Mixture Of Triterpenoid Saponins Research Articles

β-escin activates ALDH and prevents cigarette smoke-induced cell death

BackgroundThe tobacco use is one of the biggest public health threats worldwide. Cigarette smoke contains over 7000 chemicals among other aldehydes, regarded as priority toxicants. β-escin (a mixture of triterpenoid saponins extracted from the Aesculus hippocastanum. L) is a potent activator of aldehyde dehydrogenase (ALDH) – an enzyme catalyzing oxidation of aldehydes to non-toxic carboxylic acids. PurposeThe aim of this study was to evaluate the effect of β-escin on ALDH activity, ALDH isoforms mRNA expression and cytotoxicity in nasal epithelial cells exposed to cigarette smoke extract (CSE). MethodsNasal epithelial cells from healthy non-smokers were treated with β-escin (1 µM) and exposed to 5% CSE. After 6- or 24-hours of stimulation cell viability, DNA damage, ALDH activity and mRNA expression of ALDH isoforms were examined. Results24 h β-escin stimulation revised CSE induced cytotoxicity and DNA damage. Cells cultured with β-escin or exposed to CSE responded with strong increase in ALDH activity. This effect was more pronounced in cultures treated with combination of β-escin and CSE. The strongest stimulatory effect on ALDH isoform mRNA expression was observed in cells cultured simultaneously with β-escin and CSE: at 6 h for ALDH1A1 and ALDH3A1, and at 24 h for ALDH1A3, ALDH3A2, ALDH3B1, and ALDH18A1. Combined β-escin and CSE treatment prevented the CSE-induced inhibition of ALDH2 expression at 24 h. Conclusionsβ-escin is an effective ALDH stimulatory and cytoprotective agent and might be useful in the prevention or supportive treatment of tobacco smoke-related diseases.

Inhibitory effect of β-escin on Zika virus infection through the interruption of viral binding, replication, and stability

β-Escin is a mixture of triterpenoid saponins extracted from horse chestnut seeds that have diverse pharmacological activities, including anti-inflammation, anti-edematous, venotonic, and antiviral effects. In the clinical setting, β-escin is primarily used to treat venous insufficiency and blunt trauma injuries. The anti-Zika virus (ZIKV) activity of β-escin has not been explored. This study investigated the antiviral efficacy of β-escin on ZIKV and dengue virus (DENV) in vitro and then elucidated the underlying mechanism. The inhibitory effects of β-escin on viral RNA synthesis, protein levels, and infection ability were determined using qRT-PCR, Western blotting, and immunofluorescence assays, respectively. To further characterize how β-escin interferes with the viral life cycle, the time-of-addition experiment was performed. An inactivation assay was performed to determine whether β-escin affects ZIKV virion stability. To broaden these findings, the antiviral effects of β-escin on different DENV serotypes were assessed using dose-inhibition and time-of-addition assays. The results showed that β-escin exhibits anti-ZIKV activity by decreasing viral RNA levels, protein expression, progeny yield, and virion stability. β-Escin inhibited ZIKV infection by disrupting viral binding and replication. Furthermore, β-escin demonstrated antiviral activities against four DENV serotypes in a Vero cell model and prophylactic protection against ZIKV and DENV infections.

Standardization of protocol for shoot regeneration in Bacopa monnieri (L.) Pennell using leaf explants

Bacopa monnieri (L.) Pennell commonly called ‘Brahmi’ is used as a medicinal herb since ages. It is known for its revitalizing and nootropic actions in Ayurvedic medicine as it strengthens memory as well as intellect. This activity of the plant has been accredited to a mixture of triterpenoid saponins, sterols and alkaloids. Of these saponins, Bacoside-A and Bacoside-B have been found to be the most important. Present work aimed to establish direct and indirect shoot regeneration protocol in Bacopa monnieri (L.) Pennell using leaf explants. It was found that MS medium supplemented with 1.0 mg/l 2, 4-D and 0.5 mg/l BAP was best for callus formation (53.33%). Leaf explants showed both direct and indirect regeneration and MS medium supplemented with 6.0 μ mol/ was found to be best for shoot regeneration.

