Purification of triterpenoid saponins and 25R/25S-inokosterone from Achyranthes bidentata Bl. by high-speed countercurrent chromatography coupled silver nitrate coordination.

An effective method by high-speed countercurrent chromatography coordinated with silver nitrate for the preparative separation of sterones and triterpenoid saponins from Achyranthes bidentata Blume was developed. Methyl tert-butyl ether/n-butanol/acetonitrile/water (4:2:3:8, v/v/v/v) was selected for 20-hydroxyecdysone (compound 1), chikusetsusaponin IVa methyl ester (compound 4), 2'-glycan-11-keto-pigmented saponin V (compound 5), as well as a pair of isomers of 25S-inokosterone (compound 2) and 25R-inokosterone (compound 3), which were further purified by silver nitrate coordinated high-speed countercurrent chromatography. What is more, dichloromethane/methanol/isopropanol/water (6:6:1:4, v/v/v/v) was applied for calenduloside E (compound 6), 3ß-[(O-ß-d-glucuronopyranosyl)-oxy]-oleana-11,13-dien-28-oic acid (compound 7), zingibroside R1 (compound 8) and chikusetsusaponin IVa (compound 9). Adding Ag+ to the solvent system resulted in unique selectivity for 25R/25S isomers of inokosterone, which increased the complexing capability and stability of Ag+ coordinated 25S-inokosterone, as well as the α value between them. These results were further confirmed by the computational calculation of geometry optimization and frontier molecular orbitals assay. Comprehensive mass spectrometry and nuclear magnetic resonance analysis demonstrated the structures of the obtained compounds.

[A new sesquiterpenoid from Lindera aggregata].

The chemical composition of the aqueous part of the extract from Lindera aggregata was studied, which was separated and purified by the macroporous resin column chromatography, MCI medium pressure column chromatography, semi-preparative high-performance liquid phase and other methods. The structures of the compounds were identified according to physical and chemical properties and spectroscopic data. Thirteen compounds were isolated and identified from the aqueous extracts, which were identified as(1S,3R,5R,6R,8S,10S)-epi-lindenanolide H(1), tachioside(2), lindenanolide H(3), leonuriside A(4), 3,4-dihydroxyphenyl ethyl ß-D-glucopyranoside(5), 3,4,5-trimethoxyphenol-1-O-6-α-L-rhamnose-(1→6)-O-ß-D-glucoside(6), 5-hydroxymethylfurfural(7),(+)-lyoniresin-4-yl-ß-D-glucopyranoside(8), lyoniside(9), norboldine(10), norisopordine(11), boldine(12), reticuline(13). Among them, compound 1 was a new one, and compounds 2, 5, 6, 8, 9 were obtained from L. aggregata for the first time. The inflammatory model was induced by lipopolysaccharide(LPS) in the RAW264.7 cells. The results showed that compounds 1, 8, 10 and 12 had significant anti-inflammatory activity.

Nomogram for predicting early hypophosphatemia in term infants.

BACKGROUND: Physiological processes rely on phosphate, which is an essential component of adenosine triphosphate (ATP). Hypophosphatasia can affect nearly every organ system in the body. It is crucial to monitor newborns with risk factors for hypophosphatemia and provide them with the proper supplements. We aimed to evaluate the risk factors and develop a nomogram for early hypophosphatemia in term infants. METHODS: We conducted a retrospective study involving 416 term infants measured serum phosphorus within three days of birth. The study included 82 term infants with hypophosphatemia (HP group) and 334 term infants without hypophosphatemia (NHP group). We collected data on the characteristics of mothers, newborn babies, and childbirth. Furthermore, univariate and multivariate logistic regression analyses were performed to identify independent risk factors for hypophosphatemia in term infants, and a nomogram was developed and validated based on the final independent risk factors. RESULTS: According to our analysis, the multivariate logistic regression analysis showed that male, maternal diabetes, cesarean delivery, lower serum magnesium, and lower birth weight were independent risk factors for early hypophosphatemia in term infants. In addition, the C-index of the developed nomogram was 0.732 (95% CI = 0.668-0.796). Moreover, the calibration curve indicated good consistency between the hypophosphatemia diagnosis and the predicted probability, and a decision curve analysis (DCA) confirmed the clinical utility of the nomogram. CONCLUSIONS: The analysis revealed that we successfully developed and validated a nomogram for predicting early hypophosphatemia in term infants.

Efficient discovery of active isolates from Dioscorea spongiosa by the combination of bioassay-guided macroporous resin column chromatography and high-speed counter-current chromatography.

