Triterpenoid saponins containing an acetylated branched D-fucosyl residue from Quillaja saponaria Molina

Seven novel saponins were isolated from a bark extract of Quillaja saponaria Molina. the compounds were characterized, using mainly NMR spectroscopy, mass spectrometry and chemical methods, as quillaic acid substituted at C-3 with oligosaccharides consisting of various compositions of D-glucuronic acid D-galactose, D-xylose, and L-rhamnose and at C-28 with complex oligosaccharide structures consisting of various compositions of D-xylose, L-rhamnose, D-apiose and a branched 4-O-acetyl-D-fucose residue.     

Effect of dietary saponins from Quillaja saponaria L. on fatty acid composition and cholesterol content in muscle Longissimus dorsi of lambs

The purpose of this study was to evaluate the effects of increasing levels of saponins from Quillaja saponaria on fatty acid (FA) composition and cholesterol content in muscle Longissimus dorsi of lambs. A total of 24 Barbarine lambs were assigned to four dietary treatments: control diet (C) consisting of oat hay ad libitum and 400 g of concentrate (80% barley, 17.5% soybean meal and 2.5% vitamin and mineral supplement); C diet plus 30 ppm of Q. saponaria L. (QS30); C diet plus 60 ppm of Quillaja (QS60); C diet plus 90 ppm of Quillaja (QS90). Saponin supplementation reduced the concentration of C14:1 cis-9 (P = 0.001) and of its desaturation index (P = 0.002). None of the FA intermediates of ruminal biohydrogenation (BH) was affected by Quillaja saponin supplementation (P > 0.05). The concentration of C20:4n-6 was higher in the meat of animals receiving 60 ppm of Quillaja than C and QS30 groups. Supplementing 60 ppm of Quillaja reduced the ratio between α-linolenic and linoleic acids compared with the C group (P = 0.023). We did not find any significant effect of Quillaja saponins on muscle cholesterol level. Further investigations are necessary to assess the metabolic fate of saponins in the rumen and to understand whether there is an effect of saponin on Δ9-desaturase enzyme activity, ruminal BH and cholesterol metabolism in ruminants. Supplementing up to 90 ppm of Quillaja saponins did not produce detrimental effects on the overall meat FA profile.     

Structural studies of triterpenoid saponins with new acyl components from Quillaja saponaria Molina

Eight new triterpenoid saponins were isolated from a bark extract of Quillaja saponaria Molina by silica and reverse phase chromatography. The saponins were characterized by spectroscopic data and chemical methods as phytolaccagenic acid, 22beta-hydroxy-quillaic acid, and echinocystic acid substituted with different oligosaccharides at C-3 and C-28. The O-4 of the fucosyl residue in the 28-O-oligosaccharide was substituted with either acetyl, (S)-2-methylbutanoyl, or (3S,4S)-3-hydroxy-4-methylhexanoyl groups.     

Effect of Quillaja saponaria saponins and Yucca schidigera plant extract on growth of Escherichia coli

Escherichia coli K-12 was exposed to Quillaja saponaria saponins from various commercial firms (Sigma, Roth and Nor-feed) and to an extract of Yucca schidigera plant powder (DK Sarsaponin 30) at different concentrations (0·05–1·0% w/v). A concentration-dependent response was observed. Quillaja saponaria saponins from Sigma increased growth up to 0·1% (w/v) level, whereas Nor-feed and Roth saponins produced maximum growth at a much higher level (0·5 and 0·75%, w/v, respectively). These results suggest that quillaja saponins from various sources differ in their biological activity, although all three saponins had the same content of vanillin-sulphuric acid reactive moieties. The lyophilized water extract from the DK Sarsaponin powder showed maximum growth at 0·1% (w/v) level. The levels at which maximum growth was observed did not change on subjecting the quillaja or yucca saponins to heat treatment in an autoclave (121 °C for 30 min). All the saponins and the plant extract increased growth of Escherichia coli up to a certain concentration and thereafter decreased growth. In spite of the decreased growth at higher levels of saponins, it was higher compared to the control (without saponin) up to levels of 1% (w/v) for all saponins except Quillaja saponins from Sigma, for which the growth was lower at levels of 0·25% (w/v) and higher. Saponins have the potential to modulate microbial growth in natural and artificial fermenters.     

