Simultaneous determination of 7 components in Dipsacus asper by HPLC / 中国药房

OBJECTIVE To establis h the method for the simultaneous determination of six iridoids （loganic acid ，loganin， sweroside，dipsanoside B ，dipsanoside A ，sylvestroside Ⅰ）and one triterpene saponin （asperosaponin Ⅵ）in Dipsacus asper . METHODS High performance liquid chromatography （HPLC） method was adopted. The determination was performed on Symmetry® C18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelengths were set at 212 nm（asperosaponin Ⅵ）and 237 nm（dipsanoside B ，dipsanoside A ， sweroside，loganic acid ，sylvestroside Ⅰ，loganin）. The column temperature was set at 30 ℃，and sample size was 20 μL. RESULTS The linear range of loganic acid ， loganin， sweroside， sylvestroside Ⅰ ， dipsanoside B ， dipsanoside A and asperosaponin Ⅵ were 399.24-931.56，50.30-150.90，48.24-168.84，27.00-70.20，12.93-38.80，40.64-121.92，42.08-147.28 µg/mL （all r＞0.999 0）. RSDs of precision ，reproducibility and stability tests （24 h）were all less than 2％. Average recoveries were 104.43％（RSD＝0.63％，n＝6），101.74％（RSD＝1.11％，n＝6），100.76％（RSD＝1.06％，n＝6），98.00％（RSD＝1.58％，n＝6）， 99.03％（RSD＝2.31％，n＝6），102.93％（RSD＝2.26％，n＝6），102.31％（RSD＝1.00％，n＝6），respectively，The contents were 142.5-280.6，5.5-49.0，28.0-112.9，7.2-35.8，4.4-16.9，17.2-79.3，0.8-54.5 mg/g，respectively. CONCLUSIONS Established method is accurate and reliable ，and can be used for the content determination of 7 components in D. asper .

Content changes of triterpene saponins in crude and sweated Dipsacus asper:an iTRAQ-based analysis / 中国中药杂志

The present study aimed to explore the mechanism of the sweating of Dipsacus asper on content changes of triterpene sa-ponins by detecting the total triterpene saponins and the index component asperosaponin Ⅵ in the crude and sweated D. asper, and analyzing the differentially expressed proteins by isobaric tags for relative and absolute quantification(iTRAQ) combined with LC-MS/MS. After sweating, the content of total triterpene saponins decreased manifestly, while that of asperosaponin Ⅵ increased significantly. As revealed by the iTRAQ-LC-MS/MS analysis, 140 proteins with significant differential expression were figured out, with 50 up-regulated and 90 down-regulated. GO analysis indicated a variety of hydrolases, oxido-reductases, and transferases in the differential proteins. The results of activity test on two differentially expressed oxido-reductases were consistent with those of the iTRAQ-LC-MS/MS analysis. As demonstrated by the analysis of enzymes related to the triterpene saponin biosynthesis pathway, two enzymes(from CYP450 and UGT families, respectively, which are involved in the structural modification of triterpene saponins) were significantly down-regulated after sweating. The findings suggested that sweating of D. asper presumedly regulated triterpene saponins by affecting the expression of downstream CYP450 s and UGTs in the biosynthesis pathway of triterpene saponins of D. asper.

Study on chemical constituents in acid hydrolysates of Panax notoginseng saponins / 中草药

