Claims:We Claim:
1. A nutritional suspension composition comprising:
I. At least one live probiotic bacterium,
II. At least one low or zero calorie sweetener,
III. At least one flavor,
IV. At least liquid dispersion medium,
Wherein, the said composition is packaged in a sterile blow fill seal tube, transparent or colored for market differentiation.
2. The composition according to claim 1, wherein said at least one low or zero calorie sweetener is selected from the list consisting of sucralose, saccharin, neotame, cyclamate, advantam, aspartame, acesulfame potassium (Ace K), alitame, steviol glycosides and thaumatin.
3. The composition according to claim 1 or 2, wherein said at least one low or zero calorie sweetener is sucralose.
4. The composition according to any preceding claims, wherein said composition comprises 0.01% to 10% w/w of said at least one low or zero calorie sweeteners.
5. The composition according to claim 1, wherein said at least one flavor is selected from the list consisting of cherry, maple, orange, pineapple, raspberry, banana, vanilla, chocolate, butterscotch, coconut, custard, cinnamon, strawberry, anish, apricot, honey, lime, coriander, lemon and combinations thereof.
6. The composition according to claim 1, wherein said at least one flavor is orange.
7. The composition according to any one of the preceding claims, wherein said at least one probiotic bacterium is Bacillus clausii.
8. The composition according to any of the preceding claims, wherein one dose of said composition comprises 10e3 to 10e12 colony forming units of said microorganism, such as 10e6 to 10e12 colony forming units, for example 10e7 to 10e10 colony forming units.
9. The composition according to any of the preceding claims, wherein said composition
I. 1% to 5% w/w of a mixture of Bacillus clausii and Bacillus sporogenesis.
II. 0.01% to 10% w/w sucralose.
III. 1% to 3% w/w orange flavors.
IV. Rest is purified water.
10. A method for preparing a suspension composition containing a probiotic bacterium, said method comprising the steps of
I. Aseptic mixing at least one microbial organism with at least one low or zero calorie sweeteners, at least one liquid dispersion medium and optionally one or more flavors, optionally one or more aroma compound, optionally one or more antioxidant agent.
II. Filling the suspension in a compatible container maintaining the aseptic condition and sealing.
, Description:Flavored Probiotic compositions
Field of Invention
The present invention relates to the field of nutrition. In particular, the present invention relates to the field of oral nutritional supplemental related to improved probiotic compositions.
Background of the Invention
Probiotics are live microorganisms or microbial mixtures administered to improve the patient’s microbial balance, particularly the environment of the gastrointestinal tract. The presence of ex. Bacillus is important for the maintenance of the intestine microbial ecosystem. Bacilli have been shown to possess inhibitory activity towards the growth of pathogenic bacteria such as Listeria monocytogenes, Escherichia coli, Salmonella spp and others. This inhibition could be due to the production of inhibitory compounds such as organic acids, hydrogen peroxide, bacteriocines or reuterin or competitive adhesion to the epithelium.
A variety of compositions for supplementing probiotics are currently available. The compositions are typically provided for improving the microflora within a gastrointestinal (Gl) tract of subjects. Current formulation technologies include the utilization of encapsulation and stabilization techniques for shielding the probiotics with a protective layer such that the composition comprising the microorganism may be delivered to Gl tract of the subject. Further, the focus of many formulation technologies has been to protect the viability of probiotics during distribution and storage.
However, there is a remaining need for formulation of probiotics having properties that will associate the ingestion with a pleasant taste and mouth feel.
Summary of the Invention
The present invention was made in view of the prior art described above, and the object of the present invention is to provide a probiotic formulation having improved taste and mouth feel.
To solve the problem, the present invention is a nutritional composition as described in the below first aspect.
A first aspect of the present invention concerns a nutritional composition comprising:
I. At least one live microbial organism,
II. At least one zero or low calorie sweetener,
III. At least one flavor,
IV. At least one liquid dispersion medium.
The composition may optionally comprise one or more aroma compound and/ or one or more acidity regulator and/ or one or more antioxidant agent.
