Genipin-rich material and its use

Abstract

A method of preparing genipin-rich materials from the fruit of Genipa americana fruit for their use as a cross-linking agent and as a raw material to produce colors is disclosed. The genipin-rich materials can be used in a broad range of applications including personal care, cosmetics, dietary supplements, packaging, textiles, beverages, foodstuffs, drugs, and animal feeds.

What is claimed is: 
     
       1. A method of preparing a genipin-rich red colorant from Genipa americana comprising:
 (a) extracting a Genipa americana fruit source selected from the group consisting of Genipa americana whole fruit, fruit juice, fruit puree, fruit juice concentrate, dried powder forms of the fruit and juice, water-insoluble parts of the fruit, and mixtures thereof, with a solvent selected from water, organic solvents, and mixtures thereof, to produce a genipin-rich extract; 
 (b) hydrolyzing the genipin-rich extract for 30 minutes to 24 hours to produce a hydrolyzed extract; 
 (c) adjusting the pH of the hydrolyzed extract to about 3.5-5.0, to produce an acidified solution; 
 (d) heating the acidified solution to 65 to 85° C. for about 5 to 120 minutes to produce a brown-colored precipitate and removing the precipitate to produce a clear solution, 
 (e) adjusting the pH of the clear solution to about 4-4.6, 
 (f) mixing the solution of step (e) with one or more compounds having a primary amine group in the presence of an organic acid or salt thereof, under anaerobic conditions, for a sufficient time and temperature to produce the genipin-rich red colorant. 
 
     
     
       2. The method according to  claim 1  wherein the colorant is further purified and concentrated using an ion exchange column and/or a membrane filter. 
     
     
       3. The method according to  claim 1  wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, glutamic acid, and mixtures thereof. 
     
     
       4. The method of  claim 1  wherein the organic solvent is selected from ethyl acetate, butyl acetate, n-butanol, diethyl ether, hexane, 2-butanone, chloroform, 1,2-dichloroethane, xylene, methyl-t-butyl ether, toluene, carbon tetrachloride, trichloroethylene, cyclohexane, pentane, heptane and mixtures thereof. 
     
     
       5. The method according to claim 1 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, aspartic acid, glutamic acid, glycine, histidine, valine, leucine, serine, and mixtures thereof. 
     
     
       6. A method of preparing a genipin-rich colorant from Genipa americana comprising:
 (a) blending water and a Genipa americana fruit source selected from the group consisting of Genipa americana whole fruit, fruit pulp, fruit juice, fruit puree, fruit juice concentrate, dried powder forms of the fruit or juice, water-insoluble parts of the fruit, and leaves from Genipa Americana L., and mixtures of any thereof to produce a water and Genipa americana fruit source mixture;   (b) mixing the water and Genipa americana fruit source mixture to produce a puree;   (c) removing at least a portion of insoluble solid material from the puree to obtain a filtrate of the insoluble solid material;   (d) adjusting the pH of filtrate to a pH of 3.5-5.0;   (e) concentrating the filtrate;   (f) extracting the concentrated filtrate using an organic solvent to obtain a solvent extract layer, wherein the organic solvent is either polar or non-polar, wherein when the organic solvent is polar, it is selected from the group consisting of acetic acid, ethanol, n-propanol, iso-propanol, methanol, or acetic, citric, and phosphoric acid buffer solutions, and mixtures thereof, and when the organic solvent is non-polar, it is selected from the group consisting of ethyl acetate, butyl acetate, and n-butanol, and mixtures thereof;   (g) removing at least some of the organic solvent from the layer to obtain a genipin-rich extract comprising at least 70% w/w of genipin; and   (h) mixing the genipin-rich extract of step (g) with one or more compounds having a primary amine group to produce the genipin-rich colorant, the genipin-rich colorant having a different color than the genipin-rich extract.   
     
