Method of Preparing a Glycoside of a Mono or Diacylglycerol Product From a Plant Material

Abstract

The present invention provides a method of preparing a glycoside of a mono- or diacylglycerol product from a plant material. The method comprises the steps of: (i) optionally milling the plant material, (ii) extracting the optionally milled plant material with a first aqueous extraction solution obtaining a first liquid phase and a first solid phase, (iii) separating the liquid phase from the solid phase to obtain a glycoside of mono- or diacylglycerol product, (iv) extracting the solid phase from step (iii) with a second extraction solution obtaining a second liquid phase and a second phase, and (v) separating the second liquid phase from the second solid phase to obtain a second and a third glycoside of mono- or diacylglycerol product, wherein the second extraction solution further comprises a cell wall degrading enzyme or a mixture of cell wall degrading enzymes.

1 . A method of preparing a glycoside of a mono- or diacylglycerol product from a plant material, said method comprises the steps of:
 (i) optionally milling the plant material,   (ii) extracting the optionally milled plant material with a first aqueous extraction solution obtaining a first liquid phase and a first solid phase,   (iii) separating the liquid phase from the solid phase to obtain a glycoside of mono- or diacylglycerol product.   
   
   
       2 . A method according to  claim 1 , wherein the liquid phase comprises a first glycoside of mono- or diacylglycerol product and the first glycoside of mono- or diacylglycerol product is in the form of a juice product. 
   
   
       3 . A method according to any one of the preceding claims, wherein the first aqueous extraction solution and the plant material is mixed at a ratio of 100:1 to 1:100, preferably at the ratio of 50:1 to 1:50, such as 25:1 to 1:25, preferably at the ratio of 10:1. 
   
   
       5 . A method according to any one of the preceding claims, wherein the extraction in step (ii) is performed at a first extraction temperature in the range of 20-110° C., such as in the range of 40-100° C., e.g. in the range of 60-95° C., such as in the range of 75-90° C., e.g. 90° C. 
   
   
       6 . A method according to any one of the preceding claims, wherein the extraction in step (ii) is performed in a period ranging from 10 seconds to 20 minutes, such as ranging from 30 seconds to 10 minutes, e.g. ranging from 45 seconds to 5 minutes, such as ranging from 1 minute to 2 minutes, e.g. 1 minute. 
   
   
       7 . A method according to any one of the preceding claims, wherein the method further comprises the step of:
 (iv) extracting the solid phase obtained from step (iii) with a second extraction solution obtaining a second liquid phase and a second solid phase.   
   
   
       8 . A method according to  claim 7 , wherein the second aqueous extraction solution and the plant material is mixed at a ratio of 50:1 to 1:50, preferably at the ratio of 25:1 to 1:25, such as 10:1 to 1:10, preferably at the ratio of 5:1. 
   
   
       9 . A method according to any one of  claims 7 - 8 , wherein the extraction in step (iv) is performed at a second extraction temperature in the range of 20-75° C., such as in the range of 30-60° C., e.g. in the range of 40-55° C., such as in the range of 45-50° C., e.g. 50° C. 
   
   
       10 . A method according to any one of  claims 7 - 9 , wherein the pH of the extraction in step (iv) is adjusted to a pH value in the range of pH 1-10, such as in the range of pH 2-9, e.g. in the range of pH 3-7, such as in the range of pH 4-6, e.g. pH 4.5. 
   
   
       11 . A method according to  claim 10 , wherein the pH is adjusted with NaOH. 
   
   
       12 . A method according to any one of  claims 7 - 11 , wherein the second extraction solution further comprises a cell wall degrading enzyme or a mixture of cell wall degrading enzymes. 
   
   
       13 . A method according to  claim 12 , wherein the cell wall degrading enzyme is one or more enzymes selected from the group consisting of cellulase, β-glucanase, xylanase, arabanase and pentosanase. 
   