A second aspect of the present invention concerns a container containing the composition of the present invention.
A third aspect of the present invention provides a method for preparing a composition containing a microbial organism, said method comprising the steps of
I. Aseptic mixing of at least one microbial organism with at least one low or zero calorie sweetener, at least one flavor, at least one liquid dispersion media and optionally one or more aroma compound and/ or one or more acidity regulator and/ or one or more antioxidant agent.
II. Filling suspension in a compatible container maintaining the aseptic condition sealing.
The composition obtainable from the above method is provided in a further aspect.
Yet further aspects of present invention concerns method for and use of the composition of the present invention for preventing and treating various gastrointestinal disorders as an oral bacteriotherapy.
Detailed Description of the Invention
In describing the embodiments of the invention specific terminology will be resorted to for the sake of clarity. However, the invention is not intended to be limited to the specific terms so selected, and it is understood that each specific term includes all technical equivalents which operate in a similar manner to accomplish a similar purpose.
Definitions
Microbial Organism
In the context of the present invention the term “Live microbial organism” refers to a microorganism that when ingested in adequate amounts by a subject (such as in the form a formulation as described herein) confer a health benefit to the subject. A probiotic microorganism is a live microorganism which, when administered in adequate amounts, confer a health benefit to the host by influencing the composition and or metabolic activity of the flora of the gastrointestinal (GI) tract (FAO/WHO 2001). Health benefit reported includes (i) improved digestion of lactose and reduce intestinal bloating, flatulence and discomfort; (ii) prevention of traveler’s diarrhea; (iii) enhancing the immune system, improving resistance to infection and improving well-being; (iv) lowering serum cholesterol levels and reducing the incidence of coronary heart disease; (v) treating intractable diarrhea following antibiotic therapy; (vi) reducing allergic inflammation.
Low Calorie Sweetener
Low calorie sweetener refers to substances added to foods, drinks and nutritional supplements to provide sweet taste without calories, or at least with very few calories. Most low calorie sweeteners are several hundred times sweeter than table sugar, meaning that only small quantities need to be added to achieve a sweetening effect. These low calorie sweeteners are also referred to as high-intensity sweeteners (or intense sweeteners). Non-limiting examples of intense sweeteners include alitame, advantame, cyclamate, neotame, saccharin, sucralose, steviol glycosides and thaumatin.
Some non-sugar sweeteners are polyols, also known as “sugar alcohols”. These are, in general, less sweet than sucrose but have similar bulk properties and can be used in a wide range of food products. Sometimes the sweetness profile is “fine-tuned” by mixing with high intensity sweeteners.
Useful sugar alcohols include erythritol and xylitol, which are typically used to sweeten chewing gum, candy, fruit spreads, tooth paste, cough syrup, and other products.
Flavors
In the context of present invention a flavor refers to natural or artificial tastes, which may include fragrances and colors of the flavoring. The U. S. Code of Federal Regulations describe a natural flavorant as: the essential oil, oleoresin, essence or extractive, protein hydrolysate, distillate or any product of roasting, heating or enzymolysis, which contains the flavoring constituents derived from a spice, fruit or fruit juice, vegetable or vegetable juice, edible yeast, herb, bark, bud, root, leaf or similar plant material, meat, seafood, poultry, eggs, dairy products or fermentation products thereof, whose significant function in food is flavoring rather than nutritional. Artificial flavors include any substance, the function of which is to impart flavor, which is not derived from a spice, fruit or fruit juices, vegetable or vegetable juice, edible yeast, herb, burk, bud, root, leaf or similar plantmaterial, meat, fish, poultry, eggs, dairy products, or fermentation products thereof. Some non-limiting examples of flavors comprises of Cherry, Maple, Orange, Pineapple, Raspberry, banana, vanilla, chocolate, butterscotch, coconut, custard, cinnamon, strawberry, anish, apricot, honey, lime, coriander, lemon etc and combinations thereof.
