# US 11,306,114 B2 · Process for separation, isolation and characterization of steviol glycosides

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- Number: US 11,306,114 B2
- Document ID: US11306114B2
- Status: Granted
- Kind: B2
- Application number: 14/624,758
- Filed: 2015-02-18
- Published: 2015-09-17
- Granted: 2022-04-19
- Earliest claimed: 2014-02-18
- Assignees: Heartland Consumer Products, LLC
- Applicants: Heartland Consumer Products, LLC
- Inventors: Steven J. Catani; Juan Navia
- Web page: https://patentgrep.com/patent/US11306114B2

## CPC

- C07H 1/08: from natural products
- A23L 27/33: {containing sugars or derivatives}
- C07H 15/24: Condensed ring systems having three or more rings
- A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs
- C07H: SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS (derivatives of aldonic or saccharic acids C07C, C07D; aldonic acids, saccharic acids C07C59/105, C07C59/285; cyanohydrins C07C255/16; glycals C07D; compounds of unknown constitution C07G; polysaccharides, derivatives thereof C08B; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; sugar industry C13)
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION

## Abstract

A comprehensive process for the separation, isolation and characterization of a combination of two or more steviol glycosides from extract of Stevia rebaudiana plants and their use in sweetening compositions are disclosed. Combinations of two or more steviol glycosides from Stevia rebaudiana are characterized. The combinations of two or more steviol glycosides can be used as sweetness enhancers, flavor enhancers and sweeteners in foods, beverages, cosmetics and pharmaceuticals. A method for isolating combinations of two or more steviol glycosides is also disclosed.

## Claims

### Claim 1

A process for production a steviol glycoside composition comprising:
a) extracting dried Stevia leaves for 2 h at 95° C. to form a crude extract;
b) fractionating the crude extract on a reverse-phase column, wherein fractionation is conducted using a gradient of water and ethanol, wherein said gradient is used starting from 1:0, moving to 29.8:70.2, moving to 23.4:76.6, moving to 0:1, and moving to 1:1 water to ethanol, where the ethanol is food grade ethanol; wherein, fractions containing rebaudioside D, N, M, and A are collected and the water and ethanol removed, to provide a steviol glycoside composition comprising 23% rebaudioside D, 21% rebaudiosides N and M, and 2% rebaudioside A; and wherein,
rebaudiosides D, N, M, and A are obtained in the claimed percentages without the need for crystallization.

### Claim 2 (depends on 1)

The process of claim 1, wherein the Stevia leaves have not been desiccated.

### Claim 3 (depends on 1)

The process of claim 1, wherein the Stevia leaves have been desiccated.

### Claim 4 (depends on 1)

The process of claim 1, wherein the resulting composition further comprises one or more of rebaudiosides I and O.

### Claim 5 (depends on 1)

The process of claim 1, wherein the crude extract is clarified by centrifugation or filtration before fractionation.