   
       14 . A method according to  claims 12  or  13 , wherein the enzymatic activity is inactivated before obtaining (a) further isolated glycoside of mono- or diacylglycerol product(s). 
   
   
       15 . A method according to  claim 14 , wherein the enzymatic activity is inactivated by heat or by addition of an acid or a base. 
   
   
       16 . A method according to any one or  claims 7 - 15 , wherein the method further comprises the step of:
 (v) separating the second liquid phase from the second solid phase to obtain a second and a third glycoside of mono- or diacylglycerol product.   
   
   
       17 . A method according to  claim 16 , wherein the second liquid phase comprises a second glycoside of mono- or diacylglycerol product and is in the form of a tincture product. 
   
   
       18 . A method according to any one of  claims 16  or  17 , wherein the second solid phase comprises a third glycoside of mono- or diacylglycerol product and is in the form of a solid concentrate. 
   
   
       19 . A method according to any one of the preceding claims, wherein the glycoside of mono- or diacylglycerol product is in the form a juice product, a tincture product and/or a solid concentrate. 
   
   
       20 . A method according to any one of  claims 18  or  19 , wherein the solid concentrate is dried. 
   
   
       21 . A method according to any one of the preceding claims, wherein the separation in step (iii) and/or in step (vii) is performed by decanting, centrifugation or filtration. 
   
   
       22 . A method according to any one of the preceding claims, wherein the method does not involve the use of organic solvents. 
   
   
       23 . A method according to any one of the preceding claims, wherein the first and/or second aqueous extraction solution is water. 
   
   
       24 . A method according to any one of the preceding claims, wherein the plant material is rose hip such as  Rosa  canina (“dog rose-hip”),  Rosa galica, Rosa condita, Rosa rugosa, Rosa hugonis, Rosa nitida, Rosa pendulina, Rosa pimpinellifolia , and  Rosa sericea.    
   
   
       25 . A method according to any one of  claims 19 - 24 , wherein the first, second and third glycoside of mono- or diacylglycerol product is produced in one combined process of all steps (i) to step (v). 
   
   
       26 . A method according to any one of  claims 1 - 25 , wherein the glycoside of mono- or diacylglycerol product comprises a glycoside of mono- or diacylglycerol compound having the following formula I: 
     
       
         
         
             
             
         
       
     
     wherein R and R′ independently are selected from hydrogen, C 10-24 -alkyl, and C 10-24 -acyl, said alkyl and acyl groups having 0 to 5 unsaturated bonds, and R 1 , R 2 , R 3  and R 4  independently are selected from hydrogen and glycoside moieties; with the first proviso that not both of R and R′ are hydrogen, and with the second proviso that none of R and R′ is eicosapentaenoyl. 
   
   
       27 . A method according to  claim 26 , wherein any alkyl and acyl groups have 0 to 4 unsaturated bonds, such as 1-3 unsaturated bonds, e.g. 2 or 3 unsaturated bonds, in particular 3 unsaturated bonds. 
   
   
       28 . A method according to any of  claims 26  or  27 , wherein any unsaturated bonds are double bonds. 
   
   
       29 . A method according to any of  claims 26 - 28 , wherein R and R′ both are C 16-20 -acyl having 1 to 3 double bonds, such as C 1-8 -acyl having 3 double bonds. 
   
   
       30 . A method according to any of  claims 26 - 29 , wherein the compound has the formula II: 
     
       
         
         
             
             
         
       
     
     wherein R, R′, R 1 , R 2 , R 3  and R 4  all are as defined in any of  claims 1 - 5 . 
   
   
       31 . A method according to any of  claims 26 - 30 , wherein the compound is selected from β- D -galactopyranosyl derivatives, α- D -galactopyranosyl derivatives, β-D-glucopyranosyl derivatives, and α- D -glucopyranosyl derivatives. 
   