Aroma compounds
In the context of the present invention an aroma compound, also referred to as aroma, is a chemical compound that has a smell or odor. A chemical compound has a smell or odor when it is sufficiently volatile to be transported to the olfactory system in the upper part of the nose. Generally molecules meeting this specification have molecular weights of <300. One group of aroma compounds, flavors, affects both the sense of taste and smell. Flavors are typically naturally occurring. Non-limiting examples of aroma compounds useful in context of present invention includes honeydew melon aroma, blueberry aroma, peach aroma, Strawberry aroma, raspberry aroma, cola aroma, chocolate aroma, peppermint aroma, cherry aroma, lemon aroma, lime aroma, orange aroma, vanilla aroma, tangerine aroma, liquorice aroma, apricot aroma, eucalyptus aroma, green tea aroma, ginger aroma and bilberry aroma.
Acidic regulators
In the context of the present invention acidic regulators may be included to regulate and control the pH of the composition. The acidic regulator may also be contributing to the flavor of the composition. Acidic regulators include, but are not limited to, citric acid, acetic acid, calcium acetate, lactic acid, malic acid, fumaric acid and tartaric acid.
Antioxidant Agents
The context of the present invention the term “Antioxidant” refers mainly to non-nutrient compounds, which have antioxidant capacity in vitro. Dietary antioxidant vitamins include vitamin A, Vitamin C, and Vitamin E. Preferred antioxidants include but are not limited to sodium ascorbate and alpha tocopherol.
A first aspect of the present invention concerns a nutritional composition comprising:
I. At least one live microbial organism,
II. At least one zero or low calorie sweetener,
III. At least one flavor,
IV. At least one liquid dispersion medium.
The composition may optionally comprise one or more aroma compound and/ or one or more acidity regulator and/ or one or more antioxidant agent.
Most preferably the composition is in the form of suspension, wherein the microbial organism is suspended in a dispersion medium, preferably Purified Water.
The composition of the present invention comprises at least one live microbial organism. The microbial organism (preferably a bacterium) confers a health benefit to the subject, when ingested in adequate amounts by a subject (such as in the form a formulation as described herein). It follows that the microbial organism is non-pathogenic and does not confer any harmful effect in the ingested amounts. The microbial organism is live, which may be confirmed by plating the microbial organism on a suitable medium (e.g. solidified agar in a standard sized Petri dish) and determine the number of colonies formed. The measure, Colony forming unit (or CFU), is used to quantify the amount of viable (live) microorganism in the composition (reflecting the capacity of the microorganism to replicate).
The initial colony forming units (CFU) and the continued stability and viability of the composition is influenced by variety of factors. The stability of a probiotic tested at the time of manufacturing will depend on a combination of factors. Variations in packaging, temperature and humidity will affect the viability of probiotic products before they are taken. Protective factors that help to preserve the freshness and viability of the probiotic strains in a supplement include refrigeration, resistant packaging, and storage in a cool, dry place. The CFU in the composition declines, if a probiotic composition is held in conditions that are very warm or moist. Thus, the continued stability and viability is dependent on limiting their exposure to stimulating environmental conditions such as warmth and moisture.
The colony forming units (CFU) referred to in the context of the composition of the present invention are CFU in a single dose and the CFU of the composition shortly after the preparation of the composition.
In one embodiment of the present invention, the colony forming units (CFU) of said microorganism in one dose of the composition is in the range of 10e3 (1000) to 10e12 (1000000000000), such as 10e6 (1000000) to 10e12 colony forming units. In a preferred embodiment, one dose of the composition comprises 10e7 (10000000) to 10e11 (100000000000) colony forming units, such as 10e7 to 10e10 (10000000000) colony forming units of a probiotic bacterium. In one embodiment, the at least microbial organism is present in an amount from 0.1% to 10 % w/w in said composition.
