Process for isolating plant extract fractions

• US 5,301,694 A
• Filed: 11/12/1991
• Issued: 04/12/1994
• Est. Priority Date: 11/12/1991
• Status: Expired due to Fees

• Current Assignee: Philip Morris USA Incorporated (Altria Group, Inc.)
• Original Assignee: Philip Morris USA Incorporated (Altria Group, Inc.)
• Inventors: Hale, Robert W., Raymond, Wynn R.
• Primary Examiner(s): PREBILIC, PAUL B
• Application Number:
  US07/789,979
• Time in Patent Office: 882 Days
• Field of Search: 131/297, 131/298, 131/300, 131/301, 131/302, 131/194, 131/199, 131/905, 426/490, 426/495, 426/425, 426/429, 426/430, 426/431
• US Class Current: 131/297
• CPC Class Codes: A24B 15/24 by extraction; Tobacco extracts
• CPC Class Codes: Y10S 131/905 Radiation source for sensing condition or characteristic

Abstract

Fractionated plant extracts, particularly essentially nicotine-free tobacco extracts, useful as tobacco flavorants and methods for preparing and using the same are described herein. These fractionated extracts are stable, non-volatile and relatively odorless under conditions of smoking article or smoking substitute article manufacture and storage, but when thermally provoked, the extracts deliver characteristic flavor to smoking articles or smoking substitute articles. The fractionated plant extracts may be prepared by contacting plant matter with a solvent to produce a crude plant extract; removing the solvent from this plant extract; and isolating a fraction of this extract that when thermally provoked provides the characteristic aroma and flavor of the plant by size exclusion chromatography and monitoring the fraction'"'"'s integrity by suitable detection means.

Claims

1. A process for producing and isolating intermediate molecular weight fractions from plant extracts, which fractions are substantially non-volatile and stable under ambient temperature conditions and are useful as flavoring agents, comprising:

(a) contacting plant matter with a solvent to produce a crude plant extract;

(b) concentrating said crude plant extract;

(c) subjecting said concentrated crude plant extract to a size exclusion process to provide intermediate molecular weight fractions having a molecular weight in the range from about 600 Daltons to about 100,000 Daltons, wherein said size exclusion process separates on the basis of a physical parameter to provide the intermediate molecular weight fractions; and

(d) isolating said intermediate molecular weight fractions by using a detection method to monitor changes in the distribution and intensity of the substances eluted in said size exclusion process.

2. The process according to claim 1, wherein said detection method employs a detection means selected from the group consisting of ultraviolet absorption, infrared absorption, refractive index, light scattering, circular dichroism, pH, and electrochemical conductance.

3. The process according to claim 1, further comprising the step of (e) concentrating said intermediate molecular weight fractions.

4. The process according to claim 1 wherein said plant matter is tobacco.

5. The process according to claim 4 further comprising the step of (e) contacting said intermediate molecular weight fractions from step (d) which contain nicotine detectable by suitable detection means with an ion-exchange material to essentially remove the nicotine.

9. The process according to claim 1, wherein said plant matter comprises matter selected from the group consisting of leaves, stalks, stems, seeds, roots and petals.

10. The process according to claim 1, wherein said plant matter is selected from the group consisting of tobacco, cocoa, coffee, sage, fennel, licorice, cinnamon, foenugreek and any mixture thereof.

11. The process according to claim 4, wherein said tobacco is selected from the group consisting of Flue-Cured, Maryland, Burley, Turkish, and any mixture thereof.

12. The process according to claim 11, wherein said tobacco is cut into sizes of about 10 to about 60 cuts per inch.

13. The process according to claim 12, wherein prior to cutting said tobacco, said tobacco is moistened to a level from about 10% to about 30%.

14. The process according to claim 13, wherein said tobacco is moistened to a level from about 12-13%. )

16. The process according to claim 1, wherein said plant matter is mixed with an inert filler.

17. The process according to claim 16, wherein said inert filler is a member selected from the group consisting of excelsior, glass beads, oat hulls, and any combination thereof.

