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WO2024115675A1 - Encapsulated core material - Google Patents

Encapsulated core material Download PDF

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Publication number
WO2024115675A1
WO2024115675A1 PCT/EP2023/083775 EP2023083775W WO2024115675A1 WO 2024115675 A1 WO2024115675 A1 WO 2024115675A1 EP 2023083775 W EP2023083775 W EP 2023083775W WO 2024115675 A1 WO2024115675 A1 WO 2024115675A1
Authority
WO
WIPO (PCT)
Prior art keywords
oil
particulate material
core material
layer
lipid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP2023/083775
Other languages
French (fr)
Inventor
Asger SMIDT-JENSEN
Katrine Kiib RØRBY
Hanne Bjørn NEDERGAARD
Lisa TAMMI
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Mello Aps
Original Assignee
Mello Aps
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Mello Aps filed Critical Mello Aps
Publication of WO2024115675A1 publication Critical patent/WO2024115675A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02—Making microcapsules or microballoons
    • B01J13/20—After-treatment of capsule walls, e.g. hardening
    • B01J13/22—Coating
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • A23P10/35—Encapsulation of particles, e.g. foodstuff additives with oils, lipids, monoglycerides or diglycerides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/0241—Containing particulates characterized by their shape and/or structure
    • A61K8/0245—Specific shapes or structures not provided for by any of the groups of A61K8/0241
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34—Alcohols
    • A61K8/342—Alcohols having more than seven atoms in an unbroken chain
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A61K8/361—Carboxylic acids having more than seven carbon atoms in an unbroken chain; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/99—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from microorganisms other than algae or fungi, e.g. protozoa or bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
    • B01J13/02—Making microcapsules or microballoons
    • B01J13/06—Making microcapsules or microballoons by phase separation
    • B01J13/14—Polymerisation; cross-linking
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/10—General cosmetic use
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/41—Particular ingredients further characterized by their size
    • A61K2800/412—Microsized, i.e. having sizes between 0.1 and 100 microns
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/60—Particulates further characterized by their structure or composition
    • A61K2800/65—Characterized by the composition of the particulate/core
    • A61K2800/652—The particulate/core comprising organic material
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/5005—Wall or coating material
    • A61K9/5015—Organic compounds, e.g. fats, sugars

Definitions

  • the present invention relates to a method for providing a microencapsulated core material.
  • the present invention relates to a method of providing a microencapsulated core material, and the microencapsulated core material as such, which may safely and efficiently deliver probiotics and other sensitive particulate and non-particulate materials to target locations in the body.
  • the human digestive tract comprises trillions of bacteria, many of which help digest food and fight off harmful bacteria. Studies have shown that some of these bacteria may influence a range of diseases, for better or worse.
  • microbiome manipulating these bacterial populations in the gastrointestinal tract, known collectively as the microbiome, could improve human health.
  • Varies strategies of manipulating the microbiome have been suggested and may be divided into two main groups, the prebiotic strategies and the probiotic strategies.
  • the prebiotic strategies relate to compounds in food that may induce the growth or activity of beneficial microorganisms, such as bacteria and fungi, in the gastrointestinal tract, where the prebiotics may be altering the composition of organisms in the microbiome.
  • Dietary prebiotics are typically nondigestible fibre compounds that pass undigested through the upper part of the gastrointestinal tract and stimulate the growth or activity of advantageous bacteria in the colon by acting as substrates for them.
  • the probiotic strategies relate to live microorganisms capable of or intended to provide health benefits when consumed.
  • the health benefits may be provided by improving or restoring the gut microbiota of the mammal consuming the probiotic.
  • Probiotics are considered generally safe to consume.
  • One of the challenges of manipulating the bacterial population of beneficial bacteria in the intestine, in particular using probiotic, thus live microorganisms, may be to deliver large numbers of beneficial bacterial to the gut, where the microorganisms delivered must be live microorganisms, and therefore they need to survive the passage of the bile salts and acidic conditions of the stomach.
  • an object of the present invention relates to a method for providing a microencapsulated core material.
  • the present invention relates to a method of providing a microencapsulated core material, and the microencapsulated core material as such, which may safely and efficiently deliver probiotics and other sensitive particulate and non-particulate materials to target locations in the body, in particular the intestine.
  • one aspect of the invention relates to a particulate material comprising an encapsulated core material, wherein the encapsulated core material comprises an oil layer, wherein the oil layer is surrounded by a lipid layer.
  • Another aspect of the present invention relates to a method for providing a particulate material, the method comprises the steps of:
  • step (iv) Combining the encapsulated core material comprising an oil layer (from step (ii) with the molten lipid composition (from step (iii)), for providing a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
  • Yet another aspect of the present invention relates to a food product or a feed product comprising the particulate material according to the present invention.
  • Still another aspect of the present invention relates to a beverage comprising a particulate material according to the present invention.
  • a further aspect of the present invention relates to a medical product for use as a medicament.
  • An even further aspect of the present invention relates to a medical product for use in the treatment of gut diseases, immune diseases, skin diseases, and/or oral diseases.
  • Figure 1 shows a coating device (1) according to the present invention.
  • the coating device (1) a chamber (2), one material inlet (3), 3 coating inlet (4), and a particulate material outlet (5).
  • the material inlet (3) comprises at a material nozzle and each 3 coating inlets (4) comprises at least one coating nozzle (7) and each of the 3 coating nozzles (7) are angled relative to the material nozzle (6). This angle provides a spray zone (14) where the 3 clouds of coating agents are contacting the cloud of the material to be coated (and the particles in the cloud of the material to be coated) from different sides resulting in an improved coating of the particulate material and having a more complete coating of the particulate material.
  • the material inlet (3) may comprise a material feed (8) and a feed of compressed gas (9) and the 3 coating inlets (4) comprises a coating feed (10) and a feed of compressed gas (9).
  • the material inlet (3) is in fluid connection with a tank (11) comprising the material to be coated and the 3 coating inlets (4) are in fluid connection with a coating agent tank (12) comprising the coating agent.
  • the material inlet (3) and/or in particular the 3 coating inlets (4) are providing with means for heating.
  • the material nozzle (6) may provide a cloud of material to be coated with a density of particles which may be customized and the 3 coating nozzles (7) may provide clouds of coating agent with a density of particles which is customized, in such a way that coating of the individual particles may be provided.
  • the particulate material obtained from the particulate material outlet (5) may be send to a separation process using a device like a cyclone (13).
  • the device may optionally be equipped with means for cooling.
  • Figure 2 shows various stages of solidification and the stage preferably reached by the present invention.
  • the first stage (fig. 2a) shows the contact between a material to be coated and a coating material and how the coating agent is accumulated on the surface of the material to be coated, since the coating agent may be solidified before reaching the material to be coated.
  • figure 2b it is shown the proper contact between a material to be coated and a coating material and how the coating agent stick to the surface of the material to be coated and spreads before being solidified. This is possible because the coating agent reaches the material to be coated before the coating agent is solidified.
  • Figure 3 shows a microscopy image (polarized light microscopy of coated particles according to the present invention, with a shell (lipid layer) visible as lighter part surrounding the darker core material provided with an oil layer.
  • a shell lipid layer
  • the aim of the present invention relates to microencapsulation of sensitive materials, such as probiotics.
  • sensitive materials such as probiotics.
  • the inventor of the present invention found a method for providing a particulate material comprising an encapsulated core material.
  • the particulate material may be a powdered particulate material.
  • the core material may comprise some water, in particular when the core material may be a microorganism, such as a lactic acid bacteria or a probiotic microorganism, the particulate material may have a moisture content below 5% (w/w), preferably below 3% (w/w), even more preferably below 1% (w/w), even more preferably no (or substantially no) moisture.
  • the core material may be a microorganism, such as a lactic acid bacteria or a probiotic microorganism
  • the particulate material may have a moisture content below 5% (w/w), preferably below 3% (w/w), even more preferably below 1% (w/w), even more preferably no (or substantially no) moisture.
  • Microencapsulation techniques may be a process in the live microorganism may be surrounded by a coating to give small capsules.
  • the present invention relates to a novel microencapsulation to deliver live probiotics safely and efficiently to the ileal intestine and/or to the large intestine, preferably, to the large intestine.
  • the present invention relates to a particulate material comprising an encapsulated core material including an oil-based true-core, preferably comprising one or more microorganism, and surrounded by a lipid-based true shell that may be highly tolerant to low gastric pH conditions and/or moisture.
  • an oil-based true-core preferably comprising one or more microorganism
  • a lipid-based true shell that may be highly tolerant to low gastric pH conditions and/or moisture.
  • the particulate material comprising an encapsulated core material may have a high- release rate in response to stimuli from the body when delivered to the microbiome in the gastrointestinal track.
  • a preferred embodiment of the present invention relates to a particulate material comprising an encapsulated core material, wherein the encapsulated core material may comprise an oil layer, wherein the oil layer may be surrounded by a lipid layer.
  • the particulate material may be a small, often very small particles which may comprise a cores material.
  • the particulate material may have a mean particle size (D50) less than 1000pm, such as less than 750pm, e.g. less than 500pm, such as less than 300pm, e.g. less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
  • D50 mean particle size
  • the particulate material may have a mean particle size (D90) less than 1000pm, such as less than 750pm, e.g. less than 500pm, such as less than 300pm, e.g. less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
  • D90 mean particle size
  • the particulate material may vary greatly in size, shape, dispersible, and digestibility.
  • the core material according to the present invention may comprise a sensitive material.
