Main Glycosides Research Articles

Fraxin Promotes the Activation of Nrf2/ARE Pathway via Increasing the Expression of Connexin43 to Ameliorate Diabetic Renal Fibrosis.

Diabetic nephropathy (DN) is quickly becoming the largest cause of end-stage renal disease (ESRD) in diabetic patients, as well as a major source of morbidity and mortality. Our previous studies indicated that the activation of Nrf2/ARE pathway via Connexin43 (Cx43) considerably contribute to the prevention of oxidative stress in the procession of DN. Fraxin (Fr), the main active glycoside of Fraxinus rhynchophylla Hance, has been demonstrated to possess many potential pharmacological activities. Whereas, whether Fr could alleviate renal fibrosis through regulating Cx43 and consequently facilitating the activation of Nrf2/ARE pathway needs further investigation. The in vitro results showed that: 1) Fr increased the expression of antioxidant enzymes including SOD1 and HO-1 to inhibit high glucose (HG)-induced fibronectin (FN) and inflammatory cell adhesion molecule (ICAM-1) overexpression; 2) Fr exerted antioxidant effect through activating the Nrf2/ARE pathway; 3) Fr significantly up-regulated the expression of Cx43 in HG-induced glomerular mesangial cells (GMCs), while the knock down of Cx43 largely impaired the activation of Nrf2/ARE pathway induced by Fr; 4) Fr promoted the activation of Nrf2/ARE pathway via regulating the interaction between Cx43 and AKT. Moreover, in accordance with the results in vitro, elevated levels of Cx43, phosphorylated-AKT, Nrf2 and downstream antioxidant enzymes related to Nrf2 were observed in the kidneys of Fr-treated group compared with model group. Importantly, Fr significantly improved renal dysfunction pathological changes of renal fibrosis in diabetic db/db mice. Collectively, Fr could increase the Cx43-AKT-Nrf2/ARE pathway activation to postpone the diabetic renal fibrosis and the up-regulation of Cx43 is probably a novel mechanism in this process.

СHROMATOGRAPHIC ANALYSIS OF СYCLOSIVERSIOSIDE F

The plants Astragalus pterocephalus growing in Uzbekistan are a source of triterpene glycosides. The main triterpene glycoside, in terms of content, is a cycloartan glycoside – cyclosiversioside F. To obtain an individual biologically active compound cyclosiversioside F with 95% purity, a proposed method involves extraction with methanol, concentration and dilution with an equal volume of water, then followed by a sequential extraction from the aqueous extract with chloroform, ethyl acetate and butanol.
 Then a chromatographic separation of the purified amount of extractives on a column with silica gel, isolation of the substance and precipitation from a solvent system must be performed, followed by recrystallization and drying. The optimal conditions for the isolation and separation of the amount of extractive substances have been developed in order to obtain an individual glycoside.
 Cyclosiversioside F was authenticated by TLC in comparison with an authentic sample. Quantitative analysis of the glycoside was carried out by HPLC. The purity of cyclosiversioside F was confirmed by taking 1H and 13C NMR spectra.

A case of intoxication with tea made from Digitalis purpurea.

We present the case of a 34-year-old woman with recurrent depressive disorder who ingested purple foxglove with suicidal intent. She bought a foxglove plant (Digitalis purpurea) over the internet and used all of its leaves to make a tea that she then drank over a period of a few hours. Seventeen hours later, she developed abdominal pain, emesis and bradycardia and was admitted via the emergency department to the intensive care unit for further treatment and monitoring. The plasma digoxin concentration measured 3.53 nmol/l (therapeutic reference range 0.77–1.50 nmol/l) 21 hours after ingestion of the tea. She remained heamodynamically and neurologically stable, was treated with antiemetics and simple analgesia and did not require digoxin-specific antibodies. Despite normal renal function, her plasma digoxin half-life was prolonged (estimated 76 h), reflecting the long half-life of the parent compound digitoxin which is the main cardiac glycoside in Digitalis purpurea. She was transferred to psychiatric care 48 h after admission. In this report, we compare this case to other similar cases, which to date have only been rarely reported in the literature.

