Stevia rebaudiana Bertoni is a shrub of the Asteraceae family native to South America, especially of Brazil and Paraguay. This plant accumulates a suite of diterpenoid metabolites that are natural sweeteners finding increased use as sugar. In addition, a number of healing properties have been associated to the stevia leaves intake.

The objectives of the present work have been to develop an optimum extraction method of the main steviol glycosides, stevioside and rebaudioside A, from the Stevia leaves studying the influence of the several factors that affect the process. Applying experimental design methodology, parameters such as, the grinding of the leaves, the temperature and time of extraction, the agitation of the process, the quantity of dried leaves and the volume of dissolvent were assessed. In order to eliminate the impurities and to obtain a crystalline white precipitate several procedures were tested.

Moreover, a HILIC chromatographic method to identify and quantify steviol glycosides from stevia leaves were optimized.