  • the sensitive material may be sensitive to the influence of chemicals, enzymes, oxygen, light, or a combination hereof.
  • the core material may be a microorganism, food ingredients, cosmetic ingredients, pharmaceutical component, enzymes, dyes, vitamins, cells or other materials.
  • the core material may not be an oil, such as a fish oil or a plant oil.
  • the core material may comprise a microorganism.
  • the sensitive material may be a microorganism.
  • the microorganism may be a bacterium, a fungus, a yeast, or a virus.
  • the microorganism may be a viable microorganism.
  • the viable microorganisms may be live microorganisms and/or viable spores.
  • the core material may comprise one or more probiotic microorganism.
  • the probiotic microorganism may relate to live microorganisms which when administered in adequate amounts to a human or animal may confer a health benefit.
  • the microorganism may be a microorganism which is capable of introducing, supporting or boosting the development of a beneficial gut microflora.
  • the core material may comprise one or more bacterial strain.
  • the one or more bacterial strain may include one or more lactic acid bacterial strains.
  • the one or more bacterial strain may consist essentially of one or more lactic acid bacterial strains.
  • the one or more lactic acid bacterial strains may comprise Lactobacillus, Bifidobacterium or a combination hereof.
  • the core material may comprise a concentration of live or viable microorganism, in particular live or viable bacterial, in the range of 10 3 -10 15 CFU/g, such as in the range of 10 5 -10 14 CFU/g, e.g. in the range of 10 6 -10 12 CFU/g, such as in the range of IO 7 - 10 11 CFU/g, e.g. in the range of 10 8 -10 10 CFU/g.
  • CFU relates to colony-forming unit and may be a unit which estimates the number of microbial cells (in particular bacteria) in a sample that are viable and thus able to multiply under controlled conditions.
  • the microorganisms provided for the core material may be provided in moisten state, in dry state or in frozen state before suspended in the component.
  • the lipid layer and the oil layer according to the present invention may both comprises fatty acids.
  • One difference between the lipid layer and the oil layer according to the present invention may be that at room temperature the lipid layer may be solid whereas oil layer may be liquid.
  • room temperature according to the present invention may be a temperature in the range of 18-25°C.
  • the oil layer may comprise an oil component.
  • the oil component may comprise a single oil component of a combination of two or more oils compounds or a combination of oil and lipid compounds.
  • the oil component may comprise a vegetable oil.
  • the oil component may be liquid at a temperature below 30°C, e.g. the oil component may be liquid at a temperature below 25°C, e.g. the oil component may be liquid at a temperature below 20°C, e.g. the oil component may be liquid at a temperature below 15°C, e.g. oil component may be liquid at a temperature below 10°C, such as at a temperature in the range of 5-35°C, e.g. in the range of 10-30°C, such as in the range of 15-25°C, e.g. of about 25°C.
  • the oil component and/or the oil layer may comprise palm oil, coconut oil, canola oil, soybean oil, corn oil, rapeseed oil, cottonseed oil, olive oil, sunflower oil or a combination hereof.
  • the oil component and/or the oil layer may comprise coconut oil, rapeseed oil, sunflower oil, or a combination hereof.
  • the core material may be encapsulated by the oil layer by mixing the core material according to the present invention with the oil component creating a core material comprising an oil layer.
  • the oil layer may provide a coherent layer, preferably a coherent thin layer, of oil component surrounding the core material.
  • the oil layer may surround the core material as a hole uniform element, and/or the oil layer may surround individual parts of the core material making up the hole uniform element.
  • the lipid layer may provide at least 20% encapsulation efficiency, such as at least 30% encapsulation efficiency, e.g. at least 40% encapsulation efficiency, such as at least 50% encapsulation efficiency, e.g. at least 60% encapsulation efficiency, such as at least 80% encapsulation efficiency, e.g. at least 90% encapsulation efficiency, such as at least 95% encapsulation efficiency, e.g. at least 98% encapsulation efficiency.
  • encapsulation efficiency such as at least 30% encapsulation efficiency, e.g. at least 40% encapsulation efficiency, such as at least 50% encapsulation efficiency, e.g. at least 60% encapsulation efficiency, such as at least 80% encapsulation efficiency, e.g. at least 90% encapsulation efficiency, such as at least 95% encapsulation efficiency, e.g. at least 98% encapsulation efficiency.
  • the term “encapsulation efficiency" may relate to the amount of core material maintained in the particulate material when exposed to water.
  • the exposure to water may be washing with water.
  • the washing with water may include a vigorously mixing and/or stirring.
  • the mean particle size (D50) of the core material surrounded by the oil layer may be less than 100 pm, such as less than 75 pm, e.g. less than 60 pm, such as less than 50 pm, e.g. less than 40 pm, such as less than 30 pm, e.g. less than 20 pm, such as less than 10 pm, e.g. in the range of 5-125 pm, such as in the range of 10-100 pm, e.g. in the range of 20-75 pm, such as in the range of 30-60 pm, e.g. in the range of 40-50 pm.
  • the mean particle size (D90) of the core material surrounded by the oil layer may be less than 100 pm, such as less than 75 pm, e.g. less than 60 pm, such as less than 50 pm, e.g. less than 40 pm, such as less than 30 pm, e.g. less than 20 pm, such as less than 10 pm, e.g. in the range of 5-125 pm, such as in the range of 10-100 pm, e.g. in the range of 20-75 pm, such as in the range of 30-60 pm, e.g. in the range of 40-50 pm.
  • the core material may be suspended in the oil component (or combination of oil components) to a substantially homogenous mixture.
  • the core material may be suspended in the oil component (or combination of oil components) to provide a substantially homogenous mixture at a temperature below 30°C, e.g. the oil component may be liquid at a temperature below 25°C, e.g. the oil component may be liquid at a temperature below 20°C, e.g. the oil component may be liquid at a temperature below 15°C, e.g. oil component may be liquid at a temperature below 10°C, such as at a temperature in the range of 5-35°C, e.g. in the range of 10-30°C, such as in the range of 15-25°C, e.g. of about 25°C.
  • the core material according to the present invention may constitute in the range of 1-50% (w/w) of the encapsulated core material comprises an oil layer, such as in the range of 10-45% (w/w), e.g. in the range of 15-40% (w/w), such as in the range of 20-35% (w/w), e.g. in the range of 25-30% (w/w).
  • the encapsulated core material comprising the oil layer may be surrounded by a lipid layer.
  • the lipid layer may be provided by a lipid compound.
  • the lipid compound may comprise a single lipid compound, a combination of different lipid compounds, a combination of a single lipid compound and an oil, or a combination of two or more different lipid compounds and an oil.
  • the lipid compound may a solid at a temperature below 25°C, e.g. the oil component may be a solid at a temperature below 20°C, e.g. the oil component may be a solid at a temperature below 15°C, e.g. oil component may be a solid at a temperature below 10°C, such as at a temperature in the range of 5-35°C, e.g. in the range of 10-30°C, such as in the range of 15-25°C, e.g. of about 25°C.
  • the lipid compound may be liquid at a temperature below 70°C, e.g. the oil component may be liquid at a temperature below 65°C, e.g. the oil component may be liquid at a temperature below 60°C, e.g. the oil component may be liquid at a temperature below 55°C, e.g. the oil component may be liquid at a temperature below 50°C, e.g. the oil component may be liquid at a temperature below 40°C, e.g. the oil component may be liquid at a temperature below 30°C, e.g. the oil component may be liquid at a temperature below 20°C, such as at a temperature in the range of 20-70°C, e.g. in the range of 40-65°C, such as in the range of 50-60°C, e.g. in the range of 52-58°C.
  • solid relates to a compound that has a three-dimensional stable structure or shape and does not deform by the action of gravity.
  • the lipid layer may comprise a fatty acid, a fatty alcohol, a wax, a sterol, a phosphor lipid, or a combination hereof.
  • the lipid layer may comprise a fatty acid, a fatty alcohol, or a combination hereof.
  • the fatty acid or fatty alcohol may preferably be a saturated fatty alcohol or a saturated fatty acid and/or the fatty acid or fatty alcohol may comprise a carbon chain length ranging from C12-C22, such as C12-C20, e.g. C12-C18.
  • the lipid layer may comprise the combination of a fatty acid and a fatty alcohol.
  • the presence of fatty alcohol in the lipid layer may be preferred over the presence and/or concentration of fatty acid.
  • the fatty acid may be selected from lauric acid, myristic acid, or a combination hereof.
  • the fatty alcohol may be selected from cetyl alcohol, stearyl alcohol, tetradecanol, or a combination hereof.
  • the lipid layer may comprise stearyl alcohol in combination with lauric acid, myristic acid, palmitic acid or a combination hereof.
  • the content of stearyl alcohol may represent at least 20% (w/w) of the lipid layer, such as at least 30% (w/w) of the lipid layer, e.g. at least 40% (w/w) of the lipid layer, such as at least 50% (w/w) of the lipid layer, e.g. at least 60% (w/w) of the lipid layer, such as at least 70% (w/w) of the lipid layer, e.g. at least 80% (w/w) of the lipid layer, such as at least 90% (w/w) of the lipid layer, e.g. at least 95% (w/w) of the lipid layer, such as about 98% (w/w) of the lipid layer, e.g. about
  • the content of lauric acid may represent in the range of 0.1-10% (w/w), such as in the range of 0.2-5% (w/w), e.g. in the range of 0.3- 2% (w/w), such as in the range of 0.5-1% (w/w).
  • the content of myristic acid may represent in the range of 0.1-10% (w/w), such as in the range of 0.2-5% (w/w), e.g. in the range of 0.3-2% (w/w), such as in the range of 0.5-1% (w/w).