Preferably the composition aseptically filled into Blow-Fill-Seal containers, specifically tubes, more specifically six mini tubes of 5 mL each is packed in a carton, wherein each tube contains one dose of the composition, i.e., one dose of the microorganism. Unless individually packaged as preferred, the container comprising the composition will typically have a recommended serving size listed and then the colony forming units in that serving size.
Some embodiments of present invention are directed to the packaging process employing the Blow-Fill-Seal technique. Thermoplastic resin, specifically polyethylene, more specifically low density polyethylene (LDPE) is extruded as hollow tube known as Parison. The liquid LDPE might be transparent or colored for easy market differentiation. When the Parison reached to a proper length the holding jaw is closed and the parison is cut. The bottom of the parison is pinched closed while the top is held in place. The blow fill nozzle is lowered into the parison into the parison forming a seal with the neck of the mold. Sterile, filtered compressed air is blown into the parison expanding it then vented out. The sterile product, in the present context the sterile probiotic suspension, can be metered in through the fill nozzles. Separate sealing molds close to form the top and hermetically seal the container. The mold then opens and the formed, filled and sealed containers are conveyed out of the machine.
In one embodiment of present invention, the at least one microbial organism is a gram positive bacterium. In a further embodiment, the at least one microbial organism is selected from the list consisting of Bacillus clausii, Bacillus sporogensis, Bifidobacterium adolescentis, Fidobacteriumanimalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum, Lactobacillus acidophilus, Bifidobacterium lactis, Bifidobacterium infantis, Lactobacillus amylovorus, Lactobacillus casei, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus salivarius, Streptococcus thermophilus, Streptococcus salivarius, Streptococcus oralis, Streptococcus uberis, Streptococcus rattus, Escherichia coli, Bacillus coagulans, Bacillus lansii, Saccharomyces cerevisiae, Saccharomyces boulardii and combinations thereof. In even more preferred embodiment, the at least one microbial organism is Bacillus clausii.
In order to obtain the desired health benefit to the subject, it may be advantageous to include more than one microorganism in the composition. Thus the composition may comprise more than one species/ strain of microorganism, such as two, three, four, five or a higher plurality of species/ strains of microorganism. In one embodiment, the composition comprises Bacillus clausii and Bacillus sporogensis.
The composition of present invention further comprises at least one low calorie sweetener. In a further embodiment, the low calorie sweetener is selected from the list consisting of alitame, ace sulfate potassium (Ace K), aspartame, advantame, cyclamate, neotame, saccharin, sucralose, steviol glycosides and thaumatin. In a preferred embodiment, the composition comprises a low calorie sweetener in the form of sucralose. The inventors have found the rounded sweetness of sucralose combined with a flavor, such as orange flavor, chocolate flavor or combination thereof, and the formulation of the composition as suspension that gives a pleasant experience upon ingestion contributing to an improved mouth feel specifically for children.
The amount of low calorie sweetener may vary depending on the sweetener(s) used. In one embodiment, the low calorie sweetener is present in an amount from 0.01% to 90% w/w in said composition.
Non limiting examples of the composition of the present invention include:
A nutritional composition comprising:
1. 0.1% to 10% w/w of at least one live microbial organism,
2. 0.01% to 90% w/w of at least one low or zero calorie sweetener,
3. 0.01% to 20% flavors,
4. Rest is liquid dispersion medium.
The nutrition composition optionally may comprise other pharmaceutically acceptable ingredients such as, antioxidant agents, aroma compounds, flavors, acidity regulators and combinations thereof.
In a preferred embodiment, the nutritional composition comprises or consists essentially of:
1. 1% to 5% w/w of a mixture of Bacillus clausii and Bacillus sporogensis.
2. 0.01% to 10% w/w sucralose.
3. 1% to 3% flavors,
4. Rest is Purified Water.
The composition of present invention which is essentially a suspension is aseptically filled in to a Blow-Fill-Seal tube in a sterile condition to maintain the integrity, stability and viability of the composition.