     
       7. The method according to claim 6 wherein the Genipa americana fruit source comprises peeled and cut fruit. 
     
     
       8. The method according to claim 6 wherein in step (a), the water is deionized water. 
     
     
       9. The method of claim 6 wherein in step (a), the blending comprises blending the water and Genipa americana fruit source in ratio in the range of about 1.0:0.67 to about 1.0:0.86 by weight. 
     
     
       10. The method of claim 6 wherein in step (b), the puree is allowed to stand for at least about 15 minutes. 
     
     
       11. The method of claim 6 wherein step (c) comprises a first filtering step to form a first filtrate, followed by adding water to the first filtrate, mixing the water and first filtrate, and then a second filtering step to form a second filtrate, and step (d) comprises adjusting the pH of the second filtrate. 
     
     
       12. The method of claim 11 wherein the water added to the first filtrate is deionized water. 
     
     
       13. The method of claim 11 wherein the mixing of the water and the first filtrate is performed for a period of at least about 15 minutes. 
     
     
       14. The method of claim 6 wherein the acid is selected from the group consisting of acetic acid, formic acid, lactic acid, citric acid, tartaric acid, adipic acid, oxalic acid, succinic acid, fumaric acid, malic acid, and phosphoric acid, and mixtures thereof. 
     
     
       15. The method of claim 6 wherein the acid is selected from citric acid and tartaric acid, and mixtures thereof. 
     
     
       16. The method of claim 6 wherein step (e) comprises concentrating the filtrate using rotary evaporation. 
     
     
       17. The method of claim 6 wherein step (f) comprises extracting the concentrated filtrate using the organic solvent in a ratio of about 1:1 by weight. 
     
     
       18. The method of claim 6 wherein the removing in step (g) comprises evaporating at least a portion of the organic solvent from the layer. 
     
     
       19. The method of claim 18 wherein the evaporating is performed using rotary evaporation. 
     
     
       20. The method of claim 6 wherein step (f) comprises repeating the liquid-liquid extraction to obtain more than one solvent extract layer. 
     
     
       21. The method of claim 20 further comprising combining the solvent extract layers, and wherein in step (g), the removing comprises evaporating at least a portion of the organic solvent from the combined solvent extract layers. 
     
     
       22. The method of claim 21 wherein the evaporating is performed at a temperature of about 40° C. to about 55° C. and a vacuum of about 35 mm Hg. 
     
     
       23. The method of claim 22 wherein the evaporating is performed using rotary evaporation. 
     
     
       24. The method of claim 6 wherein at least a portion of organic solvent removed in step (g) is reused for extracting in step (f). 
     
     
       25. The method of claim 6 wherein the evaporating is performed at a temperature of about 40° C. to about 55° C. 
     
     
       26. The method of claim 25 wherein the evaporating is performed at a vacuum of about 35 mm Hg. 
     
     
       27. The method of claim 6 wherein the organic solvent is polar. 
     
     
       28. The method of claim 6 wherein the organic solvent is non-polar. 
     
     
       29. The method of claim 28 wherein the non-polar organic solvent is an acetate. 
     
     
       30. The method of claim 29 wherein the acetate is ethyl acetate. 
     
     
       31. The method of claim 30 wherein the evaporating is performed using rotary evaporation, at a temperature of about 40° C., and at a vacuum of about 35 mm Hg, and wherein the genipin-rich extract obtained in step (g) comprises at least about 74.4% w/w of genipin. 
     
     
       32. The method of claim 29 wherein the acetate is butyl acetate. 
     
     
       33. The method of claim 32 wherein the evaporating is performed using rotary evaporation, at a temperature of about 55° C., and at a vacuum of about 35 mm Hg, and wherein the genipin-rich extract obtained in step (g) comprises at least about 90.9% w/w of genipin. 
     