Objective: To study the chemical constituents in acid hydrolysates of Panax notoginseng saponins (PNS). Methods: These compounds were separated and purified by column chromatography, and their structures were elucidated based on spectroscopic analyses (HR-ESI-MS, ESI-MS, 1H-NMR, 13C-NMR, HSQC and HMBC). Results: Eighteen compounds were obtained from the acid hydrolysates of PNS and characterized as dammar-25-ene-24-hydroperoxyl-3β,6α,12β,20S-tetraol (1), 6α,12β,20S-trihydroxy- dammarane-24-ene-3-O-β-D-glucopyranosyl-(1→2)-β-D-glucopyranoside (2), 6α,12β,20R-trihydroxy-dammarane-24-ene-3-O-β-D- glucopyranosyl-(1→2)-β-D-glucopyranoside (3), vina-ginsenoside-R8 (4), 24(S)-pseudo-ginsenoside-GQ (5), ginsenoside Rg5 (6), 20 (R)-ginsenoside Rg3 (7), 20(R)-ginsenoside Rk2 (8), 3β,12β-dihydroxy-dammar-(E)-20(22),24-diene-6-O-β-D-xylopyranosyl- (1→2)-β-D-glucopyranoside (9), 20(S)-ginsenoside Rg2 (10), ginsenoside SL1 (11), 20(R)-ginsenoside Rh1 (12), 20(22) E-ginsenoside Rh4 (13), 25-hydroxy-20(R) ginsenoside-Rh1 (14),3β,6α,12β,20(S)-20,25-epoxy-3,12-dihydroxy-dammarane-6-O-β-D-glucopyranoside (15), 20(R)-protopanaxadiol (16), 20(R)-protopanaxatriol (17), and 20(S)-protopanaxatriol (18). Conclusion: Compound 1 is a new triterpen saponin, and compounds 2-5 are isolated from P. notoginseng and acid dydrolysates of PNS for the first time.

Triterpene saponins in Panax japonicus and their ~(13)C-NMR spectroscopic characteristics / 中国中药杂志

Panax japonicus( PJ) is a valuable medicinal plant belonging to the genus Panax of Araliaceae,the recumbent rhizome of which is widely used in clinic therapy,healthcare products and as cosmetic additives with functions of dissipating stasis,reducing swelling,stanching bleeding,and reinforcing deficiency,etc. PJ contains abundant levels of oleanane-and dammarane-type triterpene saponins,which are considered as the material basis for exerting pharmacodynamic action. Based on the previous researches,more than110 triterpene saponins have been reported from PJ. These triterpene saponins were summarized in this review,and could be classified into dammarenediol Ⅱ,protopanaxadiol,protopanaxatiol,ocotillol,oleanolic acid,ursolic acid and miscellaneous subtypes,according to their molecular skeletons in biosynthesis processes. Further more,the structural features of these triterpene saponins in the seven different subtypes,together with their~(13)C-NMR spectroscopic characteristics were described,hoping to provide available information for chemical diversity research of PJ.

Triterpene saponins from Aralia taibaiensis / 中草药

Objective: To study the triterpene saponins of Aralia taibaiensis. Methods The chemical components were separated and purified by Silica gel, Sephadex LH-20, and sp-HPLC; Their structures were elucidated by 1D/2D NMR, MS, and IR spectral data. Results Eight triterpene saponins were isolated and elucidated as taibaienoside IX (1), chikusetsusaponin 1 (2), chikusetsusaponin Iva (3), lucyoside H (4), narcissiflorine (5), araloside A (6), araliasaponin II (7), and 3-O-[β-D-glucopyranosyl-(1→2)-α-L- arabinopyranosyl]-oleanolic acid-28-O-[β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl] ester (8). Conclusion Compound 1 is a new oleanolic type triterpene saponin which named as taibaienoside IX, while compounds 4 and 8 are isolated from this plant for the first time.

Study of heterologous efficient synthesis of β-amyrin and high-density fermentation / 中国中药杂志