  • the temperature of the lipid composition when applied to the encapsulated core material comprising an oil layer may be above the melting point of the lipid composition.
  • the melting point of the lipid composition may be the temperature and/or energy level necessary for changing the state of the lipid composition from a solid state to a liquid state
  • the melting point of the lipid composition may be dependent on the composition of the lipid composition.
  • the temperature of the lipid composition, before contacting the lipid composition with the encapsulated core material comprising an oil layer may be set in such a way that the lipid composition may be liquid at the time of contact with the encapsulated core material comprising an oil layer may be.
  • the lipid composition may be surrounding the encapsulated core material comprising an oil layer and the lipid layer may be allowed to solidify, creating a lipid layer.
  • a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer, may be provided.
  • solidify means that the lipid composition (or the lipid layer) changes state from a liquid state to a solid state.
  • the temperature of the lipid composition at the time of contacting the encapsulated core material comprising an oil layer may be above the melting point of the lipid composition.
  • the lipid composition has a temperature above 25°C, such as above 30°C, e.g. above 35°C, such as above 40°C, e.g. above 45°C, such as above 50°C, e.g. above 55°C, such as above 60°C, e.g. above 65°C, such as in the range of 25-70°C, e.g. in the range of 30-65°C, such as in the range of 35- 60°C, e.g. in the range of 40-55°C, such as in the range of 45-50°C.
  • the temperature of the lipid composition may be set at a temperature just slightly above the melting point of the lipid composition.
  • the temperature of the lipid composition may be set at a temperature in the range of l-20°C above the melting point of the lipid composition, such as in the range of 1-15°C above the melting point of the lipid composition, e.g. in the range of 2-10°C above the melting point of the lipid composition, such as in the range of 3-8°C above the melting point of the lipid composition, e.g. in the range of 4-6°C above the melting point of the lipid composition.
  • the encapsulated core material comprising an oil layer and surrounded by a lipid layer may be subjected to cooling in order to solidify the lipid layer around the encapsulated core material comprising an oil layer providing the particulate material according to the present invention.
  • the particulate material according to the present invention may maintain at least 10% of the core material, such as at least 20% of the core material, e.g. at least 30% of the core material, such as at least 40% of the core material, e.g. at least 50% of the core material, such as at least 60% of the core material, e.g. at least 70% of the core material, such as at least 80% of the core material, e.g.
  • At least 90% of the core material, encapsulated in the oil layer and the lipid layer after a period of at least 10 minutes in a test solution such as for a period of at least 30 minutes, e.g. for at least 45 minutes, such as for a period of at least 60 minutes, e.g. for at least 2 hours, such as for a period of at least 6 hours, e.g. for at least 12 hours, such as for a period of at least 24 hours, e.g. for at least 2 days, such as for a period of at least 5 days, e.g. for at least 10 days, such as for a period of at least 20 days, e.g. for at least 30 days, such as for a period of at least 40 days, e.g. for at least 50 days, such as for a period of at least 60 days, e.g. for at least 70 days.
  • a test solution such as for a period of at least 30 minutes, e.g. for at least 45 minutes, such as for a period
  • the particulate material may comprise a core material comprising (consisting essentially of) viable or live microorganisms, in particular one or more probiotics, and at least 10% of the microorganisms, in particular the probiotics, such as at least 20%, e.g. at least 30%, such as at least 50%, e.g. at least 60%, such as at least 70%, e.g. at least 80% of the core material, such as at least 90% of the core material, e.g. at least 95%, of the microorganisms, in particular one or more probiotics, remains alive or viable, after storage for a period of at least 10 minutes in a test solution, such as for a period of at least 30 minutes, e.g.
  • the core material comprises one or more viable microorganisms and wherein one or more microorganisms show a reduction in viability of at most 3 LOG over a period of storage of 12 months at room temperature.
  • the particulate material comprising a core material comprising (consisting essentially of) one or more viable microorganisms according to the present invention and wherein the one or more viable microorganisms show a reduction in viability of at most 3 LOG, such as a reduction of at the most 2 LOG, e.g. a reduction of at the most 1 LOG reduction, such as a reduction of at the most 0.5 LOG, e.g. a reduction of at the most 0.1 LOG reduction over a period of storage of 12 months at room temperature.
  • the test solution may be selected from water, aqueous acidic environment, or an aqueous environment comprising bile salts and or bile acids.
  • the surface of the encapsulated core material comprising an oil layer of the particulate material may be covered or partly covered by the lipid layer.
  • the encapsulated core material comprising an oil layer of the particulate material may be at least 40% covered by the lipid layer, such as at least 50% covered, e.g. at least 60% covered, such as at least 70% covered, e.g. at least 80% covered, such as at least 85% covered, e.g. at least 90% covered, such as at least 95% covered.
  • the effect of the lipid layer may be determined by the efficiency to retain a core material in the particulate material.
  • the larger part of the particulate material being covered by the lipid layer i.e. the higher the coverage), the higher the amount of the core material may be retained in the particle material and the less susceptible the particulate material is to obtain material from the outside of the particulate material.
  • the particulate material according to the present invention may provide a retention of a trace element of 70% or above, such as 75% or above, e.g. 80% or above, such as 85% or above, e.g. 90% or above, such as 92% or above, e.g. 94% or above, such as 96% or above, e.g. 98% or above.
  • the particulate material according to the present invention may provide a retention of the core material of 70% or above, such as 75% or above, e.g. 80% or above, such as 85% or above, e.g. 90% or above, such as 92% or above, e.g. 94% or above, such as 96% or above, e.g. 98% or above.
  • the encapsulation efficiency and/or the lipid coverage or the retention properties of the particulate material may be determined by a diffusion/retention test using a trace element.
  • the trace element may be Patent Blue Colorant, Oil Red O or the like.
  • the outer surface of a particulate material comprising a microbial core material may comprise less than 10 6 CFU/g particulate material, such as less than 10 5 CFU/g particulate material, e.g. less than 10 4 CFU/g particulate material, such as less than 10 3 CFU/g particulate material, e.g. less than 10 2 CFU/g particulate material.
  • a preferred embodiment of the present invention relates to a method for providing a particulate material, the method comprises the steps of:
  • step (iv) Combining the encapsulated core material comprising an oil layer (from step (ii) with the molten lipid composition (from step (iii)), providing a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
  • agitation may be provided during mixing in step (ii).
  • step (iv)) may be performed in a coating device, the coating device comprising a chamber, wherein the chamber comprising at least one material inlet and at least one coating inlet and a particulate material outlet.
  • the at least one material inlet and/or the at least one coating inlet may be placed in the upper part of the chamber.
  • each of the at least one material inlet may comprise at least one material nozzle and/or wherein each the at least one coating inlet comprises at least one coating nozzle.
  • the at least one coating nozzle may be angled relative to the material nozzle.
  • the material nozzle of the material inlet may be pointing vertically downwards, or substantially vertically downwards.
  • the coating nozzle may be angled by 1-179° relative to the material nozzle, such as by 2- 150°, e.g. by 3-125°, such as by 4-100°, e.g. by 5-75°, such as by 6-50°, e.g. by 7-35°, such as by 8-25°, e.g. by 9-20°, such as by 10-18°, e.g. about 15° .
  • the at least one material inlet and/or the at least one coating inlet are providing with means for heating.
  • the at least one material inlet may be in fluid connection with a tank comprising the material to be coated.
  • the at least one coating inlet may be in fluid connection with a coating agent tank comprising the coating agent.
  • the chamber may comprise at least two coating inlets, such as at least 3 coating inlets, e.g. at least 4 coating inlets, such as at least 5 coating inlets, e.g. at least 6 coating inlets, such as at least 7 coating inlets, e.g. at least 8 coating inlets.
  • the chamber comprising of the coating device is provided with one material inlet and at least one coating inlet, e.g. at least two coating inlets, such as at least 3 coating inlets, e.g. at least 4 coating inlets, such as at least 5 coating inlets, e.g. at least 6 coating inlets, such as at least 7 coating inlets, e.g. at least 8 coating inlets.
  • at least two coating inlets such as at least 3 coating inlets, e.g. at least 4 coating inlets, such as at least 5 coating inlets, e.g. at least 6 coating inlets, such as at least 7 coating inlets, e.g. at least 8 coating inlets.
  • the two or more coating inlets are surrounding the material inlet to improve the distribution of the lipid layer on the surface of the encapsulated core material comprising and an oil layer, and at the same time keeping the mean particle size low.
  • the method of coating the encapsulated core material comprising and an oil layer with the lipid layer may comprise the steps of:
  • the combining may involve putting or applying the molten lipid composition on the top or the surface of the encapsulated core material comprising an oil layer, resulting in the particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
  • the particles of the encapsulated core material comprising an oil layer and a lipid layer may comprise a mean particle size (D50) of less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
  • D50 mean particle size
  • the particles of the encapsulated core material comprising an oil layer and a lipid layer may comprise a mean particle size (D90) of less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
  • D90 mean particle size
  • the mean particle size or the average particles size may be determined by sieving, sedimentation electrozone testing, automated microscopy and/or by laser diffraction.
  • the mean particle size may be selected according to the use or application of the particulate material, or the encapsulated core material comprising an oil layer and surrounded by a lipid layer.
  • the lipid layer may be allowed to solidify, e.g. by subjecting the encapsulated core material comprising an oil layer and surrounded to e.g. cooling, providing a particulate material according to the present invention.
