This project consisted of a comparative study of ascorbic acid content, by high performance liquid chromatography (HPLC), of mucilage Aloe vera plant (Aloe barbadensis Miller.) grown in different municipalities in the department of Risalralda (Belen de Umbria, La Florida, Marsella, Mistrato y Santuario). In order to increase knowledge about the composition of Aloe vera grown in Risaralda and contribute to the diversification of uses and promote greater opportunities in the market national and international. Through Oleochemical group of the Technological University of Pereira was implemented two methods for the extraction of ascorbic acid content of mucilage of Aloe vera. On the methodological details each technique used in the laboratory with appropriate references. By the method of extraction of vitamin C with water solution (H2O) – sulfuric acid (H2SO4) pH 2.7 was found recovery rates (87.56 – 119.58) %. The most appropriate operating conditions for the quantification of ascorbic acid by high performance liquid chromatography (HPLC) with ultraviolet detector (UV) were mobile phase: water solution (5% H2O) – sulfuric acid (95% H2SO4) pH 2.7 under isocratic conditions [1], [2]. Stationary phase: Pinnacle DB C18 (3 μm x 100 mm x 3.2 mm) length of maximum absorption: 248 nm and a flow: 0.3 mL / min. The quantification methodology presented good linearity, precision, accuracy and sensitivity to a confidence level of 95%. So was analyzed the variability in results obtained by growing location. It was determined that the highest and lowest value of vitamin C in wet base of Aloe vera was 4,259 mg (± 0,2248) / kg of sample1 y de 1,272 mg (± 0,0550) / kg of sample1, corresponding to crops of Belen de Umbria y Marsella respectively.