OILY FORMULATION COMPRISING AT LEAST ONE PROBIOTIC BACTERIUM

Abstract

An oily formulation comprising at least one probiotic bacterium and comprising a first oily phase comprising at least one saturated or hydrogenated vegetable oil in which is incorporated said at least one probiotic bacterium, a second oily phase comprising at least one polyunsaturated vegetable oil different from said saturated or hydrogenated vegetable oil, and at least one dispersant.

Claims

1. An oily formulation, comprising: a first oily phase comprising at least one vegetable oil in which is incorporated at least one probiotic bacterium, wherein the at least one vegetable oil is selected from the group consisting of a saturated vegetable oil and a hydrogenated vegetable oil; a second oily phase comprising at least one polyunsaturated vegetable oil different from said at least one vegetable oil, and; at least one dispersant; and said formulation being characterized in that said first oily phase is formed by a mixture of a hydrogenated palm oil and of a hydrogenated coconut oil, said mixture having a melting temperature between 20° C. and 30° C. and being present in an amount ranging from 2% to 5% by weight based on a total weight of said formulation.

2. The oily formulation according to claim 1, characterized in that the oily formulation has a dynamic viscosity between 0.1 P.Math..s and 0.35 Pa.Math.s.

3. The oily formulation according to claim 1, characterized in that said at least one dispersant is a lipophilic non-ionic surfactant selected from the group consisting of polysorbates, lecithin, sorbitans, sorbitan esters and mixtures thereof.

4. The oily formulation according to claim 1, characterized in that said at least one dispersant is present in an amount ranging from 0.5 to 2% by weight based on the total weight of said formulation.

5. The oily formulation according to claim 1, characterized in that said at least one polyunsaturated vegetable oil is selected from the group consisting of soybean oil, canola oil, sunflower oil, macadamia oil, groundnut oil, grape pip oil, pumpkin seed oil, flax oil, olive oil, corn oil, safflower oil, sesame oil, pine kernel oil, conjugate linoleic acid, evening primrose oil, almond oil, peach kernel oil, apricot kernel oil, nut oil, rapeseed oil, raspberry seed oil, blueberry seed oil, cranberry seed oil and other fruit seed oils, fractionated coconut oil and mixtures thereof.

6. The oily formulation according to claim 1, characterized in that said at least one polyunsaturated vegetable oil is present in an amount of more than 90% by weight based on the total weight of said formulation.

7. The oily formulation according to claim 1, characterized in that said at least one probiotic bacterium is a probiotic bacterium selected from the group consisting of a microencapsulated probiotic bacterium and a coated probiotic bacterium.

8. The oily formulation according to claim 1, characterized in that said at least one probiotic bacterium has a particle size of less than 600 μm.

9. The oily formulation according to claim 1, characterized in that said at least one probiotic bacterium is present in said formulation in an amount of 106 to 1010 CFU/ml of formulation.

10. The oily formulation according to claim 1, characterized in that it further comprises an additive selected from the group consisting of antioxidants, stabilizers, thickeners, nutrients, fatty acids and mixtures thereof.

11. A method for manufacturing an oily formulation comprising at least one probiotic bacterium, said method comprising the steps: preparing a pre-mix formulation by mixing and incorporating at least one probiotic bacterium into a first oily phase formed with a mixture of hydrogenated palm oil and hydrogenated coconut oil, said mixture having a melting temperature between 20° C. and 30° C. and being present in an amount ranging from 2% to 5% by weight based on a total weight of said pre-mix formulation, and mixing said pre-mix formulation into a second oily phase comprising at least one polyunsaturated vegetable oil with addition of at least one dispersant.

12. The manufacturing method according to claim 11, characterized in that it comprises a step of preparing said at least one probiotic bacterium prior to said step preparing the pre-mix formulation that is selected from the group consisting of a microencapsulation step and a coating step.

13. The manufacturing method according to claim 12, characterized in that said coating step is carried out by applying a coating selected from the group consisting of alginate, chitosan, pectin, pullulan, gelatin, carrageenan, agar-agar, cellulose, hemicellulose, ethylcellulose, carboxycellulose, and mixtures thereof.

