TSEKOU CHRISTOS (CH)
URBAN KAI (CH)
SCHAEFER CHRISTIAN (CH)
WO2017017248A1 | 2017-02-02 | |||
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WO2019185940A1 | 2019-10-03 | |||
WO2019185904A1 | 2019-10-03 | |||
WO2019185942A1 | 2019-10-03 | |||
WO2019185888A1 | 2019-10-03 | |||
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Claims 1. A milk replacer comprising an animal-free formulation comprising a) Vitamin A or a C1-20 alkyl ester thereof and optionally vitamin D; b) Gum Arabic in an amount of ≤ 25 weight; c) at least one ingredient selected from modified food starches, starch hydrolysates, mono- and disaccharides; d) at least an antioxidant; e) an anti-caking agent; f) optionally an oil; g) optionally residual moisture; whereby all amounts a) to f) sum up to 100 weight-% and are based on the sum of the weight of a) to f), whereby the compound a) is encapsulated in a matrix formed by compounds b) and c). 2. The milk replacer according to claim 1, wherein the milk replacer is used for young non-human mammals, preferably for young non-human mammals selected from calves, foals, young goats, young sheep and camel foals. 3. An animal-free formulation comprising a) Vitamin A or a C1-20 alkyl ester thereof and optionally vitamin D; b) Gum Arabic in an amount of ≤ 25 weight; c) at least one ingredient selected from modified food starches, starch hydrolysates, mono- and disaccharides; d) at least an antioxidant; e) an anti-caking agent; f) optionally an oil; g) optionally residual moisture; whereby all amounts a) to f) sum up to 100 weight-% and are based on the sum of the weight of the ingredients a) to f), whereby the compound a) is encapsulated in a matrix formed by compounds b) and c). 4. The animal-free formulation according to claim 3, whereby said formulation has a bulk density ranging from 0.4 to 0.9 g/cm3. 5. The animal-free formulation according to claim 3 and/or 4, whereby the amount of the ingredient(s) c) is ranging from 35 to 58 weight-%, preferably it is ranging from 38 to 55 weight-%, more preferably it is ranging from 40 to 52 weight-%, based on the sum of the weight of the ingredients a) to f). 6. The animal-free formulation according to any one or more of claims 3 to 5, wherein the ingredient c) is either a starch hydrolysate with a DE ≤ 40 or a mixture of a modified food starch and a disaccharide, whereby the weight ratio of the modified food starch to the disaccharide is preferably ranging from 1:2 to 2:1. 7. The animal-free formulation according to any one or more of claims 3 to 6, wherein said anti-caking agent e) has a particle size D(v,50%) ranging from 100 nm to 10 μm, measured as dry dispersion with a Malvern MasterSizer 3000 (laser diffraction). 8. The animal-free formulation according to any one or more of claims 3 to 7, wherein the amount of Vitamin A or a C1-20 alkyl ester thereof is ranging from 10 to 29 weight-%, preferably it is ranging from 12 to 24 weight-%, more preferably it is ranging from 15 to 23 weight-%, most preferably it is ranging from 17 to 22 weight-%, based on the sum of the weight of the ingredients a) to f). 9. The animal-free formulation according to any one or more of claims 3 to 8, wherein the amount of the Gum Arabic b) is ranging from 10 to 25 weight-%, preferably it is ranging from 15 to 25 weight-%, most preferably it is ranging from 19 to 24.5 weight-%, based on the sum of the weight of the ingredients a) to f). 10. The animal-free formulation according to any one or more of claims 3 to 9, wherein the amount of the antioxidant d) is ≤ 7.5 weight-%, preferably it is ranging from 0.5 to 7.5 weight-%, based on the sum of the weight of the ingredients a) to f). 11. The animal-free formulation according to any one or more of claims 3 to 10, wherein the amount of the anti-caking agent e) is ranging from 0.01 to 10 weight-%, based on the sum of the weight of the ingredients a) to f). 12. Use of the formulation according to any one or more of claims 3 to 11 in a milk replacer. 13. The use according to claim 12, wherein the milk replacer is used for calves, foals, young sheep, young goats and young camels. 