In the present study, twelve compounds from Dioscorea spongiosa were successfully purified by an efficient technique combined bioassay-guided fractionation macroporous resin column chromatography (MRCC) pretreatment and high-speed counter-current chromatography (HSCCC) separation for the first time. Then, D101 MRCC was used to fractionate the crude extract into five parts, which further applied the bioassay-guided fractionation strategy to screen the active fractions of 2 and 4. As for the separation, 200 mg Fr.2 was purified by HSCCC using EtOAc/n-BuOH/H2 O (2:2:3, v/v), leading to annulatomarin (1), dioscoresides C (2), diosniponol C (3), methyl protodioscin (4), pseudoprotodioscin (5), protogracillin (6), as well as 200 mg Fr.4 yielding montroumarin (7), dioscorone A (8), diosniponol D (9), protodioscin (10), gracillin (11), and dioscin (12) using CH2 Cl2 /MeOH/H2 O (3:3:2, v/v) with the purities over 95.0%. Finally, the isolates were assayed for their anti-inflammatory, urico-lowering, and anti-diabetic activities in vitro, which indicated that the steroidal saponins of 5, 6, and 11 showed all these three activities.

Correlations between the structure and anti-diabetic activity of novel polysaccharides from raw and "Nine Steaming Nine Sun-Drying" Polygonti rhizome.

"Nine Steaming Nine Sun-Drying" Polygonti rhizome has been used as valuable tonic health-care products for thousands of years. This research aimed to determine the correlations between the structure and anti-diabetic activities of three novel polysaccharides isolated from the raw and "Nine Steaming Nine Sun-Drying" Polygonti rhizome, with PRP-R from the raw ones and PRP-9Z and PRP-9A from the steamed ones. Structures of the isolated polysaccharides were determined by IR and NMR spectra, as well as monosaccharide composition and methylation analysis. In vitro assays indicated that PRP-9Z could improve the glucose consumptions more effectively than PRP-R and PRP-9A via Akt/GSK3ß insulin signal pathway by western blotting analysis. In vivo assays indicated PRP-9Z could improve the glucose tolerance in the BKS-db mice. Histopathological assay demonstrated that PRP-9Z effectively reduced the damage of the kidney and liver. The above results indicated that PRP-9Z from "Nine Steaming Nine Sun-Drying" Polygonti rhizome showed significant anti-diabetic properties, which indicated that PRP-9Z with higher content of →1)-ß-Frup-(2→ was more active than PRP-R with higher →1,6)-ß-Fruf-(2→ and PRP-9A with higher →4)-ß-Galp(1→.

Secondary metabolites from Scindapsus officinalis (Roxb.) Schott. with in vitro antidiabetic activities.

One new phenolic cyclobutantetraol ester united chromone glycoside (1), one new amide (2), and three new phenyl ethanol glycosides (3-5) were obtained from the water extract of Scindapsus officinalis (Roxb.) Schott, in which compound 1 was the first reported structure incorporating the phenolic cyclobutantetraol ester and chromone via the glucose phenolic metabolites in nature. Structures of the isolated compounds, including absolute configurations, were elucidated according to the analysis of HRESIMS, NMR, ECD and BLYP/6-31G* geometry optimization calculations of 13C NMR data. All isolates (1-5) were evaluated for the antidiabetic activity by the insulin resistance (IR) model and anti-inflammatory activity against NO production in vitro. Compounds 1-3 showed strong antidiabetic activities, greatly promoting the glucose consumption in the insulin resistance HepG2 cells compared with the model group, however, 1-5 showed weak anti-inflammatory activities.

Separation and purification of alkaloids and phenolic acids from Phellodendron chinense by pH-zone refining and online-storage inner-recycling counter-current chromatography.

In this work, eight compounds from Phellodendron chinense were separated and purified by pH-zone refining counter-current chromatography and traditional counter-current chromatography coupled with online-storage inner-recycling counter-current chromatography (IRCCC). The pH-zone-refining mode was adopted for separating 2.0 g of crude extract with the solvent system of chloroform-methanol-water (4:3:3, v/v), in which 10 mM hydrochloric acid and 10 mM triethylamine were added in the stationary and mobile phases, respectively. Meanwhile, traditional counter-current chromatography coupled with online-storage IRCCC separation was performed by the solvent system of n-hexane-ethyl acetate-methanol-water (5:5:2:8, v/v). Finally, eight compounds, including six alkaloids as 6-methylpiperidin-2-one(1), isoplatydesmine(4), berlambine(5), epiberberine(6), palmatine(7), berberine(8) and two phenolic acids as ferulic acid(2), isoferulic acid(3), were successfully obtained using these three different CCC modes with the purities over 95.0%.

Cytotoxic constituents from the leaves and stems of Panax quinquefolius.