14. The manufacturing method according to claim 11, further comprising: conditioning said oily formulation by encapsulating in a soft gelatin capsule.

15. The manufacturing method according to claim 14. characterized in that the oily formulation has a dynamic viscosity between 0.1 Pa.Math.s and 0.35 Pa.Math.s.

16. The oily formulation according to claim 2, characterized in that the oily formulation has a dynamic viscosity of 0.18 Pa.Math.s.

17. The oily formulation according to claim 2, characterized in that the oily formulation has a dynamic viscosity 0.15 Pa.Math.s at a temperature of 15° C.

18. The manufacturing method according to claim 13, characterized in that applying step comprises spraying the coating.

19. The manufacturing method according to claim 11, further comprising: conditioning said oily formulation by filling in an ampoule, or by filling a hard gelatin capsule, the core of which is liquid.

20. The manufacturing method according to claim 14, characterized in that said at least one dispersant is a lipophilic non-ionic surfactant selected from the group consisting of polysorbates, lecithin, sorbitans, sorbitan esters and mixtures thereof.

Description

EXAMPLES

1. Formulation According to the Invention

[0050] Table 1 below mentions the amounts of the different compounds for 100 ml of an oily formulation according to the invention:

TABLE-US-00001 TABLE 1 Compound Amount Function Sunflower oil 94 ml Second oily phase Saturated or hydrogenated 2.5 ml First oily phase coconut-palm oil (40/60) Lactobacillus rhamnosus 1 g Probiotic bacterium Sorbitan monolaurate 1 g A lipophilic non-ionic surfactant (dispersant) Alpha-tocopherol acetate 0.2 g Antioxidant Lycopene 0.04 g Antioxidant and coloring agent Cholecalciferol 0.001 g Vitamin D3 source

[0051] The probiotic bacteria Lactobacillus rhamnosus are incorporated into the first oily phase consisting in a 40/60 mixture of hydrogenated coconut oil and of hydrogenated palm oil in order to form a pre-mix for which the melting temperature is comprised between 20° C. and 30° C., which is then dispersed, by means of the presence of sorbitan monolaurate (dispersant), in a second oily phase consisting in a polyunsaturated oil which is sunflower oil.

[0052] Additives of the antioxidants and/or coloring agent type (alpha-tocopherol acetate, lycopene) and a D3 vitamin source are also added according to this embodiment of a formulation according to the invention.

2. Viability of the Probiotic Bacteria in a Formulation According to the Invention

[0053] Tests were conducted in order to measure over time the viability of probiotic bacteria for a formulation as described in example 1 and kept at two different temperatures (4° C. and 25° C.), this formulation never being subject to any stirring except just before the taking of the analyzed sample.

[0054] Three repetitions were carried out for each measurement of the viability of the probiotic bacteria (CFU/g). These viability measurements of the probiotic bacteria were carried out after 7, 14, 20, 27 and 63 days following the preparation of the formulation according to the invention. FIG. 1 resumes the obtained results.

[0055] As this may be seen, without any stirring, the viability of probiotic bacteria present in a formulation according to the invention is particularly high. Indeed, when the oily formulation is kept at a temperature of 4° C., a survival rate of 73% and of 58% are respectively reported after 27 and 63 days.

3. Comparative Stability Tests of Compositions According to the Invention Comprising Variable Amounts (% By Weight) of the First Oily Phase (Saturated or Hydrogenated Vegetable Oil)

[0056] Comparative tests were carried out by varying exclusively for a same basic formulation, the amount by weight of the first oily phase. Amounts by weight (% by weight) of 2.5%, 6% and 8% based on the total weight of the formulation were tested in order to determine whether the obtained formulation remains in a stable liquid form, i.e. if the obtained formulation does not solidify in which case the properties and the viability of the probiotic bacteria would be altered.

[0057] These tests were conducted according to three different temperatures, i.e. by exposing the formulations formed at 4° C., at 15° C. and at 25° C. for 24 h. The obtained results are resumed in Table 3 below while Table 2 resumes the different tested formulations.