Determination and Metabolite Profiling of Mixtures of Triterpenoid Saponins from Seeds of Chilean Quinoa (Chenopodium quinoa) Germplasm

The seed pericarp of Chenopodium quinoa Willd. (quinoa) contains a mixture of triterpenoid saponins conferring undesired organoleptic properties. In this study, we evaluated saponin content and their corresponding sapogenins in 114 different quinoa accessions. Relative saponin content ranged from 0.22 to 15.04 mg/g of seed dry weight among the genotypes studied and the genotype effect was significant (p < 0.001). About 75% of the genotypes could be classified as low-saponin content lines which is promising in view of ongoing plant breeding efforts. In addition to the quantitative determination of saponins, metabolic profiling was performed by LC-FTICR-MS and LC-MS/MS. We obtained highly accurate mass estimation from ion spectra allowing the identification of twelve saponins of the oleanane type. These differ in their aglycons and in the number and type of glycoside units. Interestingly, we identified a saponin compound that, to our knowledge, had not been reported previously. Our study highlights that there is considerable variability concerning saponin content in quinoa, which contributes to the valorization of genetic resources towards the identification of genotypes that could be utilized in current and future quinoa breeding programs.

Investigation into the Protective Effects of Escin on Blood Cells and Cyclophosphamide-Induced Bone Marrow Toxicity in Rats

Cyclophosphamide (CPM) is an extensively used antineoplastic drug that induces toxicity in the health cell because of its metabolites. The main disadvantage of this drug is myelosuppression. Escin (ES) is a natural mixture of triterpenoid saponins that has biological properties such as antioxidant, anti-inflammation, and anti-edematous. The present study aims to investigate the possible effects of ES upon the toxic effects of CPM on blood cells and bone marrow. Spraque Dawley male rats were chosen and categorized into 4 different groups, including the control group, 200 mg/kg CPM, 10 mg/kg ES and ES+CPM. Rats in all groups were discontinued 1 day after injections. After the animals were cut under anesthesia, bone marrow and blood samples were collected. A fall in the number of erythrocyte, hemoglobin, hematocrit, leukocyte (97%), platelets (45%), and bone marrow cells (93.6%) was seen in the CPM group. Conversely, blood cells (erythrocyte, hemoglobin, hematocrit, leukocyte, and platelets), as well as bone marrow cells in ES + CPM had increased by comparison with CPM. Our results show that treating rats with a selected dose range of ES may help decrease the toxic effects of CPM upon blood cells and bone marrow.

The influence of steroidal and triterpenoid saponins on monolayer models of the outer leaflets of human erythrocytes, E. coli and S. cerevisiae cell membranes

The present paper discusses the use of monolayers of lipid mixtures mimicking the composition of biological membranes of bacteria, erythrocyte and yeast in the context of the anti-bacterial, hemolytic and anti-fungal activity of saponins. Saponins are plant-produced glycosidic biosurfactants with either steroidal or triterpenoidal aglycone. In the present study we used digitonin as a representative steroidal saponin (extracted from Digitalis purpurea) and a mixture of triterpenoid saponins from Quillaja saponaria Molina. The effect of saponins was studied first on monolayers consisting of single lipids characteristic for the given type of biological membrane, and then - on model mixed lipid monolayers. Finally, the monolayers were formed from total lipid extracts of natural cell membranes (E. coli and S. cerevisiae) to verify the results obtained in the simplified models. The effect of saponins on monolayers was studied by a combination of surface pressure relaxation, infrared reflection − absorption spectroscopy (IRRAS) and fluorescence microscopy. In line with expectations, sterols (cholesterol and ergosterol) play a major role in the saponin-lipid interactions in monolayers, which may explain especially the hemolytic and antifungal properties of saponins. In contrast, bacterial membranes are devoid of sterols, although the presence of similar compounds may be responsible for their affinity to saponins. Nevertheless, the effect of saponins on bacterial models is less pronounced than for the erythrocyte or fungal ones.