     
       34. The method of claim 29 wherein the acetate removed in step (g) is reused for extracting in step (f). 
     
     
       35. The method according to claim 6,
 wherein after the removing step (g) and before the mixing step (h):
 hydrolyzing the genipin-rich extract for about 30 minutes to 24 hours to produce a hydrolyzed extract; 
 adjusting the pH of the hydrolyzed extract to about 3.5-5.0, to produce an acidified solution; 
 heating the acidified solution to about 65 to 85° C. for about 5 to 120 minutes to produce a brown-colored precipitate and removing the precipitate to produce a clear solution comprising the genipin-rich extract; and 
 adjusting the pH of the clear solution comprising the genipin-rich extract to about 4.0-4.6; 
   wherein the mixing in step (h) comprises mixing the clear solution comprising the genipin-rich extract with the one or more compounds having a primary amine group, and the mixing is performed in the presence of an organic acid or salt thereof, under anaerobic conditions, for a sufficient time and temperature to produce the genipin-rich colorant, wherein the genipin-rich colorant is red.   
     
     
       36. The method according to claim 35 wherein the colorant is further purified and concentrated using an ion exchange column and/or a filter. 
     
     
       37. The method according to claim 35 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, glutamic acid, and mixtures thereof. 
     
     
       38. The method according to claim 35 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, aspartic acid, glutamic acid, glycine, histidine, valine, leucine, serine, and mixtures thereof. 
     
     
       39. The method according to claim 6,
 wherein after the removing step (g) and before the mixing step (h):
 dissolving the genepin-rich extract in a solution comprising ethanol to form a genepin-rich extract and ethanol solution; 
 mixing a hydroxide with the genepin-rich extract and ethanol solution to form a hydroxide-extract-ethanol mixture; 
 hydrolyzing the hydroxide-extract-ethanol mixture to form a hydrolyzed extract; 
 adjusting the pH of the hydrolyzed extract to 3.5-5.0 to produce an acidified solution; 
 heating the acidified solution to about 65 to 85° C. for about 5 to 120 minutes; 
 allowing the heated acidified solution to cool; 
 removing precipitate from the cooled acidified solution; 
 adjusting the pH of the cooled acidified solution to about 5.0-7.5 and removing formed precipitate therefrom to produce a solution comprising the genipin-rich extract; and 
 adjusting the pH of the solution comprising the genipin-rich extract to about 4.0-4.6; 
   wherein the mixing in step (h) comprises mixing the solution comprising the genipin-rich extract with one or more compounds having a primary amine group, and the mixing is performed in the presence of an organic acid or salt thereof, for a sufficient time and temperature, to produce the genipin-rich colorant, wherein the genipin-rich colorant is red.   
     
     
       40. The method according to claim 39 wherein the colorant is further purified and concentrated using an ion exchange column and/or a filter. 
     
     
       41. The method according to claim 39 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, glutamic acid, and mixtures thereof. 
     
     
       42. The method according to claim 39 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, aspartic acid, glutamic acid, glycine, histidine, valine, leucine, serine, and mixtures thereof. 
     
     
       43. The method according to claim 6,
 wherein after the removing step (g) and before the mixing step (h):
 dispersing the genipin-rich extract in an ethanol aqueous solution to produce a genipin-rich extract in ethanol aqueous solution; 
 heating the genipin-rich extract in ethanol aqueous solution for a sufficient time and temperature to dissolve solids to produce a genipin-rich extract in ethanol aqueous solution with dissolved solids; 
   wherein the mixing in step (h) comprises mixing the genipin-rich extract in ethanol aqueous solution with dissolved solids with the one or more compounds having a primary amine group for a sufficient time and temperature to produce the genipin-rich colorant, wherein the genipin-rich colorant is green.   
     
     
       44. The method according to claim 43 wherein the colorant is further purified and concentrated using an ion exchange column and/or a filter. 
     
     
       45. The method according to claim 43 wherein the primary amine group is an amino acid selected from isoleucine, threonine, cysteine, tryptophan, and mixtures thereof. 
     
     
       46. The method according to claim 43 wherein primary amine group is an amino acid selected from isoleucine, threonine, or mixtures thereof. 
     