When describing the embodiments of the present invention, the combinations and permutations of all possible embodiments have not been explicitly described. Nevertheless the mere fact that certain measures are recited in mutually different dependent claims or described in different embodiments does not indicate that a combination of these measures cannot be used to advantage. The present invention envisages all possible combinations and permutations of the described embodiments.
The term “comprising”, “comprise” and “comprises” herein are intended by the inventors to be optionally substitutable with the terms “consisting of”, “consist of” and “consists of”, respectively for every embodiment disclosed herein.
Preparation of composition of present invention
One aspect of the present invention provides a method for preparing a composition containing a microbial organism, said method comprising the steps of:
(a) Aseptic mixing at least one microbial organism with at least one low or zero calorie sweeteners, at least one flavor, at least one liquid dispersion medium and optionally one or more aroma compound, optionally one or more antioxidant agent.
(b) Filling the suspension in a compatible container maintaining the aseptic condition and sealing.
Description:
Blending:
Based on CFU count the container is taken from 2-8º C and the spores pellet issuspended in required quantity of purified water under LAFU aseptically to make 0.4 billionspores in one ml and were transferred to Blender. The blender is allowed to mix at 100±10rpm for 30 min to form uniform solution. The blended mixture vessel transferred to thefilling area.
Filling and sealing:
The total Batch quantity to be filled is filled in a formulation vesseladjacent to BFS filling machine and required quantity (equal to 5.2ml ± 0.1 each) ofBacillus clausii spores suspension is dispensed with the help of the needlesprovided to the machine in to each tube automatically and sealed simultaneously. Thermoplastic resin, specifically polyethylene, more specifically low density polyethylene (LDPE) is extruded as hollow tube known as Parison. The liquid LDPE might be transparent or colored for easy market differentiation. When the Parison reached to a proper length the holding jaw is closed and the parison is cut. The bottom of the parison is pinched closed while the top is held in place. The blow fill nozzle is lowered into the parison forming a seal with the neck of the mold. Sterile, filtered compressed air is blown into the parison expanding it then vented out. The sterile product, in the present context the sterile probiotic suspension, can be metered in through the fill nozzles. Separate sealing molds close to form the top and hermetically seal the container. The mold then opens and the formed, filled and sealed containers are conveyed out of the machine.
Uses of nutritional composition of the present invention
The composition of the present invention might be used to improve the health of a subject. The health benefit might be in the form of (i) improved digestion of lactose and reduce intestinal bloating, flatulence and discomfort, (ii) prevention of traveler’s diarrhea; (iii) enhancing the immune system, improving resistance to infection and improving well-being; (iv) lowering serum cholesterol levels and reducing the incidence of coronary heart disease, or (v) treating intractable diarrhea following antibiotic therapy; (vi) reducing allergic inflammation. These health benefits might be associated with the colonization of the probiotic microorganism in the gut of the subject.
One aspect of present invention therefore provides a method for stimulating of the intestinal barrier, providing antimicrobial substances of the gut, stimulating gut transit, and/ or stimulating the immune system of a subject, preferably for children, said method comprising the administering an effective amount of the present invention to the subject, specifically children, in the need thereof.
Example 1:
The method for preparing the composition of the present invention containing a microbial organism generally comprises the steps, said method comprising the steps of:
• Weighing off of raw materials according to a master recipe.
• The raw materials are mixed together in specific order to ensure homogeneity of the finished blend. Mixing time and sequence is specified to minimize the loss of viable probiotic cells during the process.
• Thermoplastic resin, specifically polyethylene, more specifically low density polyethylene (LDPE) is extruded as hollow tube known as Parison. The liquid LDPE might be transparent or colored for easy market differentiation. When the Parison reached to a proper length the holding jaw is closed and the parison is cut. The bottom of the parison is pinched closed while the top is held in place. The blow fill nozzle is lowered into the parison forming a seal with the neck of the mold. Sterile, filtered compressed air is blown into the parison expanding it then vented out. The sterile product, in the present context the sterile probiotic suspension, can be metered in through the fill nozzles. Separate sealing molds close to form the top and hermetically seal the container. The mold then opens and the formed, filled and sealed containers are conveyed out of the machine.