14. A process for the manufacture of a formulation according to any one or more of claims 3 to 11 comprising the following steps: A) Dissolving Gum Arabic b), the ingredient(s) c) and, if present, a water- soluble antioxidant(s) d) in water to obtain a matrix; B) Heating the fat-soluble vitamin(s) a), the fat-soluble antioxidant(s) d) and, if present, the oil f) to obtain an active phase; C) Emulsifying the active phase obtained in step B) into the matrix obtained in step A) to obtain a dispersion; D) Drying the dispersion obtained in step C), optionally in presence of an anti-caking agent, to obtain the formulation. 15. A process for the manufacture of a milk replacer according to claim 1 and/or claim 2 by mixing an animal-free formulation according to any one or more of claims 3 to 11 with essential nutrients including proteins, amino acids, carbohydrates, fats, vitamins and minerals. |
wherein R 1a and R 2a are independently from each other H or C 1-11 -alkyl or (CH 2 ) n ─OH with n being an integer from 1 to 4, or R 1a and R 2a represent together a keto group, A is CHR 3a or C(=O), and wherein R 3a , R 4a and R 6a are independently from each other H or C 1-4 -alkyl, and wherein R 5a is H or OH or C 1-4 -alkyl or C 1-4 -alkoxy, as disclosed in WO 2019/185894. Further suitable tocopherols are compounds of formula (II), wherein one of the two substituents R 1a and R 2a is C 12-21 -alkyl and the other of the two substituents R 1a and R 2a is either hydrogen or C1-5-alkyl or (CH 2 )n-OH with n being an integer from 1 to 5, and wherein A is CH(R 3a ), and wherein R 3a , R 4a and R 6a are independently from each other H or C 1-4 -alkyl, and wherein R 5a is H or OH or C 1-4 -alkyl or C 1-4 -alkoxy, as disclosed in WO 2019/185938. Compounds of formula (II), wherein A is CH 2 , R 1a is C 1-5 -alkyl, R 2a is either H or C 1-2 - alkyl, R 5a is either H or C 1-4 -alkoxy or C 1-4 -alkyl, and R 4a and R 6a are independently from each other either H or C 1-4 -alkyl, with the preferences as disclosed in WO 2019/185900 are also suitable antioxidants in the formulations of the present invention. Preferred examples of the antioxidants of formula (II) as disclosed in WO 2019/185894 are the following compounds of formula (1)-(11) with “Me” being methyl: Further examples of suitable antioxidants that can be used in the formulations of the present invention are compounds of formula (III) and (IV), wherein R 1b and R 2b are independently from each other H or C 1-11 -alkyl or (CH 2 ) n ─OH with n being an integer from 1 to 6 or R 1b and R 2b together represent a keto group, and wherein R 3b , R 4b , R 5b , and R 6b are independently from each other H or C 1-6 -alkyl or C 1-6 -alkoxy, and R 7b is H or C 1-6 -alkyl, as disclosed in WO 2019/185898. “alkyl” and “alkoxy” hereby encompass linear alkyl and branched alkyl, and linear alkoxy and branched alkoxy, respectively. Preferred examples of compounds of formula (III) and (IV) are the following compounds (12)-(19):
Further suitable antioxidants are compounds of formula (V), whereby R 1 , R 2 and R 3 are independently from each other H or linear C 1-6 -alkyl or branched C 3-8 -alkyl, whereby preferably R 1 is H or methyl or ethyl or n-propyl or iso-propyl or tert-butyl and R 2 and R 3 are independently from each other H or methyl or ethyl, with the further preferences as disclosed in WO 2019/185940. Also, the compounds of formula (VI) with n being 1 or 2, R 1b and R 3b being independently from each other H or C 1-5 -alkyl, and R 2b being either H or C 1-5 -alkyl or C1-5-alkyloxy, preferably with the proviso at least one of R 1b , R 2b and R 3b being H, as disclosed in WO 2019/185904 can be used as antioxidants in the formulations of the present invention. Hereby the following compounds of formulae (VI-1) and (VI-2) are especially preferred: The asterisks * mark each a chiral/stereogenic center, i.e. all possible isomers having any configuration at said centers are encompassed by the term “compound of formula (VI-1)” and “compound of formula (VI-2)”, respectively. Also, suitable antioxidants are compounds of the following formulae (VII) and (VIII) with R 1c , R 2c and R 3c being independently from each other H or C 1-4 -alkyl as published in WO 2019/185942 and WO 2019/185888, respectively. Preferred examples thereof are tocotrienols and tocopherols of the formulae (20) to (27) as shown below.