Four new isolates including one new butanediamide glycoside (1), one new flavonoid glycoside (2) and two new flavonone glycosides (3, 4) were identified from the leaves and stems of Panax quinquefolius, among which 1 possessed the firstly reported N,N'-(5-hydroxy-1,3-phenylene) butanediamide skeleton with an unique 6/9 ranged dual-ring structure. The structures were elucidated by the NMR data, ECD analysis and chemical acid hydrolysis. All the compounds (1-4) were tested for their cytotoxicity against two human cancer cell lines of HepG2, A549 and HCT116 by the MTT method. Outstandingly, compound 1 exhibited targeted inhibitory proliferation of HCT116 cell with IC50 value of 12.1 µM, whereas compounds 3 and 4 exhibited targeted inhibitory proliferation of HepG2 cell with IC50 values of 15.3 and 17.3 µM, as well as no obvious cytotoxicity of compounds 1-4 against A549.

Efficient separation of anti-inflammatory isolates from Polygonti rhizome by three different modes of high-speed counter-current chromatography.

Successful isolation of 15 compounds from Polygonti rhizome was obtained by an efficient technique combined with macroporous resin column chromatography pretreatment and three different modes of high-speed counter-current chromatography for the first time. For the pretreatment, AB-8 resin was applied to remove the polysaccharides and enrich four different parts (samples I, II, III, and IV) by polarities. For the separation, sample I was separated by pH-zone-refining counter-current chromatography and seven cycle recycling mode high-speed counter-current chromatography, yielding four alkaloids (1--4); samples II-IV were further separated by the conventional high-speed counter-current chromatography, yielding seven flavonoids (5-10, 12), one steroid saponin (11), and three terpenoids (13-15). Finally, the isolates were assayed for their anti-inflammatory activities against nitric oxide production with compounds 5, 9-10, 13 showing significant anti-inflammatory activities, IC50 values which were 13.0, 16.2, 17.1, and 14.7 µM, respectively, while others showing moderate and weak anti-inflammatory activities, respectively.

Bioactive constituents from the leaves of Lonicera japonica.

Six new isolates including three new alkaloids (1-3), one new secoiridoid glycoside (4) and one new 11-delactonization-secoiridoid glycoside (5), and one new organic acid (6) were identified from the leaves of Lonicera japonica, among which 1 and 2 assigned as a pair of configurational isomers possessed two oxazolidin-2-one fragments connected through NN bond. The structures were elucidated by the NMR data and ICD analysis. All the compounds (1-6) were tested for their antioxidant and hepatoprotective activities using cell models of ABAP-induced HepG2 cell and APAP-induced HepG2 cell by the MTT method. Outstandingly, compound 2 exhibited remarkable antioxidative activity with inhibitory rate of 117.2%, and compounds 2, 4, 6 exhibited significant effects with inhibitory rates of 68.1%, 69.3%, 69.2%, respectively.

Weed Suppression and Molecular Mechanisms of Isochlorogenic Acid A Isolated from Artemisia argyi Extract via an Activity-Guided Method.

Allelopathy is considered an environmentally friendly and resource-conserving approach to weed control because allelochemicals degrade easily and cause less pollution than traditional chemical herbicides. In this study, the allelopathic active constituents of Artemisia argyi were elucidated by activity-guided isolation and ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS). First, a crude extract prepared in water was fractionated using macroporous resin D101 to obtain three fractions (Fr.A-C). Combined with the allelopathic activity assay on Setaria viridis and Portulaca oleracea, Fr.C was determined to be the most active fraction. We identified 14 compounds in the active fraction (Fr.C) using UPLC-QTOF-MS, including 13 phenolic compounds. Accordingly, phenolic components have been suggested as the main allelochemicals in A. argyi. Thereafter, Fr.C was further isolated by octadecylsilyl (ODS) chromatography to obtain eight subfractions (Fr.C-1-Fr.C-8). Finally, isochlorogenic acid A (ICGAA) was purified from Fr.C-3 by semipreparative liquid chromatography, which was detected in the growth environment of A. argyi. Furthermore, we evaluated the allelopathic effects of ICGAA on six weeds from different families and genera for the first time. The results showed that ICGAA is a novel allelochemical with broad herbicidal activity. In addition, we analyzed the inhibitory effect and molecular mechanism of ICGAA on the growth of S. viridis seedlings. Optical microscopy and transmission electron microscopy (TEM) revealed the degradation of membrane structures and organelles after ICGAA treatment. Transcriptome and real-time polymerase chain reaction (RT-qPCR) analysis showed that ICGAA inhibited the growth of weeds mainly by inhibiting the diterpenoid biosynthesis pathway (especially gibberellins, GAs). The decrease of gibberellin (GA) contents after ICGAA treatment also confirmed these results. In brief, this study provides new material sources and theoretical support for developing biological herbicides for agroecosystems.

Anti-inflammatory and hepatoprotective cembranoid alcohols from the Gum Resin of Boswellia carterii.