In this study, the synthetic pathway of β-amyrin was constructed in the pre-constructed Saccharomyces cerevisiae chassis strain Y0 by introducing β-amyrin synthase from Glycyrrhiza uralensis, resulting strain Y1-C20-6, which successfully produced β-amyrin up to 5.97 mg·L~(-1). Then, the mevalonate pyrophosphate decarboxylase gene(ERG19), mevalonate kinase gene(ERG12), 3-hydroxy-3-methylglutaryl-CoA synthase gene(ERG13), phosphomevalonate kinase gene(ERG8) and IPP isomerase gene(IDI1)were overexpressed to promoted the metabolic fluxto the direction of β-amyrin synthesis for further improving β-amyrin production, resulting the strain Y2-C2-4 which produced β-amyrin of 10.3 mg·L~(-1)under the shake flask fermentation condition. This is 100% higher than that of strain Y1-C20-6, illustrating the positive effect of the metabolic engineering strategy applied in this study. The titer of β-amyrin was further improved up to 157.4 mg·L~(-1) in the fed-batch fermentation, which was almost 26 fold of that produced by strain Y1-C20-6. This study not only laid the foundation for the biosynthesis of β-amyrin but also provided a favorable chassis strain for elucidation of cytochrome oxidases and glycosyltransferases of β-amyrin-based triterpenoids.

Triterpenoids isolated from flower buds of Lonicera macranthoides / 中草药

Objective To study the chemical constituents from the flower buds of Lonicera macranthoides and their antitumor activities. Methods The constituents were separated by chromatography of silica gel, ODS, Sephadex LH20, and semi-pre HPLC. Their structures were elucidated by spectral means. The in vitro cytotoxic activities of the isolated compounds were studied by MTT method. Results Seven compounds were isolated and identified as 3-O-β-D-glucopyranosyl-(1→4)-α-L-arabinopyranosyl- hederagenin 28-O-β-D-glucopyranosyl ester (1), 3-O-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl-oleanolic acid 28-O-α- L-rhamnopyranosyl-(1→4)-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (2), 3-O-α-L-rhamnopyranosyl-(1→2)-α-L- arabinopyranosyl-hederagenin 28-O-β-D-glucopyranosyl ester (3), 3-O-α-L-rhamnnopyranosyl-(1→2)-α-L-arabinopyranosyl- hederagenin 28-O-α-L-rhamnopyransyl-(1→4)-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (4), 3-O-α-L-arabinopyranosyl- hederagenin 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (5), 3-O-β-D-glucopyra- nosyl-(1→4)-α-L-arabinopyranosyl-hederagenin 28-O-α-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside ester (6), and 3-O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→2)-α-L-arabinopyranosyl-hederagenin 28-O-α-L-rhamno- pyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosyl ester (7). Conclusion Compound 1 is a new compound named macranthoidin C, and compounds 2-7 are isolated from L. macranthoides for the first time. Compounds 1, 4, and 5 show cytotoxicities against HeLa cells with IC50 of 54.3, 43.9 and 61.2 μmol/L, respectively.

Novel triterpene saponins isolated from Clematis mandshurica and their inhibitory activities on NO production / 中国天然药物

Four new triterpene saponins, mandshunosides F-I (1-4), together with five known compounds (5-9), were isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of spectroscopic evidences and hydrolysis products. Bisdesmosidic saponin (3-9) showed modest suppression of NO production with the inhibition ratios in the range of 51.3%- 64.6% at 50 μmol·L, whereas monodesmosidic saponins with a free carboxyl group at C-28 (1 and 2) showed potent inhibitory activities with IC values being 12.7 and 8.3 μmol·L, respectively.

Novel triterpene saponins isolated from Clematis mandshurica and their inhibitory activities on NO production / 中国天然药物

Four new triterpene saponins, mandshunosides F-I (1-4), together with five known compounds (5-9), were isolated from the roots and rhizomes of Clematis mandshurica. Their structures were elucidated on the basis of spectroscopic evidences and hydrolysis products. Bisdesmosidic saponin (3-9) showed modest suppression of NO production with the inhibition ratios in the range of 51.3%- 64.6% at 50 μmol·L, whereas monodesmosidic saponins with a free carboxyl group at C-28 (1 and 2) showed potent inhibitory activities with IC values being 12.7 and 8.3 μmol·L, respectively.