The asterisks * mark each a chiral/stereogenic center. The term “compound of formula (VII)/(VIII)” encompasses all possible isomers having any configuration at said centers. Especially preferred examples of the compound of formula (VII) are the following compounds of formulae (20) (= alpha-tocotrienol), (21) (= beta-tocotrienol), (22) (= gamma-tocotrienol) and (23) (= delta-tocotrienol), whereby all possible diastereomers and enantiomers are included.
Especially preferred examples of the compound of formula (VIII) are the following compounds of formulae (20) (= alpha-tocopherol), (21) (= beta-tocopherol), (22) (= gamma-tocopherol) and (23) (= delta-tocopherol), whereby all possible diastereomers and enantiomers are included. The asterisks * mark each a chiral/stereogenic center. The term “compound of formula (20)/(21)/(22)/(23)/(24)/(25)/(26)/(27)” encompasses all possible isomers having any configuration at said centers. Polyphenols Examples of suitable polyphenols are 2,4,5-trihydroxybutyrophenone, epigallo- catechin gallate (“EGCG”), epigallo-catechin, gallo-catechin, hydroxytyrosol, resveratrol, carnosol, 2-(3,4-dihydroxyphenyl)acetic acid and C 1-6 alkyl esters thereof, and any mixture thereof. Further suitable polyphenols are derivatives, preferably esters and (earth) alkali metal salts, of hydroxybenzoic acids such as e.g. gallic acid (= 3,4,5-trihydroxybenzoic acid) and syringic acid (= 4-hydroxy-3,5-dimethoxy-benzoic acid). Examples of preferred esters are C1-20 alkyl esters of gallic acid such as e.g. propyl gallate, octyl gallate or dodecyl gallate, and C 1-20 alkyl esters of syringic acid. Also, derivatives, preferably esters and (earth) alkali metal salts, of cinnamic acid and hydroxycinnamic acids such as e.g. ferulic acid (= 3-(4-hydroxy-3- methoxyphenol)prop-2-enoic acid), caffeic acid (= 3,4-Dihydroxycinnamic acid), dihydrocaffeic acid (= 3-(3,4-dihydroxyphenyl) propanoic acid), chlorogenic acid (= the ester of caffeic acid and (−)-quinic acid), o-, m-, p-coumaric acid (= 2-/3-/4- hydroxycinnamic acid), rosmarinic acid (= a caffeic acid ester of 3-(3,4- dihydroxyphenyl)lactic acid), or sinap(in)ic acid (= 3,5-dimethoxy-4-hydroxycinnamic acid) may be used as antioxidants in the present invention. Examples of derivatives of cinnamic acid are Z-ethoxyethyl p-methoxycinnamate, ethylhexyl p-methoxycinnamate, 2-ethylhexyl 4-methoxycinnamate, methyl diiso- propylcinnamate, isoamyl 4—methoxycinnamate, and diethanolamin 4-methoxy- cinnamate. The most preferred antioxidants used in the formulation of the present invention are DL-alpha-tocopherol, as well as a mixture of DL-alpha-tocopherol and sodium ascorbate, a mixture of DL-alpha-tocopherol and ascorbic acid and a mixture of DL-alpha-tocopherol and ascorbyl palmitate. The weight ratio of DL-alpha-tocopherol to sodium ascorbate and of DL-alpha- tocopherol to ascorbic acid is preferably ranging from 5:1 to 1:5, more preferably from 2:1 to 1:4.5, most preferably from 1.5:1 to 1:4.1. Preferably the total amount of the antioxidant(s) is chosen in such a way so that its/their final amount in the formulation is preferably ≤ 7.5 weight-%, more preferably its/their final amount is ranging from 0.5 to 7.5 weight-%, based on the total weight of a) to f) in the formulation. If DL-alpha-tocopherol is used as sole antioxidant, it is preferably used in an amount of at least 1 weight-%, more preferably in an amount ranging from 1.0 to 5.0 weight-%, based on the sum of the weight of the ingredients a) to f). It is assumed that the amount of other tocopherols or analogues thereof, as well as of tocotrienols and analogues thereof is in a similar range as for DL-alpha- tocopherol. Anti-caking agent e) Suitable organic anti-caking agents are talc, cellulose, microcrystalline cellulose, cellulose derivatives or fibres, ferric ammonium citrate, sodium salts of fatty acids such as e.g. sodium stearate, potassium salts of fatty acids such as e.g. potassium stearate, calcium salts of fatty acids such as e.g. calcium stearate, magnesium salts of fatty acids such as e.g. magnesium stearate, aluminum salts of fatty acids such as e.g. aluminum stearate, ammonium salts of fatty acids such as e.g. ammonium stearate, and any mixture of any of them. Other suitable anti-caking agents are inorganic anti-caking agents such as e.g. silicic acid H 2n + 2 Si n O 3n+1 and alkaline/earth alkali metal salts thereof, precipitated silicic acid, silica (= silicon dioxide), modified silica, hydrophobically