 000 000 900 000 1 000 000 4 000 000 4 000 000 700 000 400 000 90 000 26 4 000 000 600 000 2 000 000 1 000 000 5 000 4 000 < 100 < 100 32 200 000 20 000 800 000 500 000 < 100 < 100 < 100 < 100 49 < 100 < 100 400 000 2 000 000 < 100 < 100 < 100 < 100 The addition of sugar alcohol makes the vitamin product very stable against microbes. The contamination disappeared each time in one day.

Example 3 - The stability of vitamins In this test the preservation of vitamins was investigated at different temperatures when sugar alcohol (by-product of the xylitol production, Table I) was added to the vitamin emulsion base in an amount of 15%by weight of dry matter, based upon the total weight of the emulsion. As vitamin components the emulsion contained 30 000 IU of vitamin A, 3000 IU of vitamin D3 and 30 mg of vitamin E per ml. Distilled water was used as the base into which the sugar alcohol and the other necessary agents were added.

The preparation was frozen (-20 C) and kept at this temperature for 10 days, whereafter the preparation was thawed and kept at room temperature (+20 C) for 53 days, whereafter the product was kept in a refrigerator (+4 C). Samples were taken from the preparation during the freezing period (3 days), after storage at room temperature (53 days) and after storage in the refrigerator whereby the product had been stored for a total period of 9 months. The samples were tested as to their vitamin content at each stage. The results are shown in the following table.

Table Vl The preservation of a vitamin emulsion at different temperatures

Storage time Temperature Vitamin E Vitamin A C 3 days -20 36.7 32 900 53 days +20 34.7 31 600 . 9 months +4 33.9 31 460 From the test results it may be concluded that the vitamin content of the preparation is preserved for extended times at varying temperature conditions.

Example 4 - Movement of radioactivity labelled vitamin A in the alimentary tract In these tests the movement of radioactivity in the alimentary tract was investigated in A, D, E preparations by means of tritium-labelled vitamin A, Al (All-trans) 1-3H(N), using three groups each of five SPF (specific pathogen free) mice. The results of the test are given in the fqllowing Table VII which shows the radioactivity found in the alimentary tract two hours after peroral administration, calculated as weight Wo of the administered amount. As the base additive of the vitamin emulsion, xylitol and the by-product from xylitol production (Table I) were used, and also a test series as performed in which pure water was used as the base. The sugar alcohol in each case constituted 15 No as dry matter of the total weight of the emulsion.

Table VII The movement of labelled vitamin A in the alimentary tract

Part of the Radioactivity alimentary tract Xylitol By-product Pure Water stomach 3.40i0.24X 4.64i0.l2XX 5.88+0.20 small intestine, 6.50i0.26 6.26+0.24 7.56+1.00 front part large intestine, 2.86+0.08 3.12+0.16 2.62i0.16 back part large intestine 2.76i0.02 3.16+0.12 3.68+0.38 and rectum total content 15.52 17.16 19.74 The values of the Table are mean values + SEM (Standard Error of Mean).

The xylitol and by-product levels in the stomach differ statistically significantly from the values obtained with pure water. The significance of difference between xylitol and water is (Student'st-test) P < 0.05 and the corresponding value for the by-product is P < 0.01, that is the different is statistically very significant. The values for the other parts of the alimentary tract do not differ significantly from the values obtained with water. Since significantly less radioactivity was found in the stomach of the test animals when the base of the marked vitamin emulsion contained sugar alcohol, it follows that the vitamin has disappeared either through absorption into the organism or with the excrement. Tests on the excrement follow.

Example 5 - The secretion of radioactivity labelled vitamin A with excrement This test followed according to Example 4 the transition if isotopically labelled vitamin A to the excrement of the test animals. A vitamin preparation was administered according to Example 4 perorally to groups of five SPF-mice and the excrement samples were collected daily for 30 days. The radioactivity results of the collectively obtained excrement samples are shown in the following Table VIII.

Table VIII The radioactivity of excrement samples of the test animals Time, day Xylitol By-product Pure Water 1st 7.62 8.86 13.20 2nd 0.04 0.06 0.12 3rd - Although the radioactivity in the case of the emulsions according to the invention the bases of which contained xylitol and the by-product from the xylitol production had thus disappeared from the intestines (absorption area) faster than that of a pure water based emulsion, as shown by Example 4, a corresponding increase was not found in the excrement, rather the opposite, as shown by Table VIII.

In the urine there was secreted less than 1% of the amount administered. There was no statistically significant differences between the groups.