Stability Study
Accelerated Stability study has been performed at 40°C ± 2°C temperature and 75% ± 5% relative humidity and the suspension is stable for six months in this accelerated condition.
S. No Test Specifications Results
Initial 1st Month 2nd Month 3rd Month 6th Month

1. Description Creamy White Suspension Creamy White Suspension Creamy White Suspension Creamy White Suspension Creamy White Suspension Creamy White Suspension
2. pH 6.5-7.5 6.88 6.86 6.85 6.83 6.83
3. Extractable Volume 5.0±0.1 mL 5.1 ml 5.1 ml 5.1 ml 5.1 ml 5.1 ml
4. Taste Sweet orange flavor Complies Complies Complies Complies Complies
5. Identification Tests
5.1 Physical Tests
Gram staining Gram Positive rod shaped bacteria Complies Complies Complies Complies Complies
Spores staining Ellipsoidal green stained spores Complies Complies Complies Complies Complies
5.2 Biochemical tests
Test for alkalinity Growth should be observed between pH 8.0 -10.0 Growth was observed between pH 8.0-10.0 Growth was observed between pH 8.0-10.0 Growth was observed between pH 8.0-10.0 Growth was observed between pH 8.0-10.0 Growth was observed between pH 8.0-10.0
Test for hydrolysis of starch Positive Positive Positive Positive Positive Positive
Test for hydrolysis of casein Positive Positive Positive Positive Positive Positive
Test for hydrolysis of Gelatin Positive Positive Positive Positive Positive Positive
Phenylalanine deaminase test Negative Negative Negative Negative Negative Negative
Tween 20/80 hydrolysis test Negative Negative Negative Negative Negative Negative
NaCl tolerance test Should be 2-8% Growth was observed between 2-8% of Nacl concentration Growth was observed between 2-8% of Nacl concentration Growth was observed between 2-8% of Nacl concentration Growth was observed between 2-8% of Nacl concentration Growth was observed between 2-8% of Nacl concentration
6. In vitro test
Gastric acid resistance test Positive Positive Positive Positive Positive Positive
6.2 Antibiotic resistance test
Streptomycin test Positive Positive Positive Positive Positive Positive
Tetracycline test Positive Positive Positive Positive Positive Positive
Rifampin test Positive Positive Positive Positive Positive Positive
Chloramphenicol test Positive Positive Positive Positive Positive Positive
7. CFU NLT 400 Million per 1 ml 425 Million per 1 ml 420 Million per 1 ml 420 Million per 1 ml 415 Million per 1 ml 410 Million per 1 ml
8. Impurities
Arsenic NMT 1ppm ‹1 ppm ‹1 ppm ‹1 ppm ‹1 ppm ‹1 ppm
Heavy Metals NMT 20ppm ‹20 ppm ‹20 ppm ‹20 ppm ‹20 ppm ‹20 ppm
9. Microbial Limit Test
Escherichia coli Absent Absent Absent Absent Absent Absent
Staphylococcus aureus Absent Absent Absent Absent Absent Absent
Salmonella Species. Absent Absent Absent Absent Absent Absent
Pseudomonas aeruginosa Absent Absent Absent Absent Absent Absent
Result: Pass/Fail Pass Pass Pass Pass Pass

The systems and methods of the present invention may be embodied in other specific forms without departing from the teachings or essential characteristics of the invention.
The foregoing embodiments and advantages are merely exemplary and are not to be construed as limiting the scope of the present invention. The description of the exemplary embodiments of the present invention is intended to be illustrative and not to limit the scope of the invention. Various modifications, alterations and variations, which are apparent to a person skilled in the art, are intended to fall within the scope of the invention.