Chemical composition of dried Stevia rebaudiana Bertoni leaves and effect of ultrasound-assisted extraction on total steviosides content in extract

Summary Introduction: Steviol stevioside, which has been used in the production of food products as a low-calorie sweeten, is one of the main glycoside groups in the leaves of the Stevia plants. It is useful for human health. Objective: The main objection of the present study was to find out some major chemical compositions of the dried Stevia rebaudiana Bertoni leaves and an effective, affordable, and environmentally friendly method to reach the high extraction yield of total steviosides from them. Therefore, a novel extraction, ultrasound-assisted extraction (UAE), was carried out to extract total steviosides from dried leaves of S. rebaudiana plant with ethanol of 70% (v/v) as a solvent. Methods: Some major chemical compositions of the dried S. rebaudiana leaves were analyzed according to the AOAC (Association of Official Analytical Chemists) and total steviosides content (TSC) was measured by anthrone-sulphuric acid colorimetric assay with main influencing factors including material/solvent ratio, extraction temperature, and extraction time. Results: The results referred that some chemical compounds such as protein, lipid, fibre, sugar, etc. existed in this material. The best extraction conditions were the sample/solvent ratio of 1:100 (g/ml), extraction temperature of 75oC, and extraction time of 30 min. Conclusion: The highest amount of total steviosides content of 8.894 % was obtained at the optimal extraction condition. Consequently, these results demonstrated that the parameters of UAE were applied successfully for producing total glycosides.

An in vitro Evaluation of Apigenin and Apigenin-7-O-glucoside Against HeLa Human Cervical Cancer Cell Line

pigenin (API) is a phytocompound belonging to the subclass of flavones that can be found in both functional foods as well as medicinal plants. Recent studies have assigned API antioxidant, anti-inflammatory, anti-spasmodic, anti-viral, anti-thrombotic, anti-angiogenic and chemopreventive potential in vitro on various cell lines and/or in experimental animal models. Apigenin-7-O-glucoside (API-7) is one of its main glycosides and can be commonly found in chamomile flowers, parsley, celery. The aim of this study was to evaluate the in vitro anti-proliferative and pro-apoptotic effects of API and its glycoside (apigenin-7-O-glucoside) against HeLa human cervical cancer cells. Results have shown that in the set experimental conditions both the aglycone as well as the heteroside elicit antiproliferative and pro-apoptotic potential against the screened cell line, the aglycone being more active than the heteroside.

Shoot aqueous extract of Manihot esculenta Crantz (cassava) acts as a protective agent against paracetamol-induced liver injury

Manihot esculenta Crantz (Euphorbiaceae), known as cassava, is a widely cultivated plant, considered one of the main sources of food in the tropical and subtropical regions of Africa and Asia. The aim of the present study was the evaluation of the antioxidant and anti-inflammatory effects of cassava shoot aqueous extract (CSAE) on liver injury induced by paracetamol and investigation of its effect on hyperhomocysteinemia. CSAE was administered to male albino rats classified into seven groups: control, treated, and prophylactic groups. A significant reduction in liver enzymes, malondialdehyde, and homocysteine were observed when compared to the paracetamol group, together with an increase in paraoxonase-1. Histopathological and histochemical results indicated that CSAE effectively ameliorate these parameters. Two main flavonol glycosides, quercetin 3-O-rutinoside and kaempferol 3-O-rutinoside, in addition to a minor myricetin 3-O-rutinoside, were identified in CSAE. CSAE showed a therapeutic potential against paracetamol-induced liver injury probably through antioxidant activity of its flavonol glycosides.

Phytochemical profile of the aerial parts of Rehmannia glutinosa liboschitz var. purpurea Makino

Background: The roots of Rehmannia glutinosa (RG) or Rehmanniae Radix are a well-known medicinal material in the Oriental medicine, and its phytochemical profile has been extensively studied with more than 100 individual compounds from Rehmannia species. In contrast, bioactive components of the aerial part of the title plant are largely unknown as only several compounds reported up to date. Objective: The objective was to study on chemical constituents of the aerial parts of the title plant and evaluate the aerial parts as a supplementary source for Rehmanniae radix. Materials and Methods: Solvent extraction, partition, and column chromatography was used to separate individual compounds; spectroscopic data including nuclear magnetic resonance and mass spectrometry were analyzed to determine the chemical structure of the isolates. Results: Eight compounds including five ursane-type triterpenoids for the first time from RG (ursolic acid [1], pomolic acid [2], 2β-hydroxypomolic acid [3], asiatic acid [4] and 7β,24-dihydroxy ursolic acid [5]) and three main glycosides (ajugol [6], aucubin [8], and acteoside [7]) were characterized from the aerial parts of the title plant. Their structures were identified on the basis of spectroscopic data and comparison with those reported in the literature. Conclusion: The current study reveals various ursane triterpenes in the organic portion beside the main hydrophilic glycosides in the RG aerial parts. The occurrence of various ursane triterpenes contributed in part to phytochemical database and evidence of the biological activity associated with potential in use as a medicinal material of the RG leaves.