   
       32 . A method according to any one of  claims 26 - 31 , wherein the compound is selected from 3-β- D -galactopyranosyloxy-2-(octadeca-9Z,12Z,15Z-trienoyloxy)propanyl octadeca-9Z,12Z,15Z-trienoate, 3-β- D -glucopyranosyloxy-2-(octadeca-9Z,12Z,15Z-trienoyloxy)propanyl octadeca-9Z,12Z,15Z-trienoate, 3-α- D -galactopyranosyloxy-2-(octadeca-9Z,12Z,15Z-trienoyloxy)propanyl octadeca-9Z,12Z,15Z-trienoate, and 3-α- D -glucopyranosyloxy-2-(octadeca-9Z,12Z,15Z-trienoyloxy)propanyl octadeca-9Z,12Z,15Z-trienoate. 
   
   
       33 . A juice product comprising the glycoside of mono- or diacylglycerol product obtainable by the method according to  claims 1 - 32 . 
   
   
       34 . A tincture product comprising the glycoside of mono- or diacylglycerol product obtainable by the method according to  claims 1 - 32 . 
   
   
       35 . A solid concentrate comprising the glycoside of mono- or diacylglycerol product obtainable by the method according to  claims 1 - 32 . 
   
   
       36 . A product comprising the juice product according to  claim 33 , the tincture product according to  claim 34  or the solid concentrate according to  claim 35 . 
   
   
       37 . A product according to  claim 36 , wherein the product is a food product, a natural medicine product or a dietary supplement. 
   
   
       38 . A use of the juice product according to  claim 33 , the tincture product according to  claim 34  or the solid concentrate according to  claim 35  for the preparation of a medicament for the treatment, alleviation or prophylaxis of inflammatory conditions in a mammal. 
   
   
       39 . A use according to  claim 38 , wherein the medicament is adapted for oral, buccal, parenteral, topical, rectal, transdermal or intranasal administration. 
   
   
       40 . A use according to any of  claims 38  or  39 , wherein the medicament is in the form of a daily dose form applicable for administration of about 0.005-50 mg/kg body weight per day, in unitary or multiple doses. 
   
   
       41 . A use according to any of  claims 38 - 40 , wherein the inflammatory condition is selected from hepatitis, meningitis, rheumatoid arthritis, inflammatory bowl diseases, allergic syndromes, diabetes, congestive heart disease, psoriatic, reactive or osteo-arthritis or other arthritides, multiple sclerosis, sepsis/septic shock, dermal inflammation, graft rejection, and inflammation secondary to chemotherapy or radiotherapy of neoplastic disease. 
   
   
       42 . A use according to any of  claims 38 - 41 , wherein the inflammatory condition is arthritis. 
   
   
       43 . A use according to any of  claims 38 - 42 , wherein the inflammatory condition is osteoarthrosis. 
   
   
       44 . A method of treating, alleviating, or preventing inflammatory conditions in a mammal in need therefore, said method comprising the step of administering a medicament or a product comprising the juice product according to  claim 33 , the tincture product according to  claim 34  or the solid concentrate according to  claim 35 . 
   
   
       45 . A method according to  claim 44 , wherein the medicament is adapted for oral, buccal, parenteral, topical, rectal, transdermal or intranasal administration. 
   
   
       46 . A method according to any of  claims 44  or  45 , wherein the medicament is in the form of a daily dose form applicable for administration of about 0.005-50 mg/kg body weight per day, in unitary or multiple doses. 
   
   
       47 . A method according to any of  claims 44 - 46 , wherein the inflammatory condition is selected from hepatitis, meningitis, rheumatoid arthritis, inflammatory bowl diseases, allergic syndromes, diabetes, congestive heart disease, psoriatic, reactive or osteo-arthritis or other arthritides, multiple sclerosis, sepsis/septic shock, dermal inflammation, graft rejection, and inflammation secondary to chemotherapy or radiotherapy of neoplastic disease. 
   
   
       48 . A method according to any of  claims 44 - 47 , wherein the inflammatory condition is arthritis. 
   
   
       49 . A use according to any of  claims 44 - 48 , wherein the inflammatory condition is osteoarthrosis.