  • the particulate material according to the present invention may be a substantially spherical particulate material.
  • an embodiment of the present invention the encapsulated core material comprising an oil layer and surrounded by a lipid layer (obtained in step (iv)) may be combined with the molten lipid composition.
  • the encapsulated core material comprising an oil layer and surrounded by a lipid layer may be combined with the molten lipid composition to increase the thickness of the lipid layer of the encapsulated core material comprising an oil layer and surrounded by a lipid layer.
  • the step of combining the encapsulated core material comprising an oil layer and surrounded by a lipid layer with the molten lipid composition may be repeated at least 1 time, such as repeated at least 2 times, e.g. at least 3 times, such as repeated at least 5 times, e.g. at least 6 times, such as repeated at least 7 times, e.g. at least 8 times, such as repeated at least 9 times, e.g. at least 10 times.
  • step (iv)) may be repeated at least 3 times.
  • the encapsulated core material comprising an oil layer and surrounded by a lipid layer may be combined with the molten lipid composition
  • the lipid layer may provide an increased homogenous coating surround the core material (e.g. the core material comprising an oil layer) and increasing the protection of sensitive material, e.g. a microorganism, present in the core material
  • Particulate material comprising an encapsulated core material, wherein the encapsulated core material comprises an oil layer surrounded by a lipid layer.
  • the particulate material was prepared according to the following procedure:
  • Oil Red 0 is suspended in palm oil, coconut oil, canola oil, soybean oil, corn oil, rapeseed oil, cottonseed oil, olive oil, and sunflower oil, respectively.
  • Ball-head pins are placed into deep-well microtiter plate wells and 125 ul of each of the Oil Red 0 suspensions is pipetted.
  • the plate is placed inside -80-degrees freezer for at least 30 min.
  • lipid/modified lipid comprising stearyl alcohol modified with: 1% Lauric Acid or
  • the molten lipid is kept at a temperature that is slightly higher than its melting point.
  • the frozen Oil Red 0 suspensions are taken out of freezer and individual beads bound to ball-head pins are subjected to dipping into molten lipid for a short while followed by airdrying for several cycles (until the core Oil Red O-stained oil is completely covered by lipid) and the particulate material is provided.
  • the formed combination beads are monitored weekly on an ongoing basis to observe for peripheral migration of the core Oil Red 0 stain and/or vegetable oil.
  • All the particulate materials provided showed no peripheral migration of the core Oil Red 0 stain and/or vegetable oil after 6 months at room temperature.
  • a coating device was designed according to Figure 1 having an angle of the coating inlet of about 15° was used to produce the coated particles.
  • 10 oil mixtures to be coated were prepared by mixing 5 oil mixtures comprising sunflower oil and 5 oil mixtures comprising rapeseed oil mixed with a model active substance.
  • the model substance used in this example was the water-soluble dye Patent Blue.
  • a lipid coating agent was prepared by melting and mixing 99% stearyl alcohol and 1% myristic acid (w/w) at 80°C - about 25°C above the solidification temperature of the coating agent.
  • the oil mixture was fed to the material nozzle of the coating device providing a cloud of material to be coated when sprayed into the chamber of the coating device.
  • the coating agent was fed to the heated coating nozzles and sprayed into the coating device and forming a cloud of coating agent onto the oil mixture (material to be coated).
  • the coating agent solidifies on the core material in the chamber and the resulting coated particles was collected from the particulate material outlet.
  • the resulting coated particles had a mean particle size (D50) volume based of 41.7-88.0 pm.
  • the coating device according to the present invention showed to be highly useful for preparation of a particulate material comprising a core material surrounded by a coating, in particular on an industrial scale.
  • the coating device as used in Example 2 was used to produce coated particles with probiotics.
  • Freeze dried powder of mixed probiotic strains (Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium bifidum, Bifidobacterium braeve) was mixed with sunflower oil or and patent blue. The oil mixture was kept under stirring during the whole experiment. The probiotic content of the probiotic and oil was 5% on a weight basis.
  • Lipid coating material was prepared by melting and mixing 99% stearyl alcohol and 1% myristic acid (w/w) at 80°C and kept at temperature during the whole experiment.
  • the oil mixture (comprising the probiotics and the patent blue) was fed to the material nozzle providing a cloud of material to be coated when sprayed into the chamber of the coating device.
  • the coating agent was fed to the coating nozzles (heated to 80°C - about 25°C above the solidification temperature of the coating agent) and sprayed into the coating device and forming a cloud of coating agent onto the oil mixture (material to be coated).
  • the coating agent solidifies on the core material in the chamber and the resulting coated particles was collected from the particulate material outlet.
  • the resulting coated particles with probiotics had a mean particle size (D50) volume based of 56.2 pm, and viability of the probiotics was maintained high - above 90% viability after 14 days of encapsulation.
  • a microscopy image (polarized light microscopy) of the resulting coated particles of example 3, are shown in figure 3, with shell (lipid layer) visible as lighter part surrounding the darker core material and oil layer (core+oil are viewed together).
  • the coating device according to the present invention showed to be highly suitable for providing a particulate material comprising a core material containing probiotics surrounded by a coating where leakage of probiotics from the coated particles was not detectable and the survival rate of the probiotics was high.
  • coated particles produced in examples 2 and 3 were tested for retention, i.e., the capacity to retain the patent blue colorant and probiotics inside the coated particles when placed in water.
  • Coated particles were produced as described in Examples 2 and 3 with patent blue in the core oil or a combination of probiotics and patent blue.
  • Figure 2 The importance of the solidification process can be described by Figure 2. If the coating agent solidifies before reaching the surface of the material to be coated, or if the surface of the particles are not sufficiently in contact with the coating agent, the coating agent will not be homogenous and can result in a highly porous and partly aggregated structure (Figure 2A). If the coating is still liquid when reaching the surface of the material to be coated, and if sufficiently distributed over the surface of the material to be coated, the coating has the possibility to create a fully covering and homogenous layer before solidifying on the surface, which is preferred ( Figure 2B) which leads to a high retention of the particulate material.
  • Figure 2A If the coating is still liquid when reaching the surface of the material to be coated, and if sufficiently distributed over the surface of the material to be coated, the coating has the possibility to create a fully covering and homogenous layer before solidifying on the surface, which is preferred (Figure 2B) which leads to a high retention of the particulate material.

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Abstract

The present invention relates to a particulate material comprising an encapsulated core material, wherein the encapsulated core material comprises an oil layer, wherein the oil layer is surrounded by a lipid layer.

Description

ENCAPSULATED CORE MATERIAL
Technical field of the invention
The present invention relates to a method for providing a microencapsulated core material. In particular the present invention relates to a method of providing a microencapsulated core material, and the microencapsulated core material as such, which may safely and efficiently deliver probiotics and other sensitive particulate and non-particulate materials to target locations in the body.
Background of the invention
The human digestive tract comprises trillions of bacteria, many of which help digest food and fight off harmful bacteria. Studies have shown that some of these bacteria may influence a range of diseases, for better or worse.
It is found that manipulating these bacterial populations in the gastrointestinal tract, known collectively as the microbiome, could improve human health.
Varies strategies of manipulating the microbiome have been suggested and may be divided into two main groups, the prebiotic strategies and the probiotic strategies.
The prebiotic strategies relate to compounds in food that may induce the growth or activity of beneficial microorganisms, such as bacteria and fungi, in the gastrointestinal tract, where the prebiotics may be altering the composition of organisms in the microbiome.
Dietary prebiotics are typically nondigestible fibre compounds that pass undigested through the upper part of the gastrointestinal tract and stimulate the growth or activity of advantageous bacteria in the colon by acting as substrates for them.
The probiotic strategies relate to live microorganisms capable of or intended to provide health benefits when consumed. Generally, the health benefits may be provided by improving or restoring the gut microbiota of the mammal consuming the probiotic. Probiotics are considered generally safe to consume. One of the challenges of manipulating the bacterial population of beneficial bacteria in the intestine, in particular using probiotic, thus live microorganisms, may be to deliver large numbers of beneficial bacterial to the gut, where the microorganisms delivered must be live microorganisms, and therefore they need to survive the passage of the bile salts and acidic conditions of the stomach.
A number of microencapsulation techniques were being developed to address this problem and probiotics have been formulated into powders containing the bacteria to protect the live microorganisms (the probiotics). However, until now powdered probiotics taken orally are often destroyed or partly destroyed by the acidic conditions of the stomach, and low shelf life when formulated into the products, in particular into moisture product, despite the various microencapsulation techniques presently used and they cannot be incorporated into other products.
Thus, there is a need for an improved method for encapsulating sensitive materials, and improved encapsulated materials as such, which are more efficient; more reliable; which has an improved resistance against stomach conditions; has higher viability of encapsulated microorganisms, improved delivery of viable microorganisms to the microbiome of an animal or a human; which has higher stability during distribution and storage; and which has prolonged shelf life, in particular when formulated into products.
Summary of the invention
Thus, an object of the present invention relates to a method for providing a microencapsulated core material. In particular the present invention relates to a method of providing a microencapsulated core material, and the microencapsulated core material as such, which may safely and efficiently deliver probiotics and other sensitive particulate and non-particulate materials to target locations in the body, in particular the intestine.
In particular, it is an object of the present invention to provide an improved method of encapsulating sensitive materials and improved encapsulated materials, that solves the above-mentioned problems of the prior art with viability, stability, reliability and efficiency.