Design and construction of novel assemblies for monosaccharides and glycosides detection

A series of new cationic pyridine analogues modified boronic acid were synthesized and established fluorescence probes with dye 8-hydroxypyrene- 1,3,6-trisulfonic acid trisodium salt (HPTS). This kind cationic pyridine analogues as quenchers/connectors in two-components probes displayed higher sensitivity and selectivity for monosaccharides than that of boronic acid functionalized benzyl viologens (BBVs) as quenchers. To investigate the effects of the number of positive charges and boronic acid of these quenchers/connectors, a series of new assemblies comprised of HPTS dye as fluorophor and pyridine analogues as quenchers/connectors were established and employed to detect monosaccharides, respectively. These results displayed that TCTB with three positive charges and three boronic acid groups could interact with anionic dye HPTS forming the most sensitive two-component sensor for monosaccharides. Furthermore, TCTB/HPTS assembly may be also employed to detect saccharides in the blood of diabetes patient. Notably, the assembly could distinguish the three main glycosides of the herb forsythia and had a high selectivity for forsythoside A. The work not only provided high effective two-component probes but also exploited a new mode for detecting and distinguishing main ingredients of Chinese herbal medicine.

FLAVONOLGLYCOSIDE OF SOME SIBERIAN SPECIES OF THE SECTION XIPHIDIUM BUNGE OF THE GENUS ASTRAGALUS L.

For the first time, the composition and content of flavonolglycosides was studied by high-performance liquid chromatography (HPLC) and a chromatograms of extracts of leaves of plants of the species of the Xiphidium Bunge section of the genus Astragalus L. Siberia was compared. The leaves of five species were investigated: A. ionae Palibin, A. lenensis Shemetova, Schaulo et Lomon., A. macroceras C. A. Meyer, A. palibinii Polozhij, A. stenoceras C. A. Meyer. There were found 9 main components, among them flavonols isoqurcitrin, rutin and astragalin have been identified. Glycosides G1, G2, G6 and isoquercitrin was found in all species, astragalin there is only in A. macroceras. Rutin not detected in A. stenoceras. The amount of the main glycosides of flavonoids in the leaves of plants in the species A. ionae, A. lenensis and A. palibinii varies between 0.26–0.70%, in A. macroceras – 0.37–0.59%, in A. stenoceras – 0.20% by weight of the absolutely dry raw material. The composition of the components on chromatograms confirms the taxonomic similarity between A. ionae–A. lenensis–A. palibinii. The chromatograms of A. macroceras and A. stenoceras have a large number of common components, however, composition the most abundant is noted in the A. macroceras.

Investigation on some main glycosides content of Stevia rebaudian B. under different concentrations of commercial and synthesized silver nanoparticles

Stevia rebaudiana B. extract as a low-calorie sweetener has gained worldwide approval. Synthesized nanoparticles by plant have been used in different programs such as medical, pharmaceutical and agricultural fields. Here, we report the increase of the main glycosides of stevia plant with silver nanoparticles (AgNPs) treatment. This fact has stimulated research on the analysis and comparison of different concentrations of commercial and synthesized AgNPs on the glycosides of stevia plant by highperformance liquid chromatography (HPLC). After AgNPs synthesis, spectrum absorption of extracts was analyzed in different ranges from 300 to 700 nm and the maximum peak was observed at 420 nm. Transmission electron microscopy (TEM) micrograph of synthesized AgNPs from Stevia rebaudiana extract showed spherical shape with the size of 25 nm. A comparison between quantities of glycoside by HPLC under both treatments of AgNP exhibited that synthesized AgNPs were more effective than commercial AgNPs on stevioside and then rebaudioside. On the other hand, results showed that increasing AgNP concentration in both treatments will lead to glycoside content enhancement. Overall, significant changes were observed in the glycosides in plants under synthesized and commercial AgNP treatment that could be used to create plants with higher quality of glycosides for pharmaceutical approaches.

Antitumor Perspectives of Oleuropein and Its Metabolite Hydroxytyrosol: Recent Updates.