     
       47. The method according to claim 6,
 wherein after the removing step (g) and before the mixing step (h):
 dispersing the genipin-rich extract in an ethanol aqueous solution to produce a genipin-rich extract in ethanol aqueous solution; 
 heating the genipin-rich extract in ethanol aqueous solution for a sufficient time and temperature to dissolve solids to produce a genipin-rich extract in ethanol aqueous solution with dissolved solids; 
   wherein the mixing in step (h) comprises mixing the genipin-rich extract in ethanol aqueous solution with the dissolved solids with a compound having a primary amine group, wherein the primary amine group is an amino acid, wherein the mixing is performed in the presence of oxygen, wherein the molar ratio of genipin-rich extract to the amino acid is from about 1:0.5 to about 1:10, for a sufficient time and temperature to produce to produce the genipin-rich colorant, wherein the genipin-rich colorant is blue.   
     
     
       48. The method according to claim 47, wherein after the mixing in step (h), heating to accelerate reaction speed between the genipin in the genipin-rich extract and the amino group. 
     
     
       49. The method according to claim 47 wherein the amino acid selected from lysine, histidine, arginine, glutamine, asparagine, methionine, glycine, glutamic acids, tyrosine, valine, alanine, serine, leucine, taurine, carnitine, ornithine, citrulline, and mixtures thereof. 
     
     
       50. The method of claim 47 wherein the amino acid is histidine. 
     
     
       51. The method according to claim 6,
 wherein after the removing step (g) and before the mixing step (h):
 dispersing the genipin-rich extract in an ethanol aqueous solution to produce a genipin-rich extract and ethanol aqueous solution; 
   wherein the mixing step (h) comprises mixing the genipin-rich extract and ethanol aqueous solution with a pentose and a compound having a primary amine group, wherein the compound having a primary amine group is an amino acid selected from alanine, arginine, lysine, aspartic acid, glutamic acid, glycine, histidine, valine, leucine, serine, and mixtures thereof, for a sufficient time and temperature to produce the genipin-rich colorant, wherein the genipin-rich colorant is orange-red.   
     
     
       52. The method according to claim 51 wherein the colorant is further purified and concentrated using an ion exchange column and/or a filter. 
     
     
       53. The method according to claim 51 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, glutamic acid, and mixtures thereof. 
     
     
       54. The method according to claim 51 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, aspartic acid, glutamic acid, glycine, histidine, valine, leucine, serine, and mixtures thereof. 
     
     
       55. The method of claim 6,
 wherein after the removing step (g) and before the mixing step (h):
 dissolving the genepin-rich extract in a solution comprising ethanol to form a genepin-rich extract and ethanol solution; 
 mixing a hydroxide with the genepin-rich extract and ethanol solution to form a hydroxide-extract-ethanol mixture; 
 hydrolyzing the hydroxide-extract-ethanol mixture to form a hydrolyzed extract; 
 adjusting the pH of the hydrolyzed extract to 3.5-5.0 to produce an acidified solution; 
 heating the acidified solution to about 65 to 85° C. for about 5 to 120 minutes; 
 allowing the heated acidified solution to cool; 
 removing precipitate from the cooled acidified solution; 
 adjusting the pH of the cooled acidified solution to about 6.0-7.0 and removing formed precipitate therefrom to produce a solution comprising the genipin-rich extract; and 
 adjusting the pH of the solution comprising the genipin-rich extract to about 4.0-4.6; 
   wherein the mixing step (h) comprises mixing the solution comprising the genipin-rich extract with one or more compounds having a primary amine group, taurine, and an alkaline earth metal, or metals with multi-valences for a sufficient time and temperature to produce the genipin-rich colorant, wherein the genipin-rich colorant is dark brown-black.   
     
     
       56. The method according to claim 55 wherein the colorant is further purified and concentrated using an ion exchange column and/or a filter. 
     
     
       57. The method according to claim 55 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, glutamic acid, and mixtures thereof. 
     
     
       58. The method according to claim 55 wherein the one or more compounds having a primary amine group is an amino acid selected from alanine, arginine, lysine, aspartic acid, glutamic acid, glycine, histidine, valine, leucine, serine, and mixtures thereof.