Simultaneous determination of several flavonoids and phenolic compounds in nineteen different Cephalaria species by HPLC-MS/MS.

Cephalaria species in Turkey known as "Pelemir" and they have many different biological activities due to their wide range of chemical content. The main goal of this study is determine the flavonoids and phenolic acids in Cephalaria species using high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC-MS/MS). This method was developed for quantitation of reported compounds using reverse phase Fortis C18 (150 × 3 mm × 5 µm) column with a gradient of 0.1% formic acid solution (A) in water and (B) in methanol in ESI mode. The recoveries of developed method were obtained in the range of 90.4-109.4%. Limit of detection (LOD) and limit of quantification (LOQ) values were calculated in the range of 0.3-25.2 ppb and 3.0-102.3 ppb for the reported compounds. Uncertainty evaluations of all compounds were reported. Twenty-five flavonoids, five simple phenolics, two triterpenoids, one iridoid and vitamin C successfully identified and quantified for nineteen Cephalaria species. The main flavonoids in the studied species were determined as cyanidin-3-O-glucoside, luteolin-7-O-glucoside, kaempherol-3-O-glucoside, hyperoside, hesperidin, luteolin and quercetin while salicylic and caffeic acids were detected as major phenolic acids in the analyzed samples. This is the first full characterization study for nineteen Cephalaria species via HPLC-MS/MS.

Increased cytotoxic potential of infrequent triterpenoid saponins of Cephalaria taurica obtained through alkaline hydrolysis.

Phytochemical investigations of the aerial parts of the plant Cephalaria taurica Szabó. (Caprifoliaceae) have resulted in the isolation of nine oleanane-type triterpenoid saponins, of which two still remain undescribed. The structures of tauricosides A and B were characterized based on NMR analysis, HRESIMS spectrometry, and chemical evaluations. The saponins tauricosides A and B have been rarely reported in the literature due to the presence of eight sugar moieties, and this is also the first report of saponins containing eight sugar moieties in the Caprifoliaceae family. The cytotoxic activities of tauricosides A and B, their undescribed prosapogenins, aglycone hederagenin, and n-butanol extract of C. taurica against the cancerous cells A-549, HeLa, PANC-1, and SH-SY5Y and the noncancerous HEK-293 cells were evaluated by the MTT method. Although tauricosides A and B and the crude n-butanol extract did not exhibit any activity at the tested concentrations on all the tested cells, after alkaline hydrolysis, the cytotoxic activity potential of the compounds was obviously improved. The most active compound, obtained after the alkaline hydrolysis of tauricoside B, showed a significant inhibitory effect, which was higher than that of the standard, commercially available drug doxorubicin, on the cancerous A-549, HeLa, PANC-1, and SH-SY5Y cells with IC50 values of 9.04, 8.75, 6.87, and 4.32 µM, respectively. In addition, prosapogenin, obtained after the alkaline hydrolysis of tauricoside A, exhibited considerable cytotoxic activity on the cancerous A-549, HeLa, PANC-1, and SH-SY5Y cells, with IC50 values of 13.19, 10.32, 11.91, and 7.49 µM, respectively. In conclusion, the alkaline hydrolysis of the saponins (tauricosides A and B) obviously improved their cytotoxic activity potential.

Immunomodulatory, hemolytic and cytotoxic activity potentials of triterpenoid saponins from eight Cephalaria species.

BACKGROUND: Saponins isolated from a number of plants possess a broad spectrum of biological and pharmacological activities by using in vitro and in vivo bioassays. The recent investigations and findings in biological activity studies of saponins have mostly focused on immunomodulatory, hemolytic and cytotoxic properties. HYPOTHESIS/PURPOSE: Considering the great potential of saponins as bioactive agents, we investigated the cytotoxic, hemolytic and immunomodulatory activities of nineteen triterpenoid saponins from the aerial parts of eight Cephalaria species from Turkey. STUDY DESIGN: The isolated oleanane type saponins from Cephalaria species were tested for their hemolytic, cytotoxic and immunomodulatory activities through anin vitro cell based assay systems. METHODS: HeLa, A549, and a normal cell line HEK293 were used for testing cytotoxicity using MTT method. Immunomodulatory activity was performed in stimulated whole blood cells by PMA plus ionomycin. Hemolytic activity was assessed on human erythrocytes. RESULTS: Aristatoside C (3) and davisianoside B (16) displayed significant inhibitory effects on cancerous A549 and HeLa cells, and non-cancerous HEK293 cells with IC50 values of 3.52 ± 0.11, 35.69 ± 0.50, 8.96 ± 0.62 µM and 4.08 ± 0.06, 11.74 ± 0.82, 20.43 ± 3.21 µM, respectively. Thirteen saponins out of the nineteen tested increased IL-1ß concentrations considerably, while six compounds changed IL-2 or IFN-γ levels slightly. Almost all of the saponins showed noticeable hemolysis in human erythrocyte cells based on the sugar units. CONCLUSION: In this study, almost all saponins with oleanolic acid as aglycones exhibited significant hemolysis, monodesmosidic saponins with hederagenin as aglycone were the most active compounds against lung cancer cells with greater activity than standard commercial chemotherapy drug doxorubicin and some of the hederagenin type saponins induced remarkable IL-1ß secretion.