18. The process according to claim 17, wherein said inert filler is oat hulls.

19. The process according to claim 16, wherein said plant matter and inert filler are present in the ratio of about 90% plant matter to about 10% inert filler.

20. The process according to claim 1, wherein said solvent is selected from the group consisting of water, ethanol, and mixtures thereof.

21. The process according to claim 20, wherein said solvent is a mixture of water and ethanol in the range of from about 95:

5 to about 20;

80 water to ethanol by volume.

22. The process according to claim 21, wherein said solvent is a mixture of water and ethanol in the ratio of from about 1 to 1 by volume to about 1 to 3 by volume.

23. The process according to claim 20, wherein said solvent is water.

24. The process according to claim 23, wherein the water is used to produce said crude plant extract at a temperature of about 20°

C. to about 90°

C.

25. The process according to claim 24, wherein the water is used to produce said crude plant extract at a temperature of about 75°

C. to about 90°

C.

26. The process according to claim 21, wherein the water-ethanol mixture is used to produce said crude plant extract at a temperature of about 20°

C. to 65°

C.

27. The process according to claim 26 wherein the water-ethanol mixture is used to produce said crude plant extract at a temperature of about 25°

C. to about 40°

C.

28. The process according to claim 1, wherein said crude plant extract of step (a) is concentrated in step (b) by a method selected from the group consisting of distillation, reverse osmosis, microfiltration, ultrafiltration, nanofiltration, hollow fiber diafiltration, and any combination thereof.

29. The process according to claim 3, wherein said size exclusion chromatography process of step (c) is selected from the group consisting of gel filtration chromatography and gel permeation chromatography, and said plant extract fractions are concentrated in step (e) by a method selected from the group consisting of reverse osmosis, microfiltration, ultrafiltration, nanofiltration, hollow fiber diafiltration and any combination thereof.

30. The process according to claim 29, wherein said size exclusion chromatography process is gel filtration chromatography and uses a column packing selected from the group consisting of neutral cross-linked dextran and polyacrylamide gels.

31. The process according to claim 30, wherein said column packing is a neutral cross-linked dextran having a nominal exclusion limit of 700 Daltons.

32. The process according to claim 1, wherein said size exclusion process of step (c) uses an eluant selected from the group consisting of water, ethanol, and any combination thereof.

33. The process according to claim 32, wherein said eluant is water.

34. The process according to claim 29, wherein said size exclusion process is performed at a temperature in the range of about 20°

C. to about 27°

C.

35. The process according to claim 1, wherein prior to isolating said fraction of plant extract by said size exclusion process of step (c), said extract is adjusted to ambient temperature.

40. The process according to claim 1, wherein said concentrated plant extract of step (b) has a total solids content of about 12% to 36% percent by weight.

41. The process according to claim 1, wherein said concentrated plant extract of step (b) has a total solids content of about 18% to 22% percent by weight.

42. The process according to claim 1, wherein prior to step (c), said concentrated plant extract is treated to remove insoluble material contained therein.

43. The process according to claim 42, wherein said treatment of concentrated plant extract for removing insoluble material comprises a sequential chilling treatment.

44. The process according to claim 42, wherein said treatment for removing insoluble material is sequential chilling, centrifugation and filtration.

45. The process according to claim 44, wherein said intermediate molecular weight fractions have a molecular weight in the range from about 1,000 Daltons to about 15,000 Daltons.

46. The process according to claim 45, wherein said intermediate molecular weight fractions have a molecular weight in the range from about 1,500 Daltons to about 5,000 Daltons.

47. The process according to claim 42, wherein said treatment of concentrated plant extract for removing insoluble material comprises a precipitation and decantation treatment.

48. The process according to claim 42, wherein said treatment of concentrated plant extract for removing insoluble material comprises a centrifugation treatment.

49. The process according to claim 42, wherein said treatment of concentrated plant extract for removing insoluble material comprises a centrifugation and filtration treatment.

50. The process according to claim 42, wherein said treatment of concentrated plant extract for removing insoluble material comprises a filtration treatment.