Thus, one aspect of the invention relates to a particulate material comprising an encapsulated core material, wherein the encapsulated core material comprises an oil layer, wherein the oil layer is surrounded by a lipid layer. Another aspect of the present invention relates to a method for providing a particulate material, the method comprises the steps of:
(i) Providing a core material;
(ii) Mixing the core material with an oil component, providing an encapsulated core material comprising an oil layer;
(iii) Providing a molten lipid composition;
(iv) Combining the encapsulated core material comprising an oil layer (from step (ii) with the molten lipid composition (from step (iii)), for providing a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
Yet another aspect of the present invention relates to a food product or a feed product comprising the particulate material according to the present invention.
Still another aspect of the present invention relates to a beverage comprising a particulate material according to the present invention.
A further aspect of the present invention relates to a medical product for use as a medicament.
An even further aspect of the present invention relates to a medical product for use in the treatment of gut diseases, immune diseases, skin diseases, and/or oral diseases.
Brief description of the figures
Figure 1 shows a coating device (1) according to the present invention. The coating device (1) a chamber (2), one material inlet (3), 3 coating inlet (4), and a particulate material outlet (5).
The material inlet (3) comprises at a material nozzle and each 3 coating inlets (4) comprises at least one coating nozzle (7) and each of the 3 coating nozzles (7) are angled relative to the material nozzle (6). This angle provides a spray zone (14) where the 3 clouds of coating agents are contacting the cloud of the material to be coated (and the particles in the cloud of the material to be coated) from different sides resulting in an improved coating of the particulate material and having a more complete coating of the particulate material.
The material inlet (3) may comprise a material feed (8) and a feed of compressed gas (9) and the 3 coating inlets (4) comprises a coating feed (10) and a feed of compressed gas (9).
The material inlet (3) is in fluid connection with a tank (11) comprising the material to be coated and the 3 coating inlets (4) are in fluid connection with a coating agent tank (12) comprising the coating agent.
To allow proper flowability the material inlet (3) and/or in particular the 3 coating inlets (4) are providing with means for heating.
The material nozzle (6) may provide a cloud of material to be coated with a density of particles which may be customized and the 3 coating nozzles (7) may provide clouds of coating agent with a density of particles which is customized, in such a way that coating of the individual particles may be provided.
The particulate material obtained from the particulate material outlet (5) may be send to a separation process using a device like a cyclone (13). The device may optionally be equipped with means for cooling.
Figure 2 shows various stages of solidification and the stage preferably reached by the present invention. The first stage (fig. 2a) shows the contact between a material to be coated and a coating material and how the coating agent is accumulated on the surface of the material to be coated, since the coating agent may be solidified before reaching the material to be coated.
In figure 2b it is shown the proper contact between a material to be coated and a coating material and how the coating agent stick to the surface of the material to be coated and spreads before being solidified. This is possible because the coating agent reaches the material to be coated before the coating agent is solidified.
Figure 3 shows a microscopy image (polarized light microscopy of coated particles according to the present invention, with a shell (lipid layer) visible as lighter part surrounding the darker core material provided with an oil layer.
The present invention will now be described in more detail in the following. Detailed description of the invention
The aim of the present invention relates to microencapsulation of sensitive materials, such as probiotics. To fill the gap with safe and efficient delivery of probiotics and other sensitive particulate and non-particulate materials to target locations in the body.
Accordingly, the inventor of the present invention found a method for providing a particulate material comprising an encapsulated core material.
In an embodiment of the present invention, the particulate material may be a powdered particulate material.
While the core material may comprise some water, in particular when the core material may be a microorganism, such as a lactic acid bacteria or a probiotic microorganism, the particulate material may have a moisture content below 5% (w/w), preferably below 3% (w/w), even more preferably below 1% (w/w), even more preferably no (or substantially no) moisture.
Microencapsulation techniques, or encapsulation, may be a process in the live microorganism may be surrounded by a coating to give small capsules.
The present invention relates to a novel microencapsulation to deliver live probiotics safely and efficiently to the ileal intestine and/or to the large intestine, preferably, to the large intestine.
Preferably, the present invention relates to a particulate material comprising an encapsulated core material including an oil-based true-core, preferably comprising one or more microorganism, and surrounded by a lipid-based true shell that may be highly tolerant to low gastric pH conditions and/or moisture.
The particulate material comprising an encapsulated core material may have a high- release rate in response to stimuli from the body when delivered to the microbiome in the gastrointestinal track.
A preferred embodiment of the present invention relates to a particulate material comprising an encapsulated core material, wherein the encapsulated core material may comprise an oil layer, wherein the oil layer may be surrounded by a lipid layer. The particulate material may be a small, often very small particles which may comprise a cores material.
In an embodiment of the present invention the particulate material may have a mean particle size (D50) less than 1000pm, such as less than 750pm, e.g. less than 500pm, such as less than 300pm, e.g. less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
In an embodiment of the present invention the particulate material may have a mean particle size (D90) less than 1000pm, such as less than 750pm, e.g. less than 500pm, such as less than 300pm, e.g. less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
The particulate material may vary greatly in size, shape, dispersible, and digestibility.
The core material according to the present invention may comprise a sensitive material. The sensitive material may be sensitive to the influence of chemicals, enzymes, oxygen, light, or a combination hereof.
In an embodiment of the present invention the core material may be a microorganism, food ingredients, cosmetic ingredients, pharmaceutical component, enzymes, dyes, vitamins, cells or other materials.
Preferably, the core material may not be an oil, such as a fish oil or a plant oil.
In an embodiment of the present invention the core material may comprise a microorganism.
Preferably, the sensitive material may be a microorganism.
Preferably, the microorganism may be a bacterium, a fungus, a yeast, or a virus.
In a preferred embodiment of the present invention the microorganism may be a viable microorganism. The viable microorganisms may be live microorganisms and/or viable spores.
In an embodiment of the present invention the core material may comprise one or more probiotic microorganism.
The probiotic microorganism may relate to live microorganisms which when administered in adequate amounts to a human or animal may confer a health benefit.
Preferably, the microorganism may be a microorganism which is capable of introducing, supporting or boosting the development of a beneficial gut microflora.
In an embodiment of the present invention the core material may comprise one or more bacterial strain.
Preferably, the one or more bacterial strain may include one or more lactic acid bacterial strains. Preferably, the one or more bacterial strain may consist essentially of one or more lactic acid bacterial strains.
Preferably, the one or more lactic acid bacterial strains may comprise Lactobacillus, Bifidobacterium or a combination hereof.
The core material may comprise a concentration of live or viable microorganism, in particular live or viable bacterial, in the range of 103-1015 CFU/g, such as in the range of 105-1014 CFU/g, e.g. in the range of 106-1012 CFU/g, such as in the range of IO7- 1011 CFU/g, e.g. in the range of 108-1010 CFU/g.
The term "CFU" relates to colony-forming unit and may be a unit which estimates the number of microbial cells (in particular bacteria) in a sample that are viable and thus able to multiply under controlled conditions.
The microorganisms provided for the core material may be provided in moisten state, in dry state or in frozen state before suspended in the component.
Preferably, the lipid layer and the oil layer according to the present invention may both comprises fatty acids. One difference between the lipid layer and the oil layer according to the present invention may be that at room temperature the lipid layer may be solid whereas oil layer may be liquid.
Preferably, room temperature according to the present invention, may be a temperature in the range of 18-25°C.
In an embodiment of the present invention the oil layer may comprise an oil component.
The oil component may comprise a single oil component of a combination of two or more oils compounds or a combination of oil and lipid compounds.
Preferably, the oil component may comprise a vegetable oil.
Preferably, the oil component may be liquid at a temperature below 30°C, e.g. the oil component may be liquid at a temperature below 25°C, e.g. the oil component may be liquid at a temperature below 20°C, e.g. the oil component may be liquid at a temperature below 15°C, e.g. oil component may be liquid at a temperature below 10°C, such as at a temperature in the range of 5-35°C, e.g. in the range of 10-30°C, such as in the range of 15-25°C, e.g. of about 25°C.
In an embodiment of the present invention the oil component and/or the oil layer may comprise palm oil, coconut oil, canola oil, soybean oil, corn oil, rapeseed oil, cottonseed oil, olive oil, sunflower oil or a combination hereof. Preferably, the oil component and/or the oil layer may comprise coconut oil, rapeseed oil, sunflower oil, or a combination hereof.
The core material may be encapsulated by the oil layer by mixing the core material according to the present invention with the oil component creating a core material comprising an oil layer.
Preferably, the oil layer may provide a coherent layer, preferably a coherent thin layer, of oil component surrounding the core material.
The oil layer may surround the core material as a hole uniform element, and/or the oil layer may surround individual parts of the core material making up the hole uniform element.
The lipid layer, preferably the coherent layer, may provide at least 20% encapsulation efficiency, such as at least 30% encapsulation efficiency, e.g. at least 40% encapsulation efficiency, such as at least 50% encapsulation efficiency, e.g. at least 60% encapsulation efficiency, such as at least 80% encapsulation efficiency, e.g. at least 90% encapsulation efficiency, such as at least 95% encapsulation efficiency, e.g. at least 98% encapsulation efficiency.
The term "encapsulation efficiency" may relate to the amount of core material maintained in the particulate material when exposed to water. The exposure to water may be washing with water. The washing with water may include a vigorously mixing and/or stirring.
The mean particle size (D50) of the core material surrounded by the oil layer may be less than 100 pm, such as less than 75 pm, e.g. less than 60 pm, such as less than 50 pm, e.g. less than 40 pm, such as less than 30 pm, e.g. less than 20 pm, such as less than 10 pm, e.g. in the range of 5-125 pm, such as in the range of 10-100 pm, e.g. in the range of 20-75 pm, such as in the range of 30-60 pm, e.g. in the range of 40-50 pm.
