High-purity steviol glycosides

Abstract

Methods of preparing highly purified steviol glycosides, particularly Rebaudioside D, are described. The methods include purification from the extraction stage of the Stevia rebaudiana Bertoni plant, purification of steviol glycoside mixtures, Rebaudioside D and Rebaudioside A from a commercial Stevia extract, and purification of Rebaudioside D from remaining solutions obtained after isolation and purification of Rebaudioside A and a high purity mixture of steviol glycosides. The methods are useful for producing high purity Rebaudioside D, Rebaudioside A, and steviol glycoside mixtures. The high purity steviol glycosides are useful as non-caloric sweeteners in edible and chewable compositions such as any beverages, confectioneries, bakery products, cookies, and chewing gums.

1 - 27 . (canceled) 
     
     
         28 . A method for purifying steviol glycosides, comprising the steps of:
 a. providing a solution comprising steviol glycosides;   b. providing a plurality of connected columns packed with an adsorbent resin capable of adsorbing steviol glycosides to create a multi-column system;   c. passing the solution of steviol glycosides through the multi-column system to obtain at least two columns, each column having a different ratio of adsorbed steviol glycosides; and   d. eluting the adsorbed steviol glycosides from at least one column in the multi-column system to obtain an eluted solution comprising steviol glycosides.   
     
     
         29 . The method of  claim 28 , wherein the multi-column system comprises parallel and serial connected columns. 
     
     
         30 . The method of  claim 28 , wherein the multi-column system comprises parallel connected columns. 
     
     
         31 . The method of  claim 28 , wherein the multi-column system comprises serial connected columns. 
     
     
         32 . The method of  claim 28 , wherein the multi-column system comprises 3-15 connected columns. 
     
     
         33 . The method of  claim 28 , wherein a ratio of a volume of a first column of the multi-column system to a volume of a second column of the multi-column is in the range of about 1:1 to about 1:10. 
     
     
         34 . The method of  claim 28 , wherein a ratio of a volume of a last column of the multi-column system to a volume of a penultimate column of the multi-column system is in the range of about 3:1 to about 1:10. 
     
     
         35 . The method of  claim 28 , further comprising the step of removing impurities by washing the multi-column system with a washing solution prior to eluting the adsorbed steviol glycosides. 
     
     
         36 . The method of  claim 35 , wherein the washing solution comprises an aqueous alcohol solution having a water to alcohol ratio ranging from about 99.9:0.1 to about 60:40. 
     
     
         37 . The method of  claim 35 , wherein the washing solution comprises at least one alkali. 
     
     
         38 . The method of  claim 35 , wherein the washing solution comprises at least one acid. 
     
     
         39 . The method of  claim 35 , wherein the washing solution comprises water. 
     
     
         40 . The method of  claim 35 , wherein the adsorbed steviol glycosides are eluted with an aqueous alcohol solution having a water to alcohol ratio ranging from about 60:40 to about 0.1:99.9. 
     
     
         41 . The method of  claim 28 , further comprising the step of drying the eluted solution comprising steviol glycosides to obtain a purified steviol glycoside mixture, wherein the purified steviol glycoside mixture has a purity level of at least about 95% (w/w) on a dry basis. 
     
     
         42 . The method of  claim 28 , further comprising the steps of:
 a. decolorizing and deionizing the eluted solution comprising steviol glycosides to obtain a decolorized, deionized steviol glycoside solution; and   b. drying the decolorized, deionized steviol glycoside solution to obtain a purified steviol glycoside mixture with at least about 95% by weight total steviol glycosides.   
     
     
         43 . The method of  claim 28 , further comprising eluting high Rebaudioside D fractions from the multi-column system to obtain an eluted Rebaudioside D solution. 
     
     
         44 . The method of  claim 28 , wherein the solution of steviol glycosides is obtained by dissolving a composition comprising a steviol glycoside mixture in a solvent. 
     
     
         45 . The method of  claim 28 , wherein the solution of steviol glycosides is prepared by:
 a. providing a biomass of a  Stevia rebaudiana  plant;   b. producing a crude extract by contacting the biomass with water;   c. separating insoluble material from the crude extract to obtain a filtrate containing steviol glycosides; and   d. treating the filtrate to remove high molecular weight compounds and insoluble particles to obtain the solution of steviol glycosides.   
     
     
         46 . The method of  claim 28 , wherein steviol glycosides are selected from the group consisting of Rebaudioside A, Rebaudioside B, Rebaudioside C, Rebaudioside D, Rebaudioside E, Rebaudioside F, Stevioside, Steviolbioside, Dulcoside A, Rubusoside, or other glycoside of steviol and combinations thereof.