Identification of Stevioside Using Tissue Culture-Derived Stevia (<i>Stevia rebaudiana</i>) Leaves

Karim, Md. Ziaul; Uesugi, Daisuke; Nakayama, Noriyuki; Hossain, Moazzem; Ishihara, Kohji; Hamada, Hiroki

doi:10.4137/bci.s30378

“…2, Fig. 3), ( [11] methanolic extract of S. rebaudiana leaves contained 9.6% of Stevioside and in another study Stevioside content in leaves was found to be vary from 3.17-9.94% [12]. The results of the previous studies suggest that the synthesis of important secondary metabolites in S. rebaudiana vary with many external and internal factors.…”

Section: Methods Validationmentioning

confidence: 97%

“…According to their results Stevioside was only observed in ex vitro leaves (0.20 g/kg). According toKarim et al, (2015)…”

mentioning

confidence: 99%

Phytochemical analysis of Stevia rebaudiana using HPLC and GC-MS

Gunasena,

Senarath

2023

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0

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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.

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“…According to their results Stevioside was only observed in ex vitro leaves (0.20 g/kg). According toKarim et al, (2015) [11] methanolic extract of S. rebaudiana leaves contained 9.6% of Stevioside and in another study Stevioside content in leaves was found to be vary from 3.17-9.94%[12] Aman et al, (2013) [13]. reported a higher stevioside (82.48 µg/g-DW) content in in vitro derived Stevia plants on a medium containing a combination of BA and Kin.…”

mentioning

confidence: 99%

Comparison of Phytochemicals in naturally grown plants, tissue cultured plants and calli of Stevia rebaudiana using HPLC and GC-MS

Gunasena,

Senarath

2024

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0

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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.

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Influence of calcium and magnesium elimination on plant biomass and secondary metabolites of Stevia rebaudiana Bertoni

Ahmad

¹

,

Deng

²

,

Adeel

³

et al. 2021

Biotech and App Biochem

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This study reports the increment in the secondary metabolites in Stevia rebaudiana plant after exposure to the elimination of Ca and Mg from Murashige and Skoog culture medium. The effect of nutrient stress on regenerants of S. rebaudiana is measured, which reveals significantly enhanced growth parameters, steviol glycosides (SGs) content, and nonenzymatic antioxidants; total phenolic content, total flavonoid content, total antioxidant capacity, total reducing power, and DPPH‐free radical scavenging activity as compared with the control treatment. However, significantly highest amounts are obtained in a medium with only Ca deficiency. The amount of rebaudioside A (Reb A) and stevioside (ST) obtained in the case of Ca‐deficient medium is 4.08 and 0.69%, respectively. It is followed by the results obtained from both Ca‐ and Mg‐deprived medium [Reb A (3.23%) and ST (0.52%)] and the lowest values are obtained from medium lacking Mg only [Reb A (2.60%) and ST (0.40%)]. The most probable adaptation mechanism might be the production of reactive oxygen species by nutrients’ stress, which results in secondary metabolites production as defensive moieties to overcome stress situation. This effective protocol needs to be refined to apply on an industrial scale in bioreactors for increasing quantities of commercially important pharmaceutical compounds.

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Identification of Stevioside Using Tissue Culture-Derived Stevia (Stevia rebaudiana) Leaves

Cited by 4 publications

References 17 publications

Phytochemical analysis of Stevia rebaudiana using HPLC and GC-MS

Phytochemical analysis of Stevia rebaudiana using HPLC and GC-MS

Comparison of Phytochemicals in naturally grown plants, tissue cultured plants and calli of Stevia rebaudiana using HPLC and GC-MS

Influence of calcium and magnesium elimination on plant biomass and secondary metabolites of Stevia rebaudiana Bertoni

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