Aescin-Cholesterol Complexes in DMPC Model Membranes: A DSC and Temperature-Dependent Scattering Study

The saponin aescin, a mixture of triterpenoid saponins, is obtained from the seeds of the horse chestnut tree Aesculus hippocastanum. The β-form employed in this study is haemolytically active. The haemolytic activity results from the ability of aescin to form strong complexes with cholesterol in the red blood cell membrane. In this study, we provide a structural analysis on the complex formation of aescin and cholesterol when embedded in a phospholipid model membrane formed by 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). In this work, the temperatures investigated extend from DMPC’s Lβ′ to its Lα phase in dependence of different amounts of the saponin (0–6 mol% for calorimetric and 0–1 mol% for structural analyses) and the steroid (1–10 mol%). At these aescin contents model membranes are conserved in the form of small unilamellar vesicles (SUVs) and major overall structural modifications are avoided. Additionally, interactions between aescin and cholesterol can be studied for both phase states of the lipid, the gel and the fluid state. From calorimetric experiments by differential scanning calorimetry (DSC), it could be shown that both, the steroid and the saponin content, have a significant impact on the cooperative phase transition behaviour of the DMPC molecules. In addition, it becomes clearly visible that the entire phase behaviour is dominated by phase separation which indeed also depends on the complexes formed between aescin and cholesterol. We show by various methods that the addition of cholesterol alters the impact of aescin on structural parameters ranging from the acyl chain correlation to vesicle-vesicle interactions. While the specific saponin-phospholipid interaction is reduced, addition of cholesterol leads to deformation of SUVs. The analyses of the structures formed were performed by wide-angle X-ray scattering (WAXS), small-angle X-ray scattering (SAXS), and small-angle neutron scattering (SANS).

Molecular targets and anti-cancer potential of escin

Escin is a mixture of triterpenoid saponins extracted from the horse chestnut tree, Aesculus hippocastanum. Its potent anti-inflammatory and anti-odematous properties makes it a choice of therapy against chronic venous insufficiency and odema. More recently, escin is being actively investigated for its potential activity against diverse cancers. It exhibits anti-cancer effects in many cancer cell models including lung adenocarcinoma, hepatocellular carcinoma and leukemia. Escin also attenuates tumor growth and metastases in various in vivo models. Importantly, escin augments the effects of existing chemotherapeutic drugs, thereby supporting the role of escin as an adjunct or alternative anti-cancer therapy. The beneficial effects of escin can be attributed to its inhibition of proliferation and induction of cell cycle arrest. By regulating transcription factors/growth factors mediated oncogenic pathways, escin also potentially mitigates chronic inflammatory processes that are linked to cancer survival and resistance. This review provides a comprehensive overview of the current knowledge of escin and its potential as an anti-cancer therapy through its anti-proliferative, pro-apoptotic, and anti-inflammatory effects.

Gymnemic Acid Stimulates In Vitro Splenic Lymphocyte Proliferation

Gymnemic acid is a mixture of triterpenoid saponins of oleanane class, isolated from Gymnema sylvestre Wild R.Br (family: Asclepidaceae), an herbal plant used in traditional medicine to treat diabetes. Effect of gymnemic acid (0.1-20 µg/mL) on in vitro mitogen (concanavalin A and lipopolysaccharide)-induced splenic lymphocyte proliferation was studied using rat as model. Significant (p < 0.05) stimulation of lymphoproliferation was observed in cultures treated with 10 and 20 µg/mL concentration of gymnemic acid in the absence or presence of mitogens. The present study suggests that gymnemic acid has immunomodulatory property, stimulating lymphoid components of immune system, and the traditional knowledge of anti-diabetic property of G. sylvestre is scientifically supplemented with its immunomodulatory properties.