51. The process according to claim 30, wherein said column packing is a polyacrylamide gel having a molecular weight fractionation range/nominal exclusion limit of about 100 to about 1,800 Daltons.

52. The process according to claim 1, wherein said intermediate molecular weight fractions have a molecular weight in the range from about 1,000 Daltons to about 15,000 Daltons.

53. The process according to claim 52, wherein said intermediate molecular weight fractions have a molecular weight in the range from about 1,500 Daltons to about 5,000 Daltons.

60. The process according to claim 1 or 6, wherein under thermal conditions, the isolated intermediate molecular weight fractions release aroma or flavors.

6. A process for producing and isolating intermediate molecular weight fractions from tobacco extracts, which fractions are stable and non-volatile under ambient temperature conditions and are useful as flavoring agents, comprising:

(a) contacting tobacco with a solvent to produce a crude tobacco extract;

(b) concentrating said crude tobacco extract;

(c) subjecting said concentrated crude tobacco extract to a size exclusion process wherein said concentrated crude tobacco extract is fractionated into a series of fractionated tobacco extracts including said intermediate molecular weight fractions;

(d) isolating said fractionated tobacco extracts into fractions including said intermediate molecular weight fractions by using a suitable detection method to detect when said intermediate molecular weight fractions have been eluted from said size exclusion process and to trigger a fraction collection means in order to collect said isolated intermediate molecular weight fractions as subfractions;

(e) concentrating said intermediate molecular weight subfractions; and

(f) optionally contacting said subfractions from step (d) which contain nicotine detectable by suitable nicotine detection means with an ion-exchange material to essentially remove the nicotine.

7. The process according to claim 6, wherein said detection method employs a detection means selected from the group consisting of ultraviolet absorption, infrared absorption, refractive index, light scattering, circular dichroism, pH, and electrochemical conductance.

8. The process according to claim 7, wherein said detection means is a combination of ultraviolet absorption and circular dichroism.

15. The process according to claim 6, wherein said fractioned tobacco extracts contain essentially no nicotine.

36. The process according to claim 6, wherein said ion-exchange material comprises a cation exchange moiety selected from the group consisting of carboxymethyl, sulfopropyl, sulfonic acid or carboxylic acid.

37. The process according to claim 36, wherein said ion-exchange material comprises cross-linked dextran having carboxymethylcellulose substituent groups.

38. The process according to claim 36, wherein said ion-exchange material is used with an eluant selected from the group consisting of monovalent salts and NaOH.

39. The process according to claim 38, wherein said eluant is a 0.1 M NaOH solution.

54. The process according to claim 37, wherein the ion exchange material has a molecular weight fractionation range/nominal exclusion limit of about 1,000 to about 5,000 Daltons.

55. The process according to claim 6, wherein said ion-exchange material comprises cellulose having carboxymethylcellulose substituent groups.

56. The process according to claim 6, wherein said ion-exchange material comprises polystyrene having sulfonic acid substituent groups.

57. The process according to claim 6, wherein said ion-exchange material comprises polystyrene having carboxylic acid substituent groups.

58. The process according to claim 37, wherein said ion-exchange material is a microporous bead-former cation exchanger comprising a cross-linked dextran gel having carboxymethyl substituent groups.

59. A process for producing and isolating intermediate molecular weight fractions from plant extracts, which fractions are stable and non-volatile under ambient temperature conditions and are useful as flavoring agents, comprising:

(a) contacting plant matter with a solvent to produce a crude plant extract;

(b) concentrating said crude plant extract;

(c) subjecting said concentrated crude plant extract to a size exclusion process by fractionating said concentrated crude plant extract into a series of fractionated plant extracts, said extracts falling within a range of intermediate molecular weight fraction of equal to or greater than approximately 600 Daltons and equal to or less than approximately 100,000 Daltons; and

(d) isolating said fractionated plant extracts into fractions including said intermediate molecular weight fractions by using a suitable detection method to detect when said intermediate molecular weight fractions have been eluted from said size exclusion process and to trigger a fraction collection means in order to collect said isolated intermediate molecular weight fractions.