modified silica, precipitated silica, magnesium oxide, dicalcium diphosphate, tricalcium phosphate, magnesium phosphate, sodium carbonate, potassium carbonate, calcium carbonate, magnesium carbonate, calcium oxide, magnesium oxide, potassium silicate, calcium silicate, magnesium silicate, magnesium trisilicate, aluminum silicate, sodium aluminum silicate, potassium aluminum silicate, calcium aluminum silicate, zeolithe (aluminosilicates), disodium sulfate or mixtures thereof. Also, mixtures of organic and inorganic anti-caking agents may be used. The preferred anti-caking agents used in the formulations of the present invention are anti-caking agents with a particle size D(v,50%) from 100 nm to 10 μm, preferably from 100 nm to 9 μm, more preferably from 150 nm to 5 μm, measured as dry dispersion with a Malvern MasterSizer 3000 (laser diffraction). The particle size of the anti-caking agent can be determined with a laser diffraction system e.g. Malvern MasterSizer 3000, either as dry dispersion or as wet dispersion in oil or Volasil (a mixture of volatile and cyclic silicones such as octamethylcyclo- tetrasiloxane and decamethylcyclopentasiloxane). The particle size can also be determined with electron microscopy. The more preferred anti-caking agents are silicic acid H 2n + 2 Si n O 3n+1 , silica, microcrystalline cellulose, as well as any mixture thereof. The most preferred anti-caking agents are hydrophilic precipitated silicic acid H 2n + 2 Si n O 3n+1 , hydrophilic precipitated silica and any mixture thereof. Thus, the term “anti-caking agent” does preferably not comprise titanium dioxide. Thus, the animal-free formulation of the present invention as well as the milk replacer according to the present invention does not comprise titanium dioxide. The amount of the anti-caking agent e) is preferably ranging from 0.01 to 10 weight-%, based on the sum of the weight of the ingredients a) to f). The more preferred amount of the anti-caking agent is dependent on the drying technique and disclosed below. Characteristics of the formulations of the present invention The formulations of the present invention show preferably a bulk density ranging from 0.4 to 0.9 g/cm 3 , more preferably a bulk density ranging from 0.6 to 0.7 g/cm 3 . Furthermore, the formulations of the present invention show preferably a tap density ranging from 0.7 to 0.85 g/cm 3 . The bulk density and the tap density are measured as follows: A 250 ml glass cylinder is filled with the sample. The volume and the weight are measured. Bulk density is the weight divided by the volume. For measuring the tap density, the sample is tapped with a 2000 Taps with a Stampfvolumeter JEL STAV II (J. Engelsmann AG). The tapped volume is measured. Tap density is the weight divided by the tapped volume. Particle size and measurement To determine the particle size of the animal-free formulation of the present invention a sieve test can be performed with filters of 850 μm pores (mesh 20), 425 μm pores (mesh 40) and 150 μm pores (mesh 100). Furthermore, the particle size of the animal-free formulation may also be determined by laser diffraction analytic whereby the dry dispersion of the sample is measured with a Malvern Mastersizer 2000 or 3000 and Fraunhofer calculation. The particle sizes of the formulation, when determined with laser diffraction with a Malvern Mastersizer 3000 and Fraunhofer calculation, are as follows: D(v,10%) = 1-60 μm; D(v, 50%) = 80-140 μm; D(v, 90%) = 180-240 μm. Synonyms of “D(v,10%)” are “d(v,10%)”, “d(v,10)”, “d(v,0.1)”, “D(v,10)” and “D(v,0.1)”. This applies in an analogous manner for “D(v,50%)” and “D(v,90%)”. Preferred embodiments of the formulations of the present invention Table I and Table II show preferred formulations of the present invention and their composition. Instead of Vitamin A acetate another Vitamin A C1-20 alkyl ester, preferably another Vitamin A C 2-16 alkyl ester, may also be used. The formulations preferably only contain the ingredients cited in the corresponding tables and no further ingredients. Thus, the formulations preferably consist only of the ingredients as cited in the corresponding tables. All amounts in Table I and II except residual moisture are given in weight-%, sum up to 100 weight-% and are based on the total weight of all ingredients except residual moisture. The amount of moisture in Table I and II is given in weight-% and based on the total weight of the formulation. The formulations according to Table I