Anti-inflammatory function of ginsenoside Rg1 on alcoholic hepatitis through glucocorticoid receptor related nuclear factor-kappa B pathway

Ethnopharmacological relevanceGinseng is the dried root of Panax ginseng C.A. Mayer. Since ancient times, ginseng has been used as one kind of treatment drug or tonic in China and even other eastern countries like Korea and Japan. Pharmacological active chemical ingredients and its extract of ginseng are a mixture of triterpenoid saponins, collectively called ginsenosides. Among them, ginsenoside Rg1 is the most pharmacological active one. Aim of the studyBased on prior experimental results and the understanding of alcoholic hepatitis, the major aim of this study is to investigate whether Rg1 is beneficial in a rodent model mimic alcoholic hepatic injury associated with binge drinking and explore the underlying possible mechanisms. Materials and methodsC57BL/6 mice were given oral consumption of 6g/kg alcohol 1h after treated with Rg1 (10, 20 and 40mg/kg) or dexamethasone (1mg/kg) for 9 consecutive days. Biochemical analyses were performed and liver fragments were processed for microscopy, immunohistochemistry and western blot analysis. ResultsAccording to our data, Rg1 treatment significantly reversed the high mortality rate induced by alcohol consumption and also alleviated liver impairment as evidenced by the decrease of serum parameters. Meanwhile, histological and ultrastructural analysis of alcoholic groups showed hepatocellular impairment but restored in Rg1-treated groups. Overproductive inflammatory cytokines were also suppressed by Rg1 in alcohol-intoxicated mouse livers. In addition, changes of GR related NF-κB pathway, including phospho-IκB-α, were also modulated to normal levels. ConclusionThis study demonstrates that Rg1 might promote GR mediating the repression of NF-κB and inhibit the inflammatory reactions in alcoholic hepatitis.

Hypotensive and Bradycardic Effects of Quinovic Acid Glycosides from Aspidosperma fendleri in Spontaneously Hypertensive Rats

The Aspidosperma genus (Apocynaceae) represents one of the largest sources of indole alkaloids widely associated with cardiovascular effects. Aspidosperma fendleri, a plant found mainly in Venezuela, has a single phytochemical report in which is revealed the presence of alkaloids in its seeds. This study explored the cardiovascular effects of an ethanolic extract of A. fendleri leaves (EEAF) in spontaneously hypertensive rats (SHR) and its potential bioactive compounds. Using bioguided fractionation, fractions and pure compounds were intravenously administered to SHR and their effects on mean arterial blood pressure (MABP) and heart rate (HR) monitored over time. EEAF induced hypotensive and bradycardic effects as shown by significant reductions in mean arterial blood pressure (MABP) and heart rate (HR), respectively. Bioactivity-guided fractionation led to the isolation of a mixture of two known isomeric triterpenoid glycosides identified by spectral evidence as quinovic acid 3-O-β-rhamnopyranoside and quinovic acid 3-O-β-fucopyranoside. This mixture of triterpenoid saponins induced reductions in MABP and HR similar to those induced by propranolol. Together, these findings indicate that the two quinovic acid glycosides are responsible for the hypotensive and bradycardic effects which suggest their potential use in cardiovascular therapy.

Triterpenoid saponin augmention of saporin-based immunotoxin cytotoxicity for human leukaemia and lymphoma cells is partially immunospecific and target molecule dependent