The mean particle size (D90) of the core material surrounded by the oil layer may be less than 100 pm, such as less than 75 pm, e.g. less than 60 pm, such as less than 50 pm, e.g. less than 40 pm, such as less than 30 pm, e.g. less than 20 pm, such as less than 10 pm, e.g. in the range of 5-125 pm, such as in the range of 10-100 pm, e.g. in the range of 20-75 pm, such as in the range of 30-60 pm, e.g. in the range of 40-50 pm.
The core material may be suspended in the oil component (or combination of oil components) to a substantially homogenous mixture.
In an embodiment of the present invention the core material may be suspended in the oil component (or combination of oil components) to provide a substantially homogenous mixture at a temperature below 30°C, e.g. the oil component may be liquid at a temperature below 25°C, e.g. the oil component may be liquid at a temperature below 20°C, e.g. the oil component may be liquid at a temperature below 15°C, e.g. oil component may be liquid at a temperature below 10°C, such as at a temperature in the range of 5-35°C, e.g. in the range of 10-30°C, such as in the range of 15-25°C, e.g. of about 25°C.
Preferably, the core material according to the present invention may constitute in the range of 1-50% (w/w) of the encapsulated core material comprises an oil layer, such as in the range of 10-45% (w/w), e.g. in the range of 15-40% (w/w), such as in the range of 20-35% (w/w), e.g. in the range of 25-30% (w/w). In an embodiment of the present invention the encapsulated core material comprising the oil layer may be surrounded by a lipid layer.
The lipid layer may be provided by a lipid compound. The lipid compound may comprise a single lipid compound, a combination of different lipid compounds, a combination of a single lipid compound and an oil, or a combination of two or more different lipid compounds and an oil.
In an embodiment of the present invention the lipid compound may a solid at a temperature below 25°C, e.g. the oil component may be a solid at a temperature below 20°C, e.g. the oil component may be a solid at a temperature below 15°C, e.g. oil component may be a solid at a temperature below 10°C, such as at a temperature in the range of 5-35°C, e.g. in the range of 10-30°C, such as in the range of 15-25°C, e.g. of about 25°C.
In another embodiment of the present invention the lipid compound may be liquid at a temperature below 70°C, e.g. the oil component may be liquid at a temperature below 65°C, e.g. the oil component may be liquid at a temperature below 60°C, e.g. the oil component may be liquid at a temperature below 55°C, e.g. the oil component may be liquid at a temperature below 50°C, e.g. the oil component may be liquid at a temperature below 40°C, e.g. the oil component may be liquid at a temperature below 30°C, e.g. the oil component may be liquid at a temperature below 20°C, such as at a temperature in the range of 20-70°C, e.g. in the range of 40-65°C, such as in the range of 50-60°C, e.g. in the range of 52-58°C.
In the present context the term "solid" relates to a compound that has a three-dimensional stable structure or shape and does not deform by the action of gravity.
In an embodiment of the present invention the lipid layer may comprise a fatty acid, a fatty alcohol, a wax, a sterol, a phosphor lipid, or a combination hereof. Preferably, the lipid layer may comprise a fatty acid, a fatty alcohol, or a combination hereof.
The fatty acid or fatty alcohol may preferably be a saturated fatty alcohol or a saturated fatty acid and/or the fatty acid or fatty alcohol may comprise a carbon chain length ranging from C12-C22, such as C12-C20, e.g. C12-C18.
Preferably, the lipid layer may comprise the combination of a fatty acid and a fatty alcohol. The presence of fatty alcohol in the lipid layer may be preferred over the presence and/or concentration of fatty acid.
In an embodiment of the present invention the fatty acid may be selected from lauric acid, myristic acid, or a combination hereof.
In another embodiment of the present invention the fatty alcohol may be selected from cetyl alcohol, stearyl alcohol, tetradecanol, or a combination hereof.
Preferably, the lipid layer may comprise stearyl alcohol in combination with lauric acid, myristic acid, palmitic acid or a combination hereof.
In a lipid layer comprising stearyl alcohol in combination with lauric acid, myristic acid, or a combination of lauric acid and myristic acid, the content of stearyl alcohol may represent at least 20% (w/w) of the lipid layer, such as at least 30% (w/w) of the lipid layer, e.g. at least 40% (w/w) of the lipid layer, such as at least 50% (w/w) of the lipid layer, e.g. at least 60% (w/w) of the lipid layer, such as at least 70% (w/w) of the lipid layer, e.g. at least 80% (w/w) of the lipid layer, such as at least 90% (w/w) of the lipid layer, e.g. at least 95% (w/w) of the lipid layer, such as about 98% (w/w) of the lipid layer, e.g. about
98.5% (w/w) of the lipid layer.
In a lipid layer comprising stearyl alcohol in combination with lauric acid, myristic acid, or a combination of lauric acid and myristic acid, the content of lauric acid may represent in the range of 0.1-10% (w/w), such as in the range of 0.2-5% (w/w), e.g. in the range of 0.3- 2% (w/w), such as in the range of 0.5-1% (w/w).
In a lipid layer comprising stearyl alcohol in combination with lauric acid, myristic acid, or a combination of lauric acid and myristic acid, the content of myristic acid may represent in the range of 0.1-10% (w/w), such as in the range of 0.2-5% (w/w), e.g. in the range of 0.3-2% (w/w), such as in the range of 0.5-1% (w/w).
The temperature of the lipid composition when applied to the encapsulated core material comprising an oil layer may be above the melting point of the lipid composition.
The melting point of the lipid composition may be the temperature and/or energy level necessary for changing the state of the lipid composition from a solid state to a liquid state
The melting point of the lipid composition may be dependent on the composition of the lipid composition. In an embodiment of the present invention, the temperature of the lipid composition, before contacting the lipid composition with the encapsulated core material comprising an oil layer, may be set in such a way that the lipid composition may be liquid at the time of contact with the encapsulated core material comprising an oil layer may be.
Following contact between the lipid composition and the encapsulated core material comprising an oil layer, the lipid composition may be surrounding the encapsulated core material comprising an oil layer and the lipid layer may be allowed to solidify, creating a lipid layer. Thus, a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer, may be provided.
The term "solidify" means that the lipid composition (or the lipid layer) changes state from a liquid state to a solid state.
In an embodiment of the present invention the temperature of the lipid composition at the time of contacting the encapsulated core material comprising an oil layer may be above the melting point of the lipid composition. Preferably, the lipid composition has a temperature above 25°C, such as above 30°C, e.g. above 35°C, such as above 40°C, e.g. above 45°C, such as above 50°C, e.g. above 55°C, such as above 60°C, e.g. above 65°C, such as in the range of 25-70°C, e.g. in the range of 30-65°C, such as in the range of 35- 60°C, e.g. in the range of 40-55°C, such as in the range of 45-50°C.
Preferably, the temperature of the lipid composition may be set at a temperature just slightly above the melting point of the lipid composition. In an embodiment of the present invention the temperature of the lipid composition may be set at a temperature in the range of l-20°C above the melting point of the lipid composition, such as in the range of 1-15°C above the melting point of the lipid composition, e.g. in the range of 2-10°C above the melting point of the lipid composition, such as in the range of 3-8°C above the melting point of the lipid composition, e.g. in the range of 4-6°C above the melting point of the lipid composition.
Following the addition of the lipid layer to the encapsulated core material comprising an oil layer, the encapsulated core material comprising an oil layer and surrounded by a lipid layer may be subjected to cooling in order to solidify the lipid layer around the encapsulated core material comprising an oil layer providing the particulate material according to the present invention. The particulate material according to the present invention may maintain at least 10% of the core material, such as at least 20% of the core material, e.g. at least 30% of the core material, such as at least 40% of the core material, e.g. at least 50% of the core material, such as at least 60% of the core material, e.g. at least 70% of the core material, such as at least 80% of the core material, e.g. at least 90% of the core material, encapsulated in the oil layer and the lipid layer after a period of at least 10 minutes in a test solution, such as for a period of at least 30 minutes, e.g. for at least 45 minutes, such as for a period of at least 60 minutes, e.g. for at least 2 hours, such as for a period of at least 6 hours, e.g. for at least 12 hours, such as for a period of at least 24 hours, e.g. for at least 2 days, such as for a period of at least 5 days, e.g. for at least 10 days, such as for a period of at least 20 days, e.g. for at least 30 days, such as for a period of at least 40 days, e.g. for at least 50 days, such as for a period of at least 60 days, e.g. for at least 70 days.
The particulate material may comprise a core material comprising (consisting essentially of) viable or live microorganisms, in particular one or more probiotics, and at least 10% of the microorganisms, in particular the probiotics, such as at least 20%, e.g. at least 30%, such as at least 50%, e.g. at least 60%, such as at least 70%, e.g. at least 80% of the core material, such as at least 90% of the core material, e.g. at least 95%, of the microorganisms, in particular one or more probiotics, remains alive or viable, after storage for a period of at least 10 minutes in a test solution, such as for a period of at least 30 minutes, e.g. for at least 45 minutes, such as for a period of at least 60 minutes, e.g. for at least 2 hours, such as for a period of at least 6 hours, e.g. for at least 12 hours, such as for a period of at least 24 hours, e.g. for at least 2 days, such as for a period of at least 5 days, e.g. for at least 10 days, such as for a period of at least 20 days, e.g. for at least 30 days, such as for a period of at least 40 days, e.g. for at least 50 days, such as for a period of at least 60 days, e.g. for at least 70 days.