TABLE-US-00002 TABLE 2 % by weight Formulation Formulation 1 Formulation 2 3 Sunflower oil 92.47 88.97 86.97 Mixture of hydrogenated 2.5 6 8 coconut oil and of hydrogenated palm oil, the mixture having a melting temperature comprised between 20° C. and 30° C. Beeswax 2.5 2.5 2.5 Lactobacillus rhamnosus 1.48 1.48 1.48 Sorbitan monolaurate 0.85 0.85 0.85 Alpha-tocopherol acetate 0.2 0.2 0.2

TABLE-US-00003 TABLE 3 Formulation 1 Formulation 2 Formulation 3  4° C. liquid phase solid phase solid phase 15° C. liquid phase solid phase solid phase 25° C. liquid phase semi-solid phase solid phase

[0058] As this may be seen, after 24 h, only the formulation 1 according to the invention comprising the first oily phase is an amount of 2.5% by weight based on the total weight of the formulation is maintained in a liquid and stable form (no precipitation, no aggregation, no sedimentation of probiotic bacteria) at temperatures of 4° C., 15° C. and 25° C. On the other hand, it was observed that the formulations 2 and 3 comprising 6 and 8% by weight respectively of a first oily phase based on the total weight of the formulation appear in a semi-solid or solid form at these same temperatures after 24 h.

4. The Formulation According to the Invention: Tests with Different Second Oily Phases (Polyunsaturated Vegetable Oils)

[0059] Several formulations were tested in order to determine whether, for a same first oily phase formed with a mixture of hydrogenated palm oil and hydrogenated coconut oil (the mixture having a melting temperature comprised between 20° C. and 30° C.), diverse second oily phases comprising a polyunsaturated vegetable oil may be suitable. It was determined, for each formulation, whether it remains in a stable liquid form (no precipitation, no aggregation, no sedimentation of probiotic bacteria) at 4° C., 15° C. and 25° C. after 24 hours.

[0060] The obtained results are resumed in Table 5 while Table 4 resumes the different tested formulations.

[0061] As this may be seen, regardless of the polyunsaturated vegetable oil forming the second oily phase, stable liquid formulations (no precipitation, no aggregation, no sedimentation of probiotic bacteria) are systematically observed after 24 h for the temperatures of 15° C. and 25° C. Let us note that at the temperature of 4° C., the formulations 5 and 8 appear in a semi-liquid form. Moreover, it emerges from this example that regardless of the dispersant used, stable liquid formulations are systematically obtained at each of the tested temperatures.

TABLE-US-00004 TABLE 4 % by weight Formulation 4 Formulation 5 Formulation 6 Formulation 7 Formulation 8 Second oily phase Sunflower Olive oil Mixture of Groundnut Soybean oil oil 91.32 oils* oil 91.32 92.47 92.47 91.32 First oily phase: mixture 2.5 2.5 2.5 2.5 2.5 of hydrogenated coconut oil and hydrogenated palm oil, the mixture having a melting temperature comprised between 20° C. and 30° C. Beeswax 2.5 2.5 2.5 2.5 2.5 Lactobacillus rhamnosus 1.48 1.48 1.48 1.48 1.48 Dispersant Sorbitan Lecithin Sorbitan Lecithin Lecithin monolaurate 2 monolaurate 2 2 0.85 0.85 Alpha-tocopherol acetate 0.2 0.2 0.2 0.2 0.2 *MCT (Medium Chain Triglyceride)

TABLE-US-00005 TABLE 5 Formulation 4 Formulation 5 Formulation 6 Formulation 7 Formulation 8  4° C. Liquid Semi-liquid Liquid Liquid Semi-liquid phase phase phase phase phase 15° C. Liquid Liquid Liquid Liquid Liquid phase phase phase phase phase 25° C. Liquid Liquid Liquid Liquid Liquid phase phase phase phase phase

[0062] It is quite understood that the present invention is by no means limited to the embodiments described above and that many modifications may be made thereto without departing from the scope of the appended claims.