Context: Saponinum album (SA) is a complex mixture of triterpenoid saponins previously shown to augment the cytotoxicity of the type I ribosome-inactivating protein saporin and an EGF-saporin target toxin that could potentially be used to improve the therapeutic window of targeted toxins.Objective: To investigate the augmentative property of SA on saporin and saporin-based immunotoxins (IT) directed against five different cell surface target molecules on human leukemia and lymphoma cells.Materials and methods: After determining the optimum dose of SA for each cell line, the extent of SA-mediated augmentation was established for saporin and five saporin-based ITs using XTT and an annexin V apoptosis assay. Immunospecificity was investigated using three different blocking assays. Dose-scheduling was also investigated using the XTT assay.Results: Uncorrected SA-mediated augmentation ranged at best from 31.5 million-fold to, at worse, 174-fold. However, when the calculated fold-increases were adjusted for the non-immunospecific effects of SA on an off-target IT, the true augmentative effects of SA were found to be largely non-immunospecific. Antibody blocking studies demonstrated that the augmentative effect of SA was only partially immunospecific. Separate exposure of target cells to IT and SA at different times demonstrated that immunospecific augmentation of IT by SA could be achieved but only if cells were exposed to IT first and SA second.Conclusions: SA significantly, although variably, augments the cytotoxicity of saporin and saporin-based immunotoxins. Concomitant exposure to both IT and SA can result in non-immunospecific cytotoxicity that can be overcome by temporally separating exposure to each.

Cucurbitane glycosides derived from mogroside IIE: structure-taste relationships, antioxidant activity, and acute toxicity.

Mogroside IIE is a bitter triterpenoid saponin which is the main component of unripe Luo Han Guo fruit and a precursor of the commercially available sweetener mogroside V. In this study, we developed an enzymatic glycosyl transfer method, by which bitter mogroside IIE could be converted into a sweet triterpenoid saponin mixture. The reactant concentration, temperature, pH and buffer system were studied. New saponins with the α-glucose group were isolated from the resulting mixtures, and the structures of three components of the extract were determined. The structure-taste relationships of these derivatives were also studied together with those of the natural mogrosides. The number and stereoconfiguration of glucose groups present in the mogroside molecules were found to be the main factor to determine the sweet or bitter taste of a compound. The antioxidant and food safety properties were initially evaluated by their radical scavenging ability and via 7 day mice survival tests, respectively. The results showed that the sweet triterpenoid saponin mixture has the same favorable physiological and safety characteristics as the natural mogrosides.

Synergistic protective effects of escin and low-dose glucocorticoids on blood-retinal barrier breakdown in a rat model of retinal ischemia

Escin, a natural mixture of triterpenoid saponins isolated from the seed of the horse chestnut (Aesculushippocastanum), has been demonstrated to possess glucocorticoid (GC)‑like anti‑edematous and anti‑inflammatory effects. The aim of the present study was to investigate whether escin exhibits synergistic protective effects on blood‑retinal barrier (BRB) breakdown when combined with GCs in a rat model of retinal ischemia. Low concentrations of escin and triamcinolone acetonide (TA) alone did not affect BRB permeability. However, when administered together, low‑dose escin and TA significantly reduced BRB permeability following ischemia. Furthermore, low‑dose escin and TA alone did not affect the expression of occludin in the ischemic retina; however, when administered together, they significantly increased occludin expression in the ganglion cell layer of the ischemic retina. This indicates that escin and GCs have synergistic protective effects on BRB breakdown and the molecular mechanisms may be correlated with the upregulation of occludin. Therefore, the administration of escin may allow a reduction in the dose of GCs for the treatment of macular edema. The combination of escin with GCs is potentially a beneficial treatment method for BRB breakdown and warrants further investigation.

Escin exerts synergistic anti-inflammatory effects with low doses of glucocorticoids in vivo and in vitro