comprise vitamin A acetate and optionally vitamin D3, Gum Arabic; a starch hydrolysate with a DE ≤ 40, preferably with a DE ≤ 30, more preferably with a DE ≤ 25; one or more antioxidants, an anti-caking agent and optionally residual moisture. When vitamin D 3 is present, an oil in the amounts as given above is preferably also present. Preferably the weight ratio of Gum Arabic to the starch hydrolysate with a DE ≤ 40 is ranging from 1:1.3 to 1:3, more preferably it is ranging from 1:1.5 to 1:2.6. The formulations according to Table II comprise vitamin A acetate and optionally vitamin D3, Gum Arabic, modified food starch, a disaccharide, one or more antioxidants, an anti-caking agent and optionally residual moisture. When vitamin D 3 is present, an oil in the amounts as given above is preferably also present. Preferably the weight ratio of modified food starch to the disaccharide is ranging from 1:2 to 2:1, more preferably from 1:1.5 to 1.5:1, most preferably from 1:1.3 to 1.3:1. Table I: Table II
Process for the manufacture of the formulation The formulation of the present invention is preferably manufactured according to a process comprising the following steps: A) Dissolving Gum Arabic b), the ingredient(s) c) and, if present, a water- soluble antioxidant(s) d) in water to obtain a matrix; B) Heating the fat-soluble vitamin(s) a), the fat-soluble antioxidant(s) d) and, if present, the oil f), to obtain an active phase; C) Emulsifying the active phase obtained in step B) into the matrix obtained in step A) to obtain a dispersion; D) Drying the dispersion obtained in step C), optionally in presence of an anti-caking agent, to obtain the formulation. The single steps are disclosed in more detail below. Further details, that may also be generalized, are given in the examples. Step A) The amounts of the Gum Arabic b), the ingredient(s) c) and if present, the water- soluble antioxidant(s) c) are chosen so that the final amounts of these compounds in the animal-free formulation after having performed steps A) to D) is as described above. Step A) is preferably performed at a temperature ranging from 45 to 80°C, more preferably ranging from 50 to 78°C, most preferably ranging from 55 to 75°C. Step B) The amounts of the fat-soluble vitamin(s) a), the fat-soluble antioxidant(s) d) and, if present, the oil f) are chosen so that the final amounts of these compounds in the animal-free formulation after having performed steps A) to D) is as described above. Step B) is preferably performed at a temperature to bring the components a) and d) into a liquid state. When vitamin D, preferably vitamin D 3 , is present in the formulation of the present invention, the vitamin D is preferably added to the other fat-soluble vitamin and the fat-soluble antioxidant as oily suspension, whereby the weight ratio of vitamin D to the oil is preferably ranging from 1:1 to 1:10, more preferably from 1:2 to 1:5. Step C) Preferably this step is performed at a mixing temperature in the range of from 40 to 78°C, more preferably at a mixing temperature in the range of 50 to 75°C, even more preferably at a mixing temperature in the range of 55°C to 70°C to obtain a dispersion. The homogenization can be achieved by using a rotor-stator device or a high- pressure homogenizer or both. Other devices known to the person skilled in the art may also be used. Step D) The dividing and drying of the mixture of the oil-in-water preparation to produce the animal-free formulation according to the present invention can be done in any conventional way, such as spray cooling, modified spray cooling, spray drying, spray-drying in combination with fluidized bed granulation, modified spray drying or sheet drying and crushing, see e.g. WO 91/06292 A1. Furthermore, the conversion to the animal-free formulation can be achieved by a powder-catch technique, whereby the sprayed dispersion droplets are caught by the anti-caking agent (so-called “catch media”), and dried. If spray drying is carried out as drying step, preferably an anti-caking agent is added in an amount so that the amount in the final formulation is at most 1.5 weight-%, preferably the amount is ranging from 0.01 to 1.0 weight-%, based on the sum of the weight of the ingredients a) to f) together. If a powder catch process is carried out as drying step, preferably an anti-caking agent is added in an amount so that the amount in the final formulation is at least 5.0 weight-%, preferably the amount is ranging from 5.0 to 