Methods to isolate anti-microbials from fruit or seed extracts

Abstract

The invention provides a method of isolating one or more compounds present in fruit juice or seed extracts that have anti-microbial activity.

What is claimed is: 
     
         1 . A method to isolate compounds having anti-microbial activity, comprising:
 a) providing a fruit or fruit seed extract dissolved or suspended in water, alcohol or in a water alcohol mixture;   b) separating the extract on a C8 column into a plurality of fractions; and   c) isolating at least one fraction that is at least 80% by weight of a compound or a plurality of compounds with anti-microbial activity.   
     
     
         2 . The method of  claim 1  wherein the at least one fraction comprises about 1 mg to about 100 mg of the compound. 
     
     
         3 . The method of  claim 1  further comprising subjecting the at least one fraction to rotavapping at 40° C. 
     
     
         4 . The method of  claim 1  further comprising lyophilizing the at least one fraction. 
     
     
         5 . The method of  claim 1  wherein the extract is a filtered extract. 
     
     
         6 . The method of  claim 1  wherein prior to separation on the C8 column, the extract was subjected to separation on a C18 solid phase extraction column and ion exchange chromatography column. 
     
     
         7 . The method of  claim 1  wherein high pressure liquid chromatography is employed to separate the extract. 
     
     
         8 . The method of  claim 1  wherein the at least one fraction has at least about 95% of the compound, 
     
     
         9 . The method of  claim 1  wherein the fruit is a cranberry. 
     
     
         10 . The method of  claim 1  wherein the seed extract is a cranberry seed extract. 
     
     
         11 . The method of  claim 1  wherein a plurality of chromatographic separation techniques are employed to isolate the fraction having the substantially pure compound. 
     
     
         12 . The method of claim  111  wherein the separation includes solid phase exchange, ion exchange chromatography, high performance liquid chromatography or simulated moving bed chromatography. 
     
     
         13 . The method of  claim 1  wherein at least one compound is a flavonol, proanthocyanin, diterpenoid epoxide, or phenylpropanoid. 
     
     
         14 . An antimicrobial compound isolated by:
 a) separating a fruit or fruit seed extract dissolved or suspended in water or in a water alcohol mixture into fractions;   b) separating the fractions with anti-microbial activity into subtractions;   c) isolating a subfraction with anti-microbial activity that is stable to rotavapping at 40° C. and is resistant to protease activity.   
     
     
         15 . The compound of  claim 14  wherein the fruit is a cranberry. 
     
     
         16 . The compound of  claim 14  wherein the seed extract is a cranberry seed extract. 
     
     
         17 . The compound of  claim 14  wherein solid phase exchange, ion exchange chromatography, high performance liquid chromatography or simulated moving bed chromatography, or a combination thereof, are employed for the separation. 
     
     
         18 . The compound of claim  43  wherein a plurality of different separation techniques are employed. 
     
     
         19 . A method to prevent, inhibit or treat a microbial infection comprising:
 administering to a mammal an anti-microbial amount of a composition comprising an isolated fraction of a fruit or fruit seed extract having at least two proanthocyanins, or one or more of a flavonol, diterpenoid epoxide, or phenylpropanoid.   
     
     
         20 . The method of  claim 19  wherein the composition is orally or intravenously administered.