Escin, a natural mixture of triterpenoid saponins isolated from the seed of the horse chestnut (Aesculus hippocastanum), had been demonstrated to possess anti-edematous and anti-inflammatory effects. The present study was designed to investigate whether escin exhibits synergistic anti-inflammatory effects when combined with glucocorticoids. The carrageenan-induced paw edema and pleuritis in bilaterally adrenalectomized rats were used to investigate the anti-inflammatory effects of escin and glucocorticoid alone or combined. The carrageenan-induced paw edema was inhibited only when escin and corticosterone (Cort) were administered together. Co-administration of escin with Cort significantly reduced the volume of exudates and the number of white blood cells of exudates in bilaterally adrenalectomized rats with pleuritis, but treatment with escin or Cort alone at a suboptimal concentration did not show any effect on the pleuritis rats. After the murine macrophagic RAW264.7 cells stimulated by lipopolysaccharide (LPS), they were treated with escin, Cort or escin and Cort. Then nitric oxide (NO), tumor necrosis factor-α (TNF-α) and interleukin 1β (IL-1β) of cell culture supernatants were analyzed. Escin or Cort markedly reduced the content of NO, TNF-α and IL-1β secreted by LPS-stimulated RAW264.7 macrophage cells. The combination of suboptimal concentrations of escin with Cort, which alone could not markedly inhibit the release of inflammatory factors, inhibited the secretion of NO, TNF-α and IL-1β in LPS-stimulated RAW264.7 macrophage cells. The findings suggest escin can synergize with glucocorticoids to enhance their anti-inflammatory effect.

Rapid quantification of 14 saponins of Maesa lanceolata by UPLC-MS/MS

Triterpene saponins are a class of plant natural products with a wide range of bioactivities, and therefore they are a promising research subject. In previous work a triterpenoid saponin mixture was isolated from the leaves of Maesa lanceolata and the compounds were identified [1,2]. These compounds showed virucidal, haemolytic, molluscicidal and anti-angiogenic activity [3,4]. Maesasaponin II displays the highest anti-angiogenic activity, but is only present in very small amounts in the plant. To increase this amount, a platform of combinatorial biosynthesis in the plant was developed. By introducing genes involved in saponin biosynthesis we are attempting to identify new active compounds, and a higher production of the known compounds. In the first phase of the project, only small amounts of transgenic plant material are available. Therefore it is important to use very sensitive analytical methods. For the fast and sensitive analysis of the extracted and purified plant samples, ultra-performance liquid chromatography (UPLC) was coupled to a triple-quad mass spectrometer for MS/MS detection (TQD). The intensity of the signal obtained by fragmentation of the sodium adducts of the saponins was optimized by addition of sodium acetate to the mobile phase. The method was linear over the investigated concentration range with a correlation coefficient higher than 0.99. Furthermore the method was shown to be repeatable and accurate and therefore suitable for screening of the saponin production of plants transformed with genes involved in saponin biosynthesis.

Anticancer activities and the drug targets of botanical saponins from a medicinal herb, gynostemma pentaphyllum

AACR Annual Meeting-- Apr 14-18, 2007; Los Angeles, CA 4985 Triterpenoid saponins are complex molecules produced by most plant species through complex biosynthetic pathways. These plant metabolites play important roles to humans owing to their diverse medicinal effects, including antiinflammatory, antimicrobial, and anticancer activities. In our laboratory, we have been investigating the anticancer activities of a mixture of triterpenoid saponins (designated GpMix) extracted from a medicinal plant, Gynostemma pentaphyllum known as Jiaogulan or southern ginseng in China. We found that the GpMix effectively inhibits the growth of tumor cells. Intriguingly, the inhibitory effects of GpMix require the presence of co-cultivated normal cells. The GpMix is also able to substantially reduce the number of polyps in the Min/+ mice that are predisposed to colon cancer. The potential gene targets of GpMix were investigated. Based on the results obtained from western, northern blotting analysis and protein expression in situ, GpMix does not affect the transforming ability, subcellular localization, or the level of Ras protein. Instead, the Raf-1 protein was found to be totally degraded within 2 days of GpMix treatment. In addition, alteration of phosphorylation of the Erk1/2 genes was detected upon GpMix treatment. Further proteomic analyses revealed that a wide spectrum of glycolytic and energy metabolic gene products were downregulated in cells treated with GpMix. Literatures showed that majority of tumor cells overexpress glycolytic enzymes to compensate the low rate of ATP synthesis. Our findings may provide a conceptually novel approach by which the growth of cancer cells can be suppressed through the reduction of the cellular glycolytic activities.