Rapid simultaneous determination of nine triterpene saponins constituents of Caulophyllum robustum by UPLC-FT/MS / 中草药

Objective To establish a rapid simultaneously determination of nine triterpene saponins of Cauorlhyllum robustum by UPLC-FT/MS. Methods UPLC-MS/MS assay in positive ion mode was performed on a ACE Excel 3 C18 (100 mm × 2.1 mm, 3.0 μm) with the mobile phase consisting of 0.1% formic acid aqueous (A) and 0.1% formic acid acetonitrile (B) in gradient elution at a flow rate of 0.40 mL/min and the column temperature was set at 35 ℃. Results Satisfactory linearity was achieved with wide linear range and fine linear relationship (r > 0.996 3), the recoveries were ranged from 98.49% to 107.03% with the RSD ranging from 1.00% to 3.33%. Conclusion It is the first report about simultaneous analysis of nine triterpene saponins of Cauorlhyllum robustum by UPLC-FT/MS method, which affords highly sensitive, resolution, simple, accurate, and speedy efficient method for quality control of Cauorlhyllum robustum.

Leishmanicidal and trypanocidal activity of Sapindus saponaria / Actividad leishmanicida y tripanocida de Sapindus saponaria

Leishmaniasis and trypanosomiasis are parasitic diseases with a high infection rate, being a serious public health issue in the new world. Unfortunately, there are few available commercial drugs, poorly efficient and with increasing parasite resistance. Under these condi- tions, there is a need for new molecules to develop new and better drugs. One approach to carry out this search is using traditional medicine as information source to obtain new molecules or extracts to control these parasite diseases. Sapindus saponaria (Sapindaceae) fruit resin is used in Colombia to treat ulcers caused by Leishmaniasis. In a bioguided study, we have analyzed the in vitro effect of fruit resin, chroma- tographical fractions from fruit resin and also pure compounds against Leishmania species (L. panamensis, L. braziliensis, L. amazonensis and L. donovani) and Trypanosoma cruzi. The in vivo antileishmanial effect was established under the hamster model for cutaneous leish- maniasis by L. panamensis; refined extract of S. saponaria and pure saponins displayed high in vitro and in vivo activity as leishmanicides. In addition, extracts caused low viability on T. cruzi amastigotes. The use of the crude extract can be a good alternative against cutaneous leishmaniasis, due to its activity, reduced hemolytic effect, and easy production procedures.

La Leishmaniasis y la tripanosomiasis son enfermedades parasitarias con una alta incidencia, siendo un serio asunto de salud pública en el nuevo mundo. Desafortunadamente, hay pocas drogas comerciales disponibles, con pobre eficiencia y con una creciente resis- tencia parasitaria. Bajo esas condiciones, se necesitan nuevas moléculas para desarrollar nuevas y mejores drogas. Una aproximación para llevar a cabo esa búsqueda es usar la medicina tradicional como fuente de información para obtener nuevas moléculas o extractos para con- trolar esas enfermedades parasitarias. La resina de Sapindus saponaria (Sapindaceae) se usa en Colombia para tratar úlceras causadas por la Leishmaniasis. En un estudio bioguiado, se analizó el efecto in vitro de varios extractos de la resina, sus fracciones cromatográficas y algu- nos compuestos puros, contra varias especies de Leishmania (L. panamensis, L. braziliensis, L. amazonensis y L. donovani) panamensis y Trypanosoma cruzi. El efecto lesihmanicida in vivo fue establecido usando el modelo en hamster de leishmaniasis cutánea producida por L. panamensis; los extractos refinados de S. saponaria y las saponinas puras mostraron alta actividad in vitro e in vivo como leishmanicidas. Además, los extractos causaron una baja viabilidad en amastigotes de T. cruzi. El uso de extractos refinados en vez de saponinas puras podría ser una buena alternativa contra leishmaniasis cutánea debido a su actividad, poco efecto hemolítico y procedimientos de producción mucho más fáciles.