Synergistic interactions of saponins and monoterpenes in HeLa cells, Cos7 cells and in erythrocytes

In phytomedicine complex extracts consisting of phenolics, monoterpenes or saponins are traditionally used. It is often impossible to attribute the biological activity of an extract to one or few compounds. As an explanation of the superior activity of extracts, a synergistic effect of combinations of active compounds has been suggested. Since lipophilic monoterpenes or saponins targeting the biomembrane usually accompany polar polyphenols in phytomedical preparations, we decided to investigate their effect as single substances and in combination to gain further insight into potential synergistic effects of herbal medicine. Combinations of the monoterpenes α-pinene, thymol and menthol with the monodesmosidic saponins digitonin, aescin, glycyrrhizic acid and Quillaja saponin demonstrated strong synergistic activity. The IC(50) of haemolysis was lowered by a factor of 10-100 from 316μg/ml to 2μg/ml for aescin, 157μg/ml to 11μg/ml for Quillaja saponins and 20μg/ml to 3μg/ml for digitonin when combined with thymol. A similar significant synergistic cytotoxicity occurred both in HeLa and Cos7 cells by combining the α-pinene, thymol and menthol with the saponins. The IC(50) of glycyrrhizic acid was lowered by a factor 100 from around 300μg/ml to around 1-10μg/ml and the IC(50) of aescin, digitonin and Quillaja saponins about the factor 10. Monoterpenes and monodesmosidic saponins have a common target, the biomembrane, which is present in all animal, fungal and bacterial cells. Disturbance of membrane fluidity and permeability is the mode of action. This activity is non-specific which makes it extremely difficult for bacteria and fungi to develop resistance. This explains the overall success of these molecules as defence chemicals in the plant kingdom. The synergistic effect of combinations of saponins with monoterpenes opens a complete new field of possible applications in medicine to overcome resistance in multidrug resistant microbial and human cell.     

Steroidal saponins from the tuber of Ophiopogon japonicus

Eight novel steroidal saponins, ophiopogonins H-O (1-8), along with seven known steroidal saponins (9-15) were isolated from the tubers of Ophiopogon japonicus. The structures of these new compounds were determined by detailed spectroscopic analysis, including extensive 1D and 2D NMR data, and the analysis of hydrolytic reaction products. For the first time, rare furostanol saponins with disaccharide moiety linked at position C-26 of the aglycone were reported to be isolated from a natural source.     

Characterization of quillaja bark extracts and evaluation of their purity using liquid chromatography–high resolution mass spectrometry

In the course of development of semi-preparative liquid chromatographic methods for the isolation of individual quillaja saponins from Quillaja saponaria (L.), some commercially available quillaja bark extracts revealed a distinctive and characteristic pattern of additional peaks in the chromatogram that could not be attributed to saponins commonly present in quillaja. To identify these peaks, analytical procedures based on HPLC coupled with high resolution MS detection were optimized which allowed the identification of the additional saponins Mi saponin A, Mi saponin B, Mi saponin C, madhucoside A and madhucoside B. These compounds are known to be the main saponins of the Indian plant Madhuca longifolia (L.). Tandem MS experiments were performed for the unambiguous assignment of the sapogenin. Madhuca saponins yielded a characteristic fragment of protobassic acid, whereas quillaja saponins showed a fragment of quillaic acid as expected. In addition, samples from madhuca seed kernels were analysed to verify the origin of the characteristic chromatographic peak pattern observed frequently in commercially available quillaja bark extracts.     

Characterization of some O-acetylated saponins from Quillaja saponaria Molina

Sixteen saponins were identified from a bark extract of Quillaja saponaria Molina. The compounds were characterized, using NMR spectroscopy, mass spectrometry and monosaccharide analysis, as quillaic acid substituted at C-3 with oligosaccharides consisting of a disaccharide, beta-D-Galp-(1-->2)-beta-D-GlcpA substituted with either D-xylose or L-rhamnose and at C-28 with complex oligosaccharide structures consisting of a disaccharide, alpha-L-Rhap-(1-->2)-4-O-acetyl-beta-D-Fucp, substituted with various amount of D-xylose. D-glucose, D-apiose, and L-rhamnose.     

Membranotropic effect of some triterpene glycosides possessing immunostimulating properties

The peculiarities of the interaction between cell membrane lipids and triterpene glycosides from holothurians Apostichopus japonicus S. and Cucumaria japonica (holotoxin A1 and cucumarioside A2-2, respectively) were studied in comparison with plant saponins from Quillaja saponaria, known as hemolytic, adjuvant, and structure-forming components of immunostimulating complexes. Similar to Quillaja saponins, the sea glycosides, holotoxin A1 and cucumarioside A2-2 were shown to possess a high hemolytic activity (2.6 and 3 microg/ml, respectively) and sterol-depending membranotropic effect mediated by the formation of nonbilayer sterol-lipid-glycoside complexes. At the same time, cucumarioside A2-2 bound exogenic cholesterol only in the presence of membrane lipids, such as phosphatidylcholine or monogalactosyldiacylglycerol, in contrast to Quillaja saponins and holotoxin A1, which bound cholesterol in the molar ratios 1:2 and 1:8, respectively. Moreover, in all cases, tree-component complexes containing cholesterol, lipid, and glycoside exhibited a lower hemolytic activity compared with two-component sterol-glycoside complexes. It was concluded that the hydrophobic medium of cell membranes performs a potentiative role in the effective interaction between triterpene glycosides and "sterol receptors". A method for decreasing the toxicity of membranotropic holothurian glycosides possessing the immunomodulating properties was suggested.     

