Phytochemical analysis of Stevia rebaudiana using HPLC and GC-MS

Abstract Stevia rebaudiana is a medicinal plant rich in important phytochemicals including steviol glycosides. Its’ leaves are the source of non-caloric sweet steviol glycosides and stevioside is the main type. In this study leaves of naturally grown, tissue cultured plants and callus extracts of S. rebaudiana were analyzed using HPLC and GC-MS to determine the possibility of obtaining stevioside and other important phytochemicals through in vitro technology. Phytochemical extraction was done using methanol, evaporated using a rotary evaporator and re-dissolved in acetonitrile: deionized water (3:7) ratio solvent mixture. Stevioside contents were analyzed using HPLC system with C18 column. Methanolic extracts of naturally grown plants, tissue cultured plants and calli were screened using GC-MS and compared. According to HPLC analysis, leaves of tissue cultured plants contained highest amount of stevioside content (11.462 ± 0.03 mg/mL) followed by naturally grown plants and calli. Twenty seven important phytochemicals were identified by GC-MS analysis and out of those fourteen compounds were found to be common in all three samples. Results revealed that in vitro cultures can be used as the better alternative to overcome limitations in conventional propagation and can be used to obtain stevioside and other important phytochemicals instead of natural plants. GC-MS analysis of Stevia extracts revealed the presence of terpenes, fatty acids, aldehydes with biological activities such as antimicrobial, anticancer, anti-inflamatory, antioxidant, antimalarial and etc. Bioactivity of those phytochemicals showed the possible use of Stevia extracts in pharmaceutical, food and perfume industries.