Olive fruit is a significant and promising source of potential bioactive compounds such as oleuropein and hydroxytyrosol. Oleuropein is the ester of elenolic acid and 3,4-dihydroxyphenyl ethanol (HT). It is the main glycoside in olives, the degradation of which results in the formation of hydroxytyrosol in olive oil. Both plays a significant role in the reduction of coronary heart diseases and a certain type of cancers. Both olive oil phenols have an effective role counter to cell proliferation, cell growth, migration, invasion, and angiogenesis. They down regulate the expression of BCL-2 and COX-2 proteins, and reduced DNA damage. Hydroxytyrosol and oleuropein inhibited the multiple stages in colon carcinogenesis; initiation, promotion, and metastasis. They also provide protection against various human cancers including colorectal, skin, breast, thyroid, digestive, lung, brain, blood, and cervical. This review article discusses the anticancer perspectives and mechanisms of oleuropein and hydroxytyrosol in cell cultures and animal and human studies.

Simultaneous quantification of six phenylethanoid glycosides in some Turkish Scutellaria species by a new HPLC-DAD method

A new HPLC-DAD method was developed and validated for simultaneous determination of six main phenylethanoid glycosides (calceolarioside D, neocalceolarioside D, verbascoside, isoverbascoside, leucoseptoside A and martynoside) in the aerial parts of four Scutellaria L. taxa from flora of Turkey. All standard compounds showed a good linearity (R2 > 0.999) in a relatively wide concentration range (1–120 μg/mL). The LOD of the compounds was in the range of 0.104–1.295 μg/mL and the LOQ was in the range of 0.450–2.536 μg/mL. The recoveries of the selected compounds were calculated in the range of 97.46–117.85%. The amounts of the phenylethanoid glycosides showed variation in the extracts. The developed method was found to be accurate, precise and reproducible, and successfully applied to identify and quantify the phenylethanoid glycoside composition of Scutellaria species.

New seminal variety of Stevia rebaudiana: Obtaining fractions with high antioxidant potential of leaves.

The aim of this study was to determine the composition and antioxidant potential of leaves of a new variety of Stevia rebaudiana (Stevia UEM-13). Stevia leaves of UEM-13 contain rebaudioside A as the main glycoside, while most wild Stevia plants contain stevioside. Furthermore can be multiplied by seed, which reduces the cost of plant culture techniques as other clonal varieties are multiplied by buds, requiring sophisticated and expensive seedling production systems. Ethanol and methanol were used in the extraction to determine the bioactive compounds. The methanolic extract was fractionated sequentially with hexane, chloroform, ethyl acetate and isobutanol, and the highest concentration of phenolic compounds and flavonoids was obtained in the ethyl acetate fraction (524.20 mg galic acid equivalent/g; 380.62 µg quercetin equivalent/g). The glycoside content varied greatly among the fractions (0.5% - 65.3%). Higher antioxidant potential was found in the methanol extract and the ethyl acetate fraction with 93.5% and 97.32%, respectively. In addition to being an excellent source for obtaining of extracts rich in glycoside, this new variety can also be used as raw material for the production of extracts or fractions with a significant amount of antioxidant activity and potential to be used as additives in food.

Classification of <i>Stevia rebaudiana</i> Using Near Infrared Spectroscopy and Multivariate Data Analysis

Stevia rebaudiana leaf contains stevioside and rebaudioside A as main diterpene glycosides. These compounds are used as natural sweetener and potentially as drug candidate of diabetes type 2. Rapid and nondestructive method for S. rebaudiana leaves (n = 23) classification based on geographical area and main diterpene glycosides content was carried out using near infrared spectroscopy combined with multivariate data analysis. Linear discriminant analysis (LDA) was applied to discriminate S. rebaudiana leaves based on geographical area. Principal component analysis (PCA) was established to classify S. rebaudiana leaves based on main diterpene glycosides content. HPLC analysis was used as reference data to divide PCA result into groups. LDA model correctly classified 95% of the S. rebaudiana leaves based on geographical area. PCA model correctly classified 95% and 90% of S. rebaudiana leaves based on rebaudioside A and stevioside content, respectively. The classification method using near infrared spectroscopy combined with multivariate data analysis demonstrate potential use of the classification method established as quality control technique of S. rebaudiana leaves.