Regio- and enantioselective oxidation of thiaoleic acids by an algal delta 12-desaturase

Five new spirostanol saponins and a new furostanol saponin were isolated from the fresh bulbs of Lilium candidum. Their structures were elucidated on the basis of spectroscopic analysis, including two-dimensional NMR spectroscopic techniques and the result of acid hydrolysis. The isolated saponins contained a branched triglycoside moiety assigned as O-alpha-L-rhamnopyranosyl-(1-->2)-O-[beta-D-glucopyranosyl-(1-->6)]-beta - D-glucopyranose with the formation of an O-glycosidic linkage to C-3 of the aglycone as the common structural feature. The inhibitory activity of the saponins on Na+/K+ ATPase was evaluated.     

Separation and structural analysis of saponins in a bark extract from Quillaja saponaria Molina.

Six major saponins were isolated from a bark extract from Quillaja saponaria Molina. Solid-phase extraction, followed by a two-step reversed-phase HPLC separation procedure with phosphate and ammonium acetate buffers of different pH values, was used. The compounds were characterised using NMR spectroscopy, mass spectrometry and chemical methods.     

Separation and characterization of saponins with adjuvant activity from Quillaja saponaria Molina cortex

Saponins were purified from Quillaja saponaria Molina bark by silica and reverse phase chromatography. The resulting purified saponins were tested for adjuvant activity in mice. Several distinct saponins, designated QS-7, QS-17, QS-18, and QS-21, were demonstrated to boost antibody levels by 100-fold or more when used in mouse immunizations with the Ag BSA and beef liver cytochrome b5. These purified saponins increased titers in all major IgG subclasses. To determine optimal dose in mice for adjuvant response, QS-7 and QS-21 were tested in a dose-response study in intradermal immunization with BSA in mice; for both of these purified saponins, adjuvant response (determined by stimulation of ELISA titers to BSA) neared maximum at doses of 5 micrograms and was shown to plateau up to the highest dose tested, 80 micrograms. These purified saponins vary considerably in their toxicity, as assessed by lethality in mice; the main component, QS-18, being the most toxic. Saponins QS-7 and QS-21 showed no or very low toxicity in mice, respectively. None of these saponins stimulated production of reaginic antibodies. The monosaccharide composition of these saponins showed similar but distinct compositions with all four containing fucose, xylose, galactose and glucuronic acid. Predominant differences were observed in the quantities of rhamnose, arabinose, and glucose. Monomer m.w. (determined by size exclusion HPLC) were determined to range from 1800 to 2200.     

Isolation and characterization of two saponins from Fagonia indica

Two new bisglycosidic triterpenoid saponins were isolated from the ethanolic extract of the aerial parts of Fagonia indica. They were characterized as 23,28-di-O-β-D-glucopyranosyltaraxer-20-en-28-oic acid and 3β,28-di-O-β-D-glucopyr acid. Furthermore, the conversion of the aglycone to 3β,23-dihydroxy-28,20β-taraxastonolide, nahagenin, during the acidic hydrolysis of the new saponins was studied.     

Industrial uses and sustainable supply ofQuillaja saponaria (Rosaceae) saponins

The bark of the tree Quillaja saponaria, indigenous to Chile, is one of the major sources of industrially used triterpenoid saponins. For decades quillaja extracts have been used as foaming agents in beverages, emulsifiers in foods, wetting agent in photography, etc. Overexploitation of the bark has caused important ecological damage and a shortage of this resource. However, this can still be remedied by using whole quillaja wood (and not just the bark), for the production of saponins. This raw material can be obtained in large quantities from pruning operations, reducing the need to fell trees. This review covers ecological aspects of quillaja exploitation, as well as a discussion of its novel industrial applications.     