10.0 weight-%, more preferably the amount is ranging from 5.0 to 7.0 weight-%, based on the sum of the weight of the ingredients a) to f) together. Further embodiments of the present invention Use The present invention is also directed to the use of the formulation according to the present invention with the preferences as given above as additive to milk replacers, especially for young non-human mammals such as calves, foals, young goats, young camels and young sheep. Milk replacer according to the present invention The present invention is also directed to a milk replacer comprising the formulation according to the present invention with the preferences as given above. “Milk replacer” means any substance or product, including additives, whether processed, partially processed or unprocessed, intended to be used to replace the milk of non-human mammals such as calves, foals, young sheep, camel foals and young goats. Such milk replacer preferably further contains essential nutrients including proteins, amino acids, carbohydrates, fats, vitamins and minerals The amount of the animal-free formulation and the fat-soluble vitamin respectively in such milk replacers follows the regulatory guidelines in the regions depending on the specific animal species and its age. In the Supplementation Guidelines the amount of the vitamins A and D 3 is given in International Units (“I.U.”). To ensure that the active content in the feed is communicated in a systematic way, “I.U.” is used as a universal unit for fat soluble vitamins because there are different forms of the vitamins with varying amounts of fat-soluble vitamins. The formulation according to the present invention is usually added to milk replacers in form of a premix, i.e. a mixture with other micro-nutrients such as other vitamins or their formulations and minerals. The premix inclusion in feed is < 1 weight-% for many species. It can, however, also be added as such. The amount of the formulation according to the present invention needed to be included into the milk replacer is calculated based on the active content of the milk replacer and the targeted dosage of the fat-soluble vitamin considering said inclusion level. The conversion factors of the fat-soluble vitamins are as follows: 1 I.U. Vitamin A corresponds to 0.344 μg of Vitamin A acetate; 1 I.U. Vitamin D 3 corresponds to 0.025 μg of Vitamin D 3 ; 1 g of Vitamin K 3 (menadione) corresponds to 2.0 g of menadione sodium bisulfite (MSB) or to 2.3 g of menadione nicotin-amide bisulfite (MNB). The following Table III shows the amounts of the added fat-soluble vitamins. The exact amount is depending on several factors such as phase/age of the animal, animal species and legal local limits. Table III 1) Amount added per kg of calf milk replacer powder. 2) Supplementary amount per animal per day. The invention is now further illustrated in the following non-limiting examples. Examples The following examples 1 and 2 illustrate the manufacture of the formulation of the present invention. Examples 1 and 2 (Table 1) The matrix components, i.e. Gum Arabic, the ingredient(s) c) and the water-soluble antioxidant are dissolved in water at approximately 70°C to obtain the “matrix”. Vitamin A acetate, the fat-soluble antioxidant and, if present, the oil are heated at approximately 65°C under stirring until complete melting of vitamin A acetate (“active phase”). They are then emulsified into the matrix. Hereby, the amounts of the ingredients are chosen in such a way that their concentrations in the final formulation are as disclosed in Table 1 (n.d. = not determined). After thorough mixing the resulting dispersion is sprayed into a spray tower optional in the presence of an anti-caking agent to form droplets of the desired size. The solidified droplets are then dried by a drying air of various temperatures (5-75°C). Example 1: If spray-drying is carried out: The dried powder is sieved through a 1 mm sieve to discard agglomerates. Example 2: If powder-catch is carried out: The dried powder is separated from the majority of the anti-caking agent and sieved through 150 μm and 600 μm filters. The particle size of the dried powder is determined with laser diffraction analytic with a Malvern Mastersizer 3000. The sample is hereby dry dispersed. By applying the Fraunhofer theory the particle size distribution of the sample is calculated. Furthermore, the bulk and the tap density are measured according to the procedures as described above. Table 1