In an embodiment of the present invention the core material comprises one or more viable microorganisms and wherein one or more microorganisms show a reduction in viability of at most 3 LOG over a period of storage of 12 months at room temperature.
In a further embodiment of the present invention storage of the particulate material comprising a core material comprising (consisting essentially of) one or more viable microorganisms according to the present invention and wherein the one or more viable microorganisms show a reduction in viability of at most 3 LOG, such as a reduction of at the most 2 LOG, e.g. a reduction of at the most 1 LOG reduction, such as a reduction of at the most 0.5 LOG, e.g. a reduction of at the most 0.1 LOG reduction over a period of storage of 12 months at room temperature. The test solution may be selected from water, aqueous acidic environment, or an aqueous environment comprising bile salts and or bile acids.
Preferably, the surface of the encapsulated core material comprising an oil layer of the particulate material may be covered or partly covered by the lipid layer. In an embodiment of the present invention the encapsulated core material comprising an oil layer of the particulate material may be at least 40% covered by the lipid layer, such as at least 50% covered, e.g. at least 60% covered, such as at least 70% covered, e.g. at least 80% covered, such as at least 85% covered, e.g. at least 90% covered, such as at least 95% covered.
The effect of the lipid layer may be determined by the efficiency to retain a core material in the particulate material. The larger part of the particulate material being covered by the lipid layer (i.e. the higher the coverage), the higher the amount of the core material may be retained in the particle material and the less susceptible the particulate material is to obtain material from the outside of the particulate material.
The particulate material according to the present invention may provide a retention of a trace element of 70% or above, such as 75% or above, e.g. 80% or above, such as 85% or above, e.g. 90% or above, such as 92% or above, e.g. 94% or above, such as 96% or above, e.g. 98% or above.
Preferably, the particulate material according to the present invention may provide a retention of the core material of 70% or above, such as 75% or above, e.g. 80% or above, such as 85% or above, e.g. 90% or above, such as 92% or above, e.g. 94% or above, such as 96% or above, e.g. 98% or above.
In an embodiment of the present invention, the encapsulation efficiency and/or the lipid coverage or the retention properties of the particulate material may be determined by a diffusion/retention test using a trace element.
The trace element may be Patent Blue Colorant, Oil Red O or the like.
The outer surface of a particulate material comprising a microbial core material, according to the present invention, may comprise less than 106 CFU/g particulate material, such as less than 105 CFU/g particulate material, e.g. less than 104 CFU/g particulate material, such as less than 103 CFU/g particulate material, e.g. less than 102 CFU/g particulate material. A preferred embodiment of the present invention relates to a method for providing a particulate material, the method comprises the steps of:
(i) Providing a core material;
(ii) Mixing the core material with an oil component, providing an encapsulated core material comprising an oil layer;
(iii) Providing a molten lipid composition;
(iv) Combining the encapsulated core material comprising an oil layer (from step (ii) with the molten lipid composition (from step (iii)), providing a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
Preferably, agitation may be provided during mixing in step (ii).
The combination of the encapsulated core material comprising and an oil layer with the molten lipid composition (step (iv)) may be performed in a coating device, the coating device comprising a chamber, wherein the chamber comprising at least one material inlet and at least one coating inlet and a particulate material outlet.
The at least one material inlet and/or the at least one coating inlet may be placed in the upper part of the chamber.
In an embodiment of the present invention each of the at least one material inlet may comprise at least one material nozzle and/or wherein each the at least one coating inlet comprises at least one coating nozzle.
Preferably, the at least one coating nozzle may be angled relative to the material nozzle.
In an embodiment of the present invention the material nozzle of the material inlet may be pointing vertically downwards, or substantially vertically downwards.
By pointing the material nozzle of the material inlet vertically downwards, or substantially vertically downwards, it may be easier to control the cloud created of the material to be coated and/or limiting the risk of having the cloud provided to collapse. The coating nozzle may be angled by 1-179° relative to the material nozzle, such as by 2- 150°, e.g. by 3-125°, such as by 4-100°, e.g. by 5-75°, such as by 6-50°, e.g. by 7-35°, such as by 8-25°, e.g. by 9-20°, such as by 10-18°, e.g. about 15° .
In an embodiment of the present invention the at least one material inlet and/or the at least one coating inlet are providing with means for heating.
The at least one material inlet may be in fluid connection with a tank comprising the material to be coated.
The at least one coating inlet may be in fluid connection with a coating agent tank comprising the coating agent.
Preferably, the chamber may comprise at least two coating inlets, such as at least 3 coating inlets, e.g. at least 4 coating inlets, such as at least 5 coating inlets, e.g. at least 6 coating inlets, such as at least 7 coating inlets, e.g. at least 8 coating inlets.
In an embodiment of the present invention the chamber comprising of the coating device is provided with one material inlet and at least one coating inlet, e.g. at least two coating inlets, such as at least 3 coating inlets, e.g. at least 4 coating inlets, such as at least 5 coating inlets, e.g. at least 6 coating inlets, such as at least 7 coating inlets, e.g. at least 8 coating inlets.
Preferably, the two or more coating inlets are surrounding the material inlet to improve the distribution of the lipid layer on the surface of the encapsulated core material comprising and an oil layer, and at the same time keeping the mean particle size low.
The method of coating the encapsulated core material comprising and an oil layer with the lipid layer may comprise the steps of:
(i) Injecting the encapsulated core material comprising and an oil layer into the coating device as described herein providing a cloud of the material to be coated inside a chamber of the coating device;
(ii) Injecting the lipid composition into the coating device providing a cloud of the lipid composition inside a chamber of the coating device, wherein the cloud of the lipid composition may be injected into the cloud of the encapsulated core material comprising and an oil layer to be coated, providing the particulate material.
In an embodiment of the present invention the combining of the encapsulated core material comprising an oil layer (from step (ii) with the molten lipid composition (from step (Hi)), providing a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
Preferably, the combining may involve putting or applying the molten lipid composition on the top or the surface of the encapsulated core material comprising an oil layer, resulting in the particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
In an embodiment of the present invention the particles of the encapsulated core material comprising an oil layer and a lipid layer may comprise a mean particle size (D50) of less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
In an embodiment of the present invention the particles of the encapsulated core material comprising an oil layer and a lipid layer may comprise a mean particle size (D90) of less than 150 pm, such as less than 125 pm, e.g. less than 100 pm, such as less than 75 pm, e.g. less than 50 pm, such as less than 40 pm, e.g. less than 30 pm, such as in the range of 30-150 pm, e.g. in the range of 40-125 pm, such as in the range of 50-100 pm, e.g. in the range of 60-75 pm.
The mean particle size or the average particles size may be determined by sieving, sedimentation electrozone testing, automated microscopy and/or by laser diffraction.
The mean particle size may be selected according to the use or application of the particulate material, or the encapsulated core material comprising an oil layer and surrounded by a lipid layer.
The lipid layer may be allowed to solidify, e.g. by subjecting the encapsulated core material comprising an oil layer and surrounded to e.g. cooling, providing a particulate material according to the present invention. The particulate material according to the present invention may be a substantially spherical particulate material.
An embodiment of the present invention the encapsulated core material comprising an oil layer and surrounded by a lipid layer (obtained in step (iv)) may be combined with the molten lipid composition.
As mentioned the encapsulated core material comprising an oil layer and surrounded by a lipid layer (obtained in step (iv)) may be combined with the molten lipid composition to increase the thickness of the lipid layer of the encapsulated core material comprising an oil layer and surrounded by a lipid layer.
In an embodiment of the step of combining the encapsulated core material comprising an oil layer and surrounded by a lipid layer with the molten lipid composition may be repeated at least 1 time, such as repeated at least 2 times, e.g. at least 3 times, such as repeated at least 5 times, e.g. at least 6 times, such as repeated at least 7 times, e.g. at least 8 times, such as repeated at least 9 times, e.g. at least 10 times.
In a further embodiment of the present invention the step of combining the encapsulated core material comprising an oil layer and surrounded by a lipid layer (obtained in step (iv)) may be repeated at least 3 times.
By repeating the number of circles the encapsulated core material comprising an oil layer and surrounded by a lipid layer (obtained in step (iv)) may be combined with the molten lipid composition, the lipid layer may provide an increased homogenous coating surround the core material (e.g. the core material comprising an oil layer) and increasing the protection of sensitive material, e.g. a microorganism, present in the core material
It should be noted that embodiments and features described in the context of one of the aspects of the present invention also apply to the other aspects of the invention.
All patent and non-patent references cited in the present application, are hereby incorporated by reference in their entirety.
The invention will now be described in further details in the following non-limiting examples. Examples
Example 1
Method of producing the particulate material according to the present invention. Particulate material comprising an encapsulated core material, wherein the encapsulated core material comprises an oil layer surrounded by a lipid layer.
The particulate material was prepared according to the following procedure:
9 solutions were prepared comprising 1% (w/v) Oil Red 0 is suspended in palm oil, coconut oil, canola oil, soybean oil, corn oil, rapeseed oil, cottonseed oil, olive oil, and sunflower oil, respectively.
Ball-head pins are placed into deep-well microtiter plate wells and 125 ul of each of the Oil Red 0 suspensions is pipetted.
The plate is placed inside -80-degrees freezer for at least 30 min.
In the meantime, lipid/modified lipid comprising stearyl alcohol modified with: 1% Lauric Acid or
0.5 Lauric Acid and 1% Myristic Acid is melted by putting the lipid in a glass bottle in a both of boiling water.
The molten lipid is kept at a temperature that is slightly higher than its melting point.