Adjuvant activities of saponins from the root of Polygala senega L

Eight pure triterpenoid saponin compounds isolated from the root of Polygala senega L., a plant indigenous to the Canadian prairies, were evaluated for their immunological activity in mouse models. The specific antibody responses of the IgG2a subclass increased significantly when isolated P. senega saponins were used as adjuvants in the immunization of mice with OVA antigen. In addition, increased IL-2 levels were observed in spleen cell cultures from P. senega saponin-immunized mice after in vitro secondary antigen stimulation. The saponins were tested for their toxicity in mice by using a haemolytic activity assay and found to be less toxic than Quillaja saponaria saponins that have long been used as adjuvants in vaccine formulations. This study has shown the potential of P. senega saponins to be considered as a natural source of vaccine adjuvants with biological activity equivalent to the current commercially available saponin adjuvants.     

Immunoadjuvant saponin production in seedlings and micropropagated plants of <Emphasis Type="Italic">Quillaja brasiliensis</Emphasis>

The leaves of Quillaja brasiliensis, a native tree species of southern Brazil, accumulate saponins which have adjuvant activity in vaccines. An efficient micropropagation system for Q. brasiliensis was developed. Plants were obtained from aseptically germinated seeds. High rates of germination were observed on filter paper and aseptic medium substrate (sucrose and light exposure did not affect the germination response). Stem nodal segments were inoculated into Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine. Each explant produced multiple shoots, which elongated satisfactorily on the same medium. Elongated shoots were rooted in a modified MS medium with or without auxins. Best rooting responses were observed in medium containing 10 mg l⁻¹ indole-3-acetic acid under continuous exposure. The rooted explants were acclimatized and successfully transferred to soil, yielding approximately 95% survival after 10 mo. Leaf content of immunoadjuvant saponins in micropropagated plants was not affected by auxin type used for rooting and was comparable to that of field-grown trees. The results indicate that Q. brasiliensis represents an alternative and readily renewable source of biomass for the production of bioactive saponins.     

Variable sensitivity of Trichoderma viride to Medicago sativa saponins

Eight saponins were isolated from alfalfa roots (Medicago sativa). The sensitivity of Trichoderma viride to the saponin varied with the individual saponin isolate. Seven isolates appeared to contain the aglycone, medicagenic acid, and while the other did not, it inhibited the growth of the fungus at higher concentrations than the other isolates. One pair and a triplet of saponins with divergent R_fs evoked near identical biological responses suggesting structural similarity toxic to T. viride.     

Adjuvant and haemolytic activities of 47 saponins derived from medicinal and food plants

Adjuvant and haemolytic activities of 47 saponins purified from medicinal and food plants were examined. The compounds showed various levels of both adjuvant and haemolytic activities. Soyasaponins and lablabosides showed strong adjuvant activity but little haemolytic activity. Jujubosides showed strong adjuvant and haemolytic activities. Escins showed weaker adjuvant activity than the adjuvant-control, but strong haemolytic activity. Comparison of the functional groups of each saponin revealed that the acyl residue in saponin, the aldehyde group at carbon 4 in aglycone, and branched sugar chains attached to aglycone, were not essential for adjuvant activity. Furthermore, saponins with an acyl residue or oxide-ring moiety tended to show haemolytic activity. These results suggest that the adjuvant activity of saponins does not relate with haemolytic activity. It is considered that not only the functional groups themselves, but the overall conformation harmoniously consisting of such functional groups, affects adjuvant activity of saponins.     

Steroidal saponins from the fresh tubers of Ophiopogon japonicus

Eleven novel furostanol saponins, named ophiofurospisides C–E, G–N (1–3, 5–12), one new spirostanol saponin, named ophiopogonin R (13), were isolated from the fresh tubers of Ophiopogon japonicus. Their structures were determined on the basis of spectroscopic techniques (1D and 2D NMR) and HRESIMS. The isolated furostanol saponins possessed two sugar chains located at C-3 and C-26, respectively. Six furostanol saponins (1, 5–9) with disaccharide moiety linked at position C-26 of the aglycone were rare in the plant kingdom.     

Analysis of an immunoadjuvant saponin fraction from Quillaja brasiliensis leaves by electrospray ionization ion trap multiple-stage mass spectrometry

An immunoadjuvant saponin fraction from Quillaja brasiliensis leaves was investigated by direct infusion and liquid chromatography/electrospray ionization ion trap multiple-stage mass spectrometry in negative ion mode (DI-ESI-IT-MSⁿ and LC-ESI-IT-MSⁿ). The aglycone and the sequence of the oligosaccharide residues at C-3 and C-28 were characterized based on MS² and MS³ experiments of the [MH]⁻ ions. According to their [MH]⁻ ions, characteristic product ions and retention times, 27 bidesmosidic saponins, bearing four types of triterpenic aglycones, were tentatively identified.