Example 6 - The absorption of tritium labelled vitamin A into the organism From the preceeding Examples it could be expected that the labelled vitamin would move faster into the blood serum, liver and other inner organs from the xylitol and by-product based emulsions than from water based solutions.

In order to establish this, SPF-mice were administered perorally with single doses of 450 IU of vitamin A (tritium labelled) as a vitamin A, D, E solution and the radioactivity of the blood serum as well as that of the liver was determined as weight % of the dose, 2 and 6 hours as well as-1, 2, 3 and 7 days after administration. The results are shown in Table IX and in Figure 1.

Table IX The labelled vitamin A in blood serum Time Xylitol By-product Pure Water 2 hours 0.38 i 0.02Xxx 0.29 + 0.01 0.29 + 0.02 6 hours 0.30 i 0.03 0.28 i 0.01 0.26 j 0.01 .1 day 0.16 f 0.01 0.16 < i 0.01 0.16 + 0.01 2days 0.12 j 0.01 0.13d+ 0.01 0.13 + 0.01 3 days 0.08 < i 0.01 0.09 < i 0.01 0.08 < i 0.01 7 days 0.06 < i 0.01 0.05 < * 0.01 0.07 j 0.01 In the Table the value for xylitol after 2 hours differed statistically to a very significant degree (P 00001) from the corresponding value for water. The corresponding value for the by-product is inconsistent. As to the other values, no significant differences could be found.

In Figure 1 the corresponding radioactivity results for the liver are shown graphically. In the Figure the results are mean values + SEM. The results obtained with xylitol after 2 and 6 hours differ statistically to a significant degree from the results obtained with pure water.

The amounts of radioactivity detected in the kidneys, the brain and the heart remained less than 1%. There were no statistically significant differences between the groups.

From Examples 4 to 6 it may be concluded that sugar alcohols, especially xylitol as well as the by-product formed during the production of xylitol from plant material, stimulate the absorption of vitamins from the intestines, as well as their accumulation into the organism and their utilization therein.

The results indicate that so far as the physiological up-take is concerned, the actual content of xylitol is critical to succes.s In particular the figure showing up-take in the liver shows that when the pure yxlitol is diluted with other sugar alcohols and reduced in its overall proportion, the up-take is substantially reduced, although it remains at a higher level than where it is absent. The results for secretion are also demonstrative of the criticality of the presence of xylitol. In the latter experiments the benefit is largely obtained even with the reduced amount (2.25 to 3.75%) of xylitol in the polyol, showing that even at this xylitol level, significant improvements in utilization of the vitamins overall, is achieved, whereas to ensure utilization by the liver, a ligher proportion of xylitol gives added benefit.

Claims (13)

WHAT WE CLAIM IS:

1. A vitamin product in the form of an aqueous emulsion containing one or more vitamins and one or more sugar alcohols including xylitol.

2. Avitamin product according to claim 1 containing from 2.25 to 30% by weight of total weight of xylitol.

3. A vitamin product according to claim 1 containing at least 5 by weight of total weight of xylitol.

4. A vitamin product according to claim 1 containing at least 15% by weight of total weight of xylitol.

5. A vitamin product according to any preceding claim wherein the total content of sugar alcohols is from 5 to 30% by weight of total weight.

6. A vitamin product according to any preceding claim containing xylitol, arabitol, mannitol, sorbitol, dulcitol and rhamnitol.

7. A vitamin product according to claim 5 wherein at least a portion of the xylitol present derives from a mixture of sugar alchols obtained from the commercial production of xylitol from plant material and comprising: xylitol 15 - 25 % by weight arabitol 20 - 35% by weight mannitol 15 - 25% by weight sorbitol 5 - 15% by weight dulcitol 5 - 15% by weight rhamnitol 5 - 10% by weight

8. A vitamin product according to claim 5 wherein at least a portion of the xylitol present derives from a mixture of sugar alcohols obtained from the commercial production of xylitol from plant material and comprising: xylitol 6 - 18% by weight arabitol 9 - 21% by weight mannitol 13 - 19% by weight sorbitol 8 - 12% by weight dulcitol 5 - 11% by weight rhamnitol 4 - 6% by weight

9. A vitamin product according to claim 7 wherein the amount of the mixture incorporated in the product is substantially 15% by weight of total weight.

10. A vitamin product according to any preceding claim containing vitamins A, D and E.

11. A vitamin product according to claim 10 containing vitamin K.

12. A vitamin product according to claim 1 substantially as described herein with reference to any one of the Examples.

13. A foodstuff for animals containing a vitamin product according to any preceding claim.