The objective was to identify a line of Stevia sp. with high concentrations of total glycosides and rebaudioside A, from cultivate of Fundo América in Santa Rita Siguas- Arequipa, to propagate it at commercial scale. Participating entities were Fundo América and UC SM, with support from the Program of Science and Technology (FINCyT). Samples of leaves were selected to identify Stevia rebaudiana Bertoni and Stevia puberula hybrids by DNA extraction and amplification with RAPD markers, identifying molecular markers for each one. Three hybrids of Stevia were selected by its higher content of stevioside and rebaudioside A: TU-12, TU-13 and TR-14, using a new method developed in the research through isocratic elution with NH2 columns by HPLC. The proportion found between percentage of stevioside and rebaudioside-A was 7,62%-8,01%, 8,11%-8,56% and 9,33%- 4,21% for the varieties TU-12, TU-13 ...