Chemical constituents and biological activities of Dianthus elegans var. elegans.

Chemical investigation of the aerial parts of Dianthus elegans var. elegans afforded two previously undescribed saponins, named dianosides M-N (1-2), together with four oleanane-type triterpenoid glycosides (3-6). Their structures were elucidated as 3-O-α-L-arabinofuranosyl-16α-hydroxyolean-12-ene-23α, 28ß-dioic acid (1) and 3-O-α-L-arabinofuranosyl-(1 â†’ 3)-ß-D-glucopyranosyl 16α-hydroxyolean-12-ene-23α-oic acid, 28-O-ß-D-glucopyranosyl-(1 â†’ 6)-ß-D-glycosyl ester (2) by chemical and extensive spectroscopic methods including IR, 1D, 2D NMR and HRESIMS. Both of the saponins were evaluated for their cytotoxicities against HEK-293, A-549 and HeLa human cancer cells using the MTT method. All compounds showed no substantial cytotoxic activity against tested cell lines. However, dianosides M-N and the n-butanol fraction exhibited considerable haemolysis in human erythrocyte cells. The immunomodulatory properties of dianosides M-N were also evaluated in activated whole blood cells by PMA plus ionomycin. Dianosides M-N increased IL-1ß concentration significantly whereas the n-butanol fraction slightly augmented IL-1ß secretion. All compounds did not change IL-2 and IFN-γ levels considerably.

Immunomodulatory, hemolytic properties and cytotoxic activity potent of triterpenoid saponins from Cephalaria balansae.

Phytochemical investigations on n-butanol extract of Cephalaria balansae Raus. (Caprifoliaceae) led to the isolation of four previously undescribed triterpenoid saponins based on hederagenin type aglycone, namely, balansoides A-D, along with ten known compounds. Their structures were proposed based on 1D and 2D NMR spectroscopic data, HRESIMS analysis and chemical evidence as 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin, 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-glucopyranosyl ester, 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-galactopyranosyl-(1 â†’ 6)-ß-D-glucopyranosyl ester and 3-O-α-L-rhamnopyranosyl-(1 â†’ 3)-ß-D-xylopyranosyl-(1 â†’ 3)-α-L-rhamnopyranosyl-(1 â†’ 2)-α-L-arabinopyranosyl hederagenin 28-O-ß-D-glucopyranosyl-(1 â†’ 6)-ß-D-glucopyranosyl ester. The four saponins were evaluated for their potential cytotoxic activity against HEK-293, A-549, HeLa cells and for hemolytic properties on human blood cells. Balansoide A displayed significant inhibitory effects on cancerous A-549 and HeLa cells, and non-cancerous HEK-293 cells with IC50 values of 12, 15 and 8 µM, respectively. Balansoides A-D together with n-butanol extract exhibited considerable hemolysis in human erythrocyte cells. Immunomodulatory properties of balansoides A-D were also evaluated in activated whole blood cells by PMA plus ionomycin. While balansoides A-C increased IL-1ß concentration with values of 1004.47, 991.57 and 966.50 pg/ml, only balansoide B augmented a slight IFN-γ secretion with value of 5219.14 pg/ml. None of the compounds changed IL-2 levels significantly.

Isolation and identification of a new saponin from Cephalaria aytachii.

A new hederagenin-type triterpenoid glycoside (1) named Aytachoside A, along with eight known triterpene glycosides, were isolated from the aerial parts of Cephalaria aytachii Gokturk and Sumbul (Dipsacaceae). The structures of compounds 1-9 were determined by spectroscopic (1D and 2D NMR, HRESIMS) and chemical examinations. The antimicrobial effect of compound 1 was found considerably active against Escherichia coli, Pseudomonas aeruginosa and especially Salmonella typhimurium microorganisms using the MIC method. Although compound 1 was found not to have a remarkable toxic effect at a concentration lower than 300 µg/mL, cytotoxic activity tests demonstrated that prosapogenin 1a exhibits a significant cytotoxic activity against HeLa cell lines using the MTT assay for the first time.

Toxicity of Cephalaria Species and their Individual Constituents against Aedes aegypti.

Crude acetone and ethanol extracts of the aerial parts of 21 Cephalaria species collected from Turkey were investigated for larvicidal and adult topical activity against Aedes aegypti. The ethanol extracts from C. elazigensis var. purpurea, C. anatolica, and C. elmaliensis possessed the highest mortality against first instar Ae. aegypti larvae. Luteolin-7-O-ß-D-glycoside (1), isolated from C. elmaliensis ethanol extract, demonstrated 33% and 53% mortality at 0.1 µg/mL concentration against first instar ORL (susceptible) and PR (pyrethroid resistant) strains, respectively. C. scoparia acetone extract showed 100% mortality against adult Ae. aegypti. From this extract compounds 2-8 were isolated. Compound 2 (isoorientin) possessed the highest toxicity with 31.7% and 65% mortality at a 10 µg/mL concentration against adult ORL and PR strains, respectively. This is the first screening report of potential insecticides from Cephalaria species against the yellow fever mosquito, Ae. aegypti, and the active compounds (1 and 2) could lead to the development of a new class of insecticide.