Identification of differentially expressed genes from Trichoderma atroviride strain SS003 in the presence of cell wall of Cronartium ribicola

The fungal genus Trichoderma has been extensively studied due to its role in the mycoparasitism, and thus developed as biocontrol agent against various plant pathogens. Although the mycoparasitic processes of several Trichoderma species have already been well understood, the information about the mycoparasitic mechanisms of Trichoderma strains resulted from different growth conditions or interacting with different phytopathogens is still limited. In this study, we utilized transcriptome sequencing to identify the differentially expressed genes (DEGs) at 0, 24, 72 and 120 h from T. atroviride strain SS003, growing on an induced-medium with cell walls of Pinus armandii pathogen Cronartium ribicola (CRCW). In total, 86,155,316 reads were obtained with 43,077,658 clean reads. Further, 10,422 genes were identified from four transcriptomes and accounted for 93.89% of annotated genes in T. atroviride IMI 206040 genome, reflecting high-quality sequencing and assembly. In each pairwise comparison, a large number of DEGs were identified with different numbers of genes for up- and down-regulation, respectively. In the presence of CRCW, expression of two main glycoside hydrolase gene families (i.e. chitinase and glucosidase) was induced. Most of 14 secreted enzymes by quantitative real time PCR (qPCR) analysis exhibited a consistent expression pattern with that by RNA-Seq data. This comparative study leads to the identification of phase-specific genes in the interactions of T. atroviride SS003 with C. ribicola, and provides potential molecular targets for improved biocontrol strategies.

Quantitative analysis of uridine diphosphate glycosyltransferase UGT85C2, UGT74G1 and UGT76G1 genes expression in Stevia rebaudiana under different irrigations

Stevia rebaudiana Bertoni is known to the scientific world for its sweetness and steviol glycosides. Steviol glycosides are the secondary metabolites responsible for the sweetness of stevia and have many medical applications. They are synthesized by a biosynthesis pathway operating in the leaves. Stevioside (Stev) and rebaudioside A (Reb A) are two main glycosides compounds of Stevia. Translation products of three Uridine diphosphate glycosyltransferase candidate genes, UGT85C2, UGT74G1 and UGT76G1, were found to have an activity towards the synthesis of Stev and Reb A. In this study, a experiment was designed to measure the expression level of three candidate genes contributing to the biosynthesis of steviol glycosides under three different irrigation intervals using quantitative real-time PCR analysis technique. Irrigation was done at 3, 6 and 9 days irrigation intervals, based on soil moisture content 90, 75 and 60% of field capacity (FC) respectively. The results showed that soil water depletion up to 75% FC lead to significant increase in expression of UGT85C2 and UGT74G1 genes, whereas a significant growth in expression of UGT76G1 gene occurred at 60% FC. Measurement of Reb A contents in treated leaves showed that a significant correlation between UGT76G1 transcription and Reb A accumulation under 60% FC. Also, the comparison of expression patterns of the three target UGT transcripts at 75 and 60% FC showed a highly significant difference in UGT74G1 transcripts in comparison of UGT85C2 and UGT76G1 transcripts. These results indicated that the highest value of Reb A/ Stev ratio as sweetness quality index likely belongs to 60% FC and suggested 9 days irrigation intervals to improve the quantity and quality of glycoside profile by up-regulation and down-regulation of mentioned genes.

Oleuropein Protects Cardiomyocyte against Apoptosis via Activating the Reperfusion Injury Salvage Kinase Pathway In Vitro.

Oleuropein, the main glycoside present in olives, has been reported to have cardioprotective effect, but the exact mechanism has not been clearly elucidated. This study attempted to clarify the cardioprotective effect of oleuropein against simulated ischemia/reperfusion- (SI/R-) induced cardiomyocyte injury in vitro and further explore the underlying mechanism. Here we confirmed that oleuropein reduced the cell injury in neonatal rat cardiomyocyte induced by SI/R evidenced by decreasing MTT dye reduction and LDH activity in the culture medium. Meanwhile, the compound also inhibited reactive oxygen species excessive generation and stabilized mitochondrial membrane potential after SI/R. The flow cytometry assessment results indicated the inhibition of cellular apoptosis with oleuropein treatment. Furthermore, western blot analysis showed that oleuropein attenuated the expression of Cyt-C, c-caspase-3, and c-caspase-9, increased the Bcl-2/Bax ratio, and enhanced the phosphorylation of ERK1/2 and Akt after SI/R. However, the phosphorylation enhancement was partially abolished in the presence of LY294002 (PI3K inhibitor) and U0126 (ERK inhibitor). All these findings indicate that oleuropein has the protective potential against SI/R-induced injury and its protective effect may be partly due to the attenuation of apoptosis via the activation of the PI3K/Akt and ERK1/2 signaling pathways.