The frozen Oil Red 0 suspensions are taken out of freezer and individual beads bound to ball-head pins are subjected to dipping into molten lipid for a short while followed by airdrying for several cycles (until the core Oil Red O-stained oil is completely covered by lipid) and the particulate material is provided.
The formed combination beads are monitored weekly on an ongoing basis to observe for peripheral migration of the core Oil Red 0 stain and/or vegetable oil.
All the particulate materials provided showed no peripheral migration of the core Oil Red 0 stain and/or vegetable oil after 6 months at room temperature.
Example 2
A coating device was designed according to Figure 1 having an angle of the coating inlet of about 15° was used to produce the coated particles. 10 oil mixtures to be coated were prepared by mixing 5 oil mixtures comprising sunflower oil and 5 oil mixtures comprising rapeseed oil mixed with a model active substance. The model substance used in this example was the water-soluble dye Patent Blue.
A lipid coating agent was prepared by melting and mixing 99% stearyl alcohol and 1% myristic acid (w/w) at 80°C - about 25°C above the solidification temperature of the coating agent.
The oil mixture was fed to the material nozzle of the coating device providing a cloud of material to be coated when sprayed into the chamber of the coating device.
The coating agent was fed to the heated coating nozzles and sprayed into the coating device and forming a cloud of coating agent onto the oil mixture (material to be coated).
The coating agent solidifies on the core material in the chamber and the resulting coated particles was collected from the particulate material outlet.
The resulting coated particles had a mean particle size (D50) volume based of 41.7-88.0 pm.
Conclusion from Example 2:
The coating device according to the present invention showed to be highly useful for preparation of a particulate material comprising a core material surrounded by a coating, in particular on an industrial scale.
Example 3
The coating device as used in Example 2 was used to produce coated particles with probiotics.
Freeze dried powder of mixed probiotic strains (Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium bifidum, Bifidobacterium braeve) was mixed with sunflower oil or and patent blue. The oil mixture was kept under stirring during the whole experiment. The probiotic content of the probiotic and oil was 5% on a weight basis.
Lipid coating material was prepared by melting and mixing 99% stearyl alcohol and 1% myristic acid (w/w) at 80°C and kept at temperature during the whole experiment. The oil mixture (comprising the probiotics and the patent blue) was fed to the material nozzle providing a cloud of material to be coated when sprayed into the chamber of the coating device.
The coating agent was fed to the coating nozzles (heated to 80°C - about 25°C above the solidification temperature of the coating agent) and sprayed into the coating device and forming a cloud of coating agent onto the oil mixture (material to be coated).
The coating agent solidifies on the core material in the chamber and the resulting coated particles was collected from the particulate material outlet.
The resulting coated particles with probiotics had a mean particle size (D50) volume based of 56.2 pm, and viability of the probiotics was maintained high - above 90% viability after 14 days of encapsulation. A microscopy image (polarized light microscopy) of the resulting coated particles of example 3, are shown in figure 3, with shell (lipid layer) visible as lighter part surrounding the darker core material and oil layer (core+oil are viewed together).
Conclusion from Example 3:
The coating device according to the present invention showed to be highly suitable for providing a particulate material comprising a core material containing probiotics surrounded by a coating where leakage of probiotics from the coated particles was not detectable and the survival rate of the probiotics was high.
Example 4
Testing retention of the coated particles produced in examples 2 and 3.
The coated particles produced in examples 2 and 3 were tested for retention, i.e., the capacity to retain the patent blue colorant and probiotics inside the coated particles when placed in water.
Coated particles were produced as described in Examples 2 and 3 with patent blue in the core oil or a combination of probiotics and patent blue.
Process adjustments affecting the solidification of the lipid coating are noted with the results below. The retention was measured by the following procedure. 50 mL cold demineralized water was added to a beaker. For comparison of full release of colorant, boiling demineralized water was added to another beaker. Both beakers were stirred at 500 rpm. 2 g powder was added simultaneously to each beaker and stirred for 2 min. After 2 min the stirring was stopped, and the samples left for additional 30 min, before taking a sample of each with a syringe and passing the sample through a 0.45 pm PVDF filter. The content of Patent Blue released from the particulate material was determined by reading the absorbance at 637 nm. The content of probiotics released from the particulate material was determined by plate spreading and colony counting. The retention was calculated as % patent blue released or % probiotics released to the cold water as compared to the full release (hot water sample).
With the process using the coating device according to the present invention a retention in the range of 89.9-96% was obtained.
Compared to conventional methods of producing coated particles below 250 pm, such conventionally produced particles show less than 70% retention, which is significantly less than what may be achieved by the device according to the present invention.
Conclusion from Example 4:
Proper solidification of the coating agent is important to produce coated particles with high retention - forming a "true core".
The importance of the solidification process can be described by Figure 2. If the coating agent solidifies before reaching the surface of the material to be coated, or if the surface of the particles are not sufficiently in contact with the coating agent, the coating agent will not be homogenous and can result in a highly porous and partly aggregated structure (Figure 2A). If the coating is still liquid when reaching the surface of the material to be coated, and if sufficiently distributed over the surface of the material to be coated, the coating has the possibility to create a fully covering and homogenous layer before solidifying on the surface, which is preferred (Figure 2B) which leads to a high retention of the particulate material.

Claims

Claims
1. A particulate material comprising an encapsulated core material, wherein the encapsulated core material is provided with an oil layer, and wherein the oil layer is surrounded by a lipid layer, wherein the encapsulated core material comprising an oil layer of the particulate material comprises at least 40% covered by the lipid layer
2. The particulate material according to claim 1, wherein the particulate material comprises a retention of a trace element or a core material above 70%.
3. The particulate material according to anyone of claims 1-2, wherein the particulate material has a mean particle size less than 125 pm.
4. The particulate material according to anyone of claims 1-3, wherein the core material comprises a microorganism, a food ingredient, a cosmetic ingredient, a pharmaceutical component, an enzyme, a dye, a vitamin or a combination hereof.
5. The particulate material according to claim 4, wherein the microorganism is a viable microorganism.
6. The particulate material according to anyone of claims 1-5, wherein the lipid layer comprises a fatty acid, a fatty alcohol, or a combination hereof.
7. The particulate material according to claim 6, wherein the fatty acid is selected from myristic acid, lauric acid, palmitic acid, or a combination hereof and wherein the fatty alcohol is selected from stearyl alcohol, cetyl alcohol, tetradecanol, or a combination hereof.
8. The particulate material according to anyone of claims 6-7, wherein the lipid layer comprises lauric acid, myristic acid, cetyl alcohol, stearyl alcohol, or any combination hereof.
9. The particulate material according to anyone of claims 1-8, wherein the oil layer comprises plant oil, such as palm oil, coconut oil, canola oil, soybean oil, corn oil, rapeseed oil, cottonseed oil, olive oil, sunflower oil or a combination hereof.
10. The particulate material according to anyone of claims 1-9, wherein the core material comprises one or more viable microorganisms and wherein one or more microorganisms show a reduction in viability of at most 3 LOG over a period of storage of 12 months at room temperature (a temperature in the range of 18-25°C).
11. The particulate material according to anyone of the preceding claims, wherein the particulate material may have a moisture content below 3% (w/w), preferably no (or substantially no) moisture.
12. The particulate material according to anyone of the preceding claims, wherein the encapsulated core material comprising an oil layer of the particulate material comprises at least 50% covered, e.g. at least 60% covered, such as at least 70% covered, e.g. at least 80% covered, such as at least 85% covered, e.g. at least 90% covered, such as at least 95% covered.
13. The particulate material according to anyone of the preceding claims, wherein the particulate material is providing a retention of a trace element or a core material of 75% or above, e.g. 80% or above, such as 85% or above, e.g. 90% or above, such as 92% or above, e.g. 94% or above, such as 96% or above, e.g. 98% or above.
14. The particulate material according to anyone of the preceding claims, wherein the core material is not an oil, such as a fish oil or a plant oil.
15. A method for providing a particulate material, the method comprises the steps of:
(i) Providing a core material;
(ii) Mixing the core material with an oil component, providing an encapsulated core material comprising an oil layer;
(iii) Providing a molten lipid composition;
(iv) combining the encapsulated core material comprising an oil layer (from step (ii) with the molten lipid composition (from step (iii)), for providing a particulate material comprising an encapsulated core material comprising an oil layer and surrounded by a lipid layer.
16. The method according to claim 15, wherein the combination of the encapsulated core material comprising and an oil layer with the molten lipid composition (step (iv) may be performed in a coating device, the coating device comprising a chamber (2), wherein the chamber (2) comprising at least one material inlet (3) and at least one coating inlet (4) and a particulate material outlet (5)..
PCT/EP2023/083775 2022-12-01 2023-11-30 Encapsulated core material Ceased WO2024115675A1 (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019191325A1 (en) * 2018-03-27 2019-10-03 Gpcp Ip Holdings Llc Microencapsulated probiotic and low-water-activity compositions containing the same
US20220265527A1 (en) * 2019-09-24 2022-08-25 Gpcp Ip Holdings Llc Microencapsulated probiotic and compositions containing the same

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019191325A1 (en) * 2018-03-27 2019-10-03 Gpcp Ip Holdings Llc Microencapsulated probiotic and low-water-activity compositions containing the same
US20220265527A1 (en) * 2019-09-24 2022-08-25 Gpcp Ip Holdings Llc Microencapsulated probiotic and compositions containing the same

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