Albiziaflavane A: a new flavane from Albizia ferruginea (Mimosoideae).

A new polyhydroxylated flavane (1) was identified from Albizia ferruginea (Mimosoideae) along with 4',7-dihydroxyflavan-3,4-diol (2), julibrosides A1-A3 (3-5). The structures of the compounds were established based on their NMR and mass spectrometric data in conjunction with those previously reported in the literature. Compounds 1 and 5 displayed moderate antibacterial activities.

Comparison of antioxidant capacity, protein profile and carbohydrate content of whey protein fractions.

Whey is used as an additive in food industry and a dietary supplement in nutrition. Here we report a comparative analysis of antioxidant potential of whey and its fractions. Fractions were obtained by size exclusion chromatography, before and after enzymatic digestion with pepsin or trypsin. Superoxide radical scavenging, lipid peroxidation inhibition and cupric ion reducing activities of different fractions were checked. Peptides were detected by SDS-PAGE and GC-MS was used to determine carbohydrate content of the fractions. All samples showed antioxidant activity and the second fraction of the trypsin hydrolysate showed the highest superoxide radical scavenging activity. CUPRAC value of this fraction was two-times higher than that of whey filtrate. The first fraction of the pepsin hydrolysate was the most effective inhibitor of lipid peroxidation. Each sample exhibited a different polypeptide profile. Different percentages of carbohydrates were identified in whey filtrate and in all second fractions, where galactose was the major component.

Antimicrobially active hederagenin glycosides from Cephalaria elmaliensis.

A phytochemical investigation of the aerial parts of Cephalaria elmaliensis resulted in the isolation of ten hederagenin-type triterpene saponins (1-10) including three new ones, elmalienoside A (1), elmalienoside B (2), elmalienoside C (3), and two known flavonoid glycosides (11-12). Their structures were identified by extensive spectroscopic techniques (1D and 2D NMR, HR ESIMS) and chemical evidence. The antimicrobial activity of the extracts and the pure compounds was evaluated by the MIC method. According to the results, all pure compounds including the new ones were found to be very active against both gram-positive and gram-negative bacteria.

A novel glycoside from Acanthus hirsutus (Acanthaceae).

A novel glycoside, hirsutusoide (1), characterized as 2-(o-hydroxyphenyl)-2-hydroxyethenyl-O-beta-glucopyranoside, was isolated from the endemic Acanthus hirsutus Boiss. In addition to compound 1, three known glycosides, luteolin-7-O-beta-D-glucuronide (2), beta-sitosterol-3-O-beta-D-glucopyranoside (3) and (2R)-2-O-beta-D-glucopyranosyl-2H-1,4-benzoxazin-3(4H)-one (4), were also isolated. Compound 2 was the first report from this genus. Antimicrobial and antioxidant activity of the extracts and the novel compound were investigated by determining MIC (microg/mL) and IC50 (microg/mL) values, respectively.

Antimicrobial triterpenoid glycosides from Cephalaria scoparia.

Five new oleanane-type triterpenoid glycosides (1-5), named scoposides A-E, along with one known bisdesmosidic triterpene glycoside, were isolated from the aerial parts of Cephalaria scoparia. Three prosapogenins (2a-4a) were also obtained after alkaline hydrolysis of the bisdesmosidic compounds 2-4. The structures of compounds 1-5 were determined by spectroscopic (1D and 2D NMR, HRESIMS) and chemical methods. The antimicrobial effects of compounds 1-5 were evaluated against a small panel of Gram-positive and Gram-negative bacteria.

Triterpene and flavone glycosides from Anchusa undulata subsp. hybrida.

A novel triterpene glycoside, undulatoside (1), which is characterised as 3-O-(beta-D-glucopyranosyl)-29-O-(beta-D-glucopyranosyl)-2alpha,23-dihydroxyolean-12-en-28-oic acid, was isolated from Anchusa undulata subsp. hybrida and its structure was deduced by spectral data. In addition to undulatoside, seven known triterpene glycoside and a flavonoid glycoside were also isolated for the first time from this plant. The structures have been identified by spectroscopic and chemical methods. The antimicrobial activities of all extracts of the plant were investigated by disc diffusion method.

Flavonoid glycosides and methylinositol from Ebenus haussknechtii.

Two flavonoid glycosides (compounds 1 and 3) of which one is reported for the first time and a methylinositol (compound 2) were isolated from the aerial parts of Ebenus haussknechtii (Leguminosae). The structures were established as quercetin-7-O-[alpha-L-rhamnopyranosyl(1 --> 6)-beta-D-galactopyranoside] (1), morin-3-O-[4-[5-(4-hydroxyphenyl)pentanoyl]-alpha-L-rhamnopyranosyl(1 --> 6)-beta-D-galactopyranosyl]-7-4'-di-O-methyleter (3), and methylinositol (2) on the basis of chemical and spectroscopic means. The antimicrobial activities of the extracts have also been examined.