Isolation of steviol glycosides by a two-step membrane process operating under sustainable flux

A two-step membrane process was established to isolate the main glycosides (Stevioside and Rebaudioside-A) from stevia plants grown in different regions. The process involves the clarification of aqueous extracts from dried milled stevia leaves with a 0.2μm ceramic membrane followed by ultrafiltration/diafiltration with either a 15 or a 3kDa membrane. During ultrafiltration a range of hydrodynamic operating conditions were evaluated with both membranes (15 and 3kDa) in order to find the best compromise between working under sustainable flux and eliminating most of the contaminants from the extracts. In addition, principles from high performance tangential flow filtration and continuous diafiltration were combined to increase yield recovery and to reduce operation time. Results indicate that aqueous extracts from stevia plants grown in different regions have different contaminants and glycosides concentrations. Therefore, different membrane pore sizes and operating conditions are needed to obtain high sustainable flux values (≥21kg/m2h) during isolation of glycosides. Moreover, the experimental approach used in the concentration/diafiltration process allowed not only for removal of most of the contaminants, but also for an increase in the sweetener yield and a 50% reduction of process time while keeping similar properties of purified extracts, despite their initial composition.

Green extraction of glycosides from Stevia rebaudiana (Bert.) with low solvent consumption: A desirability approach

The sweet flavor of Stevia rebaudiana (Bert.) leaf extract is well known and has raised the interest of huge food companies due to its natural bid. The extraction of their main glycosides stevioside and rebaudiosideA is an important step on the preparation of final Stevia granules. The aim of the work reported here was to study and optimize the dynamic maceration of Stevia leaves using water and ethanol as green solvents. Forinstance, a fractional factorial design was chosen to evaluate the individual effects of the drug powder size, weight ratio of drug to solvent, temperature, agitation, and time on the yield of these glycosides. The glycosides were quantified by high pressure liquid chromatography. An exhaustive extraction by successive maceration steps showed that ethanol 70% was superior to water and ethanol 90% for stevioside and rebaudioside extraction. The liquid extract composition in dry basis and the yield of stevioside and rebaudioside A were significantly affected by the drug to solvent weight ratio, showing that larger volumes of solvent should be used. Furthermore, increasing solvent volume favors the extraction of the stevioside by a twofold factor as compared to rebaudioside A. Among the other factors, only drug powder size affected the yieldof rebaudiosideA significantly. The optimal solution for S. rebaudiana leaves dynamic extraction estimated by desirability functions methodology led to a condition which allows obtaining extraction yields of 2.31 and 1.24% for stevioside and rebaudioside A and their concentrations in dried extract corresponding to 8.38 and 4.51%, respectively. These high yields were obtained with drug to solvent ratio (1:10, w/w) much higher thanprevious works, thus resulting in a more sustainable and green process.

Green extraction of glycosides from Stevia rebaudiana (Bert.) with low solvent consumption: A desirability approach

The sweet flavor of Stevia rebaudiana (Bert.) leaf extract is well known and has raised the interest of huge food companies due to its natural bid. The extraction of their main glycosides stevioside and rebaudioside A is an important step on the preparation of final Stevia granules. The aim of the work reported here was to study and optimize the dynamic maceration of Stevia leaves using water and ethanol as green solvents. For instance, a fractional factorial design was chosen to evaluate the individual effects of the drug powder size, weight ratio of drug to solvent, temperature, agitation, and time on the yield of these glycosides. The glycosides were quantified by high pressure liquid chromatography. An exhaustive extraction by successive maceration steps showed that ethanol 70% was superior to water and ethanol 90% for stevioside and rebaudioside extraction. The liquid extract composition in dry basis and the yield of stevioside and rebaudioside A were significantly affected by the drug to solvent weight ratio, showing that larger volumes of solvent should be used. Furthermore, increasing solvent volume favors the extraction of the stevioside by a twofold factor as compared to rebaudioside A. Among the other factors, only drug powder size affected the yield of rebaudioside A significantly. The optimal solution for S. rebaudiana leaves dynamic extraction estimated by desirability functions methodology led to a condition which allows obtaining extraction yields of 2.31 and 1.24% for stevioside and rebaudioside A and their concentrations in dried extract corresponding to 8.38 and 4.51%, respectively. These high yields were obtained with drug to solvent ratio (1:10, w/w) much higher than previous works, thus resulting in a more sustainable and green process.