Determination of saponins in Maesa lanceolata by LC-UV: Development and validation

Triterpene saponins are a class of plant natural products with a wide range of bioactivities, which makes them an interesting research subject. The small tree Maesa lanceolata, growing in African countries, is used in traditional medicine against various diseases. In previous work a triterpenoid saponin mixture was isolated from the leaves of M. lanceolata and the compounds were identified as closely related oleanane type triterpenes [Apers, S., Foriers, A., Sindambiwe, J.B., Vlietinck, A., Pieters, L., 1998. Separation of a triterpenoid saponin mixture from Maesa lanceolata: semi preparative reversed-phase wide pore high performance liquid chromatography with temperature control. J. Pharm. Biomed. Anal. 18, 737; Apers, S., De Bruyne, T.E., Claeys, M., Vlietinck, A.J., Pieters, L.A.C., 1999. New acylated triterpenoid saponins from Maesa lanceolata. Phytochemistry 52, 1121]. The compounds showed virucidal, haemolytic, molluscicidal and antiangiogenic activity [Apers, S., Baronikova, S., Sindambiwe, J.B., Witvrouw, M., De Clercq, E., Vanden Berghe, D., Van Marck, E., Vlietinck, A., Pieters, L., 2001. Antiviral, haemolytic and molluscicidal activities of triterpenoid saponins from Maesa lanceolata: establishment of structure–activity relationships. Planta Med. 67, 528; Apers, S., Bürgermeister, J., Baronikova, S., Vermeulen, P., Paper, D., Van Marck, E., Vlietinck, A.J., Pieters, L.A.C., 2002. Antiangiogenic activity of natural products: in vivo and in vitro test models. J. Pharm. Belg. 57 (Hors-série 1), 47]. Here we report the development of an extraction and quantification method to analyse saponin compounds in roots and leaves of M. lanceolata. After a purification step using C 18 solid phase extraction (SPE) cartridges, the samples were analysed on a LC-UV/MS system. The identification of the peaks from the different saponins was confirmed based on the retention time and mass spectrum. The quantification was performed using the UV signals. The standard oleanolic acid curve was linear over a concentration range of 2.8–140.0 μg/mL. The recovery from the leaves was 94.5%. The precision of the method with respect to time and concentration was acceptable, with relative standard deviation (RSD%) values of 4.9 and 4.3, respectively.

Quantitative method development for measurement of Maesa lanceolata saponins by LC-MS

Triterpene saponins are a class of plant natural products with a wide range of bioactivities, which make them an interesting research subject. The small tree Maesa lanceolata, growing in African countries, is used in traditional medicine against various diseases. In previous work a triterpenoid saponin mixture was isolated from the leaves of Maesa lanceolata and the compounds were identified [1, 2]. The compounds showed virucidal, haemolytic, molluscicidal and anti-angiogenic activity [3, 4]. Here we report the development of a quantitative LC-MS method to analyse saponin compounds in crude root and leaf extracts of Maesa plants. The crude extract is first purified on a C18ec SPE column. Then the compounds are separated on a reversed phase C18 column (Grace Vydac, 3.2mm x 250mm –300Å) with a H2O/ACN (0.06% formic acid) gradient before analysis with mass spectrometry (Bruker Daltonics Esquire 3000 plus). This method will be used to screen plants whether or not induced by methyl-jasmonate, or plants such as Medicago truncatula transformed with genes involved in saponin biosynthesis, for the presence of Maesa saponins.

Rapid analysis of a triterpenoid saponin mixture from plant extracts by electrospray ionization multi-stage tandem mass spectrometry (ESI-MSI).

Rapid analysis of a triterpenoid saponin mixture from plant extracts by electrospray ionization multi-stage tandem mass spectrometry (ESI-MSI).