Five undescribed cembranoid alcohols, boscartinols A-E (1-5) were discovered from the gum resin of Boswellia carterii. Their structures were elucidated by analyzing the spectroscopic data. Notably, all these five compounds preserved an isopropyl type cembranoid skeleton, featured the same groups of one epoxy ring at C1-C12 and one hydroxy group at C-11, as well as two double bonds migrating from C3 to C9 and one hydroxy group from C3 to C8 within the cembranoid structure. These cembranoids were evaluated for the hepatoprotective and anti-inflammatory activities using two cell models of APAP-induced HepG2 and LPS-induced RAW 264.7. For hepatoprotective activity, compounds 1 and 5 showed remarkable hepatoprotective activity (inhibition rate of 51.6% and 39.8%, respectively) at 10 µM, with the other three compounds of 2-4 showing less potently hepatoprotective. For anti-inflammatory activity, compounds 2-4 showed significant inhibitory effects on NO produced by LPS-induced RAW 264.7 cell (IC50 values of 13.40 µM, 7.08 µM and 14.26 µM), with the other two compounds of 1 and 5 showing less potently anti-inflammatory activities.

Application of choline chloride deep eutectic solvents and high-speed counter-current chromatography to the extraction and purification of flavonoids from the thorns of Gleditsia sinensis Lam.

INTRODUCTION: Flavonoids are the most important and effective constituents in the thorns of Gleditsia sinensis Lam., which have been known to show antimicrobial, antiviral, anticancer, and anticoagulant activities. However, efficient extraction and separation methods for these flavonoids are not currently established. OBJECTIVE: To develop an efficient method for efficient extraction and rapid separation of flavonoids from the thorns of G. sinensis using choline chloride deep eutectic solvents (DESs) and high-speed counter-current chromatography (HSCCC). METHODOLOGY: As for extraction, DES composed of choline chloride and 1,4-butanediol at 1:4 mole ratio, at an extraction temperature of 55°C, 20% of water content, 1:30 mg/mL for solid-liquid ratio, and 45 min for extraction time were selected as the optimised extraction method for flavonoids from the thorns of G. sinensis. As for separation, dichloromethane-methanol-n-butanol-water (4:3:0.5:2, v/v) was applied to develop a successful strategy for purification of the flavonoids by HSCCC. RESULTS: Totally, five flavonoids, including padmatin (1, 3.7 mg), isovitexin (2, 2.5 mg), 3',5,5',7-tetrahydroxyflavanonol (3, 11.2 mg), 7,4'-dihydroxy-5,3'-dimethoxyflavanonol (4, 4.1 mg), and quercetin (5, 3.8 mg), were successfully obtained from 250 mg of the extracted flavonoids by HSCCC. CONCLUSION: Results demonstrated that the combination of DES and HSCCC is a powerful technique for the extraction, and isolation of flavonoids from the thorns of G. sinensis compared with conventional organic solvent extraction and column chromatography, which have been proven to provide higher extraction efficiency for flavonoids and rapidly obtain the quality control markers of flavonoids from the investigated plant.

Bioactive cembrane diterpenoids from the gum resin of Boswellia carterii.

Eight new cembrane-type diterpenoids, boscartins AP-AW (1-8) were obtained from the gum resin of Boswellia carterii. Among which, six ones (2-7) were isomers, with one hydroxy group and two double bonds migrating along the carbocycle. The structures were elucidated by spectroscopic examination. All isolates were evaluated for anti-inflammatory activity and hepatoprotective activity by cell models of LPS-induced RAW 264.7 mouse peritoneal macrophages and APAP-induced HepG2 cells, respectively. As for anti-inflammatory activity assay, compound 1 exhibited potent activity against NO production (IC50 of 13.1 µM), with the other ones exhibiting weak anti-inflammatory activity (IC50 > 50 µM). As for hepatoprotective activity assay, compound 1 exhibited more significant activity (inhibition rate of 30.7%) than that of the positive control (bicyclol, inhibition rate of 27.2%), and compounds 4, and 6 showed nearly the same activities as the control (inhibition rates of 26.7% and 25.9%, respectively), with the other ones exhibiting weak hepatoprotective activity.

Flavonoid epimers from custard apple leaves, a rapid screening and separation by HSCCC and their antioxidant and hypoglycaemic activities evaluation.

Leaves of custard apple are widely used in many places as a popular dietary supplement for the treatment of diabetes. Flavonoids are known to have anti-diabetic activity. In this study, the main flavonoid epimers were separated. The crude extract was first screened by HPLC-DAD before and after incubation with DPPH method to evaluate the antioxidants. An efficient extraction method was employed to remove non-flavonoid components. Subsequently, five main flavonoids with two pairs of epimers including quercetin-3-O-robinobioside, rutin, quercetin-3-O-ß-D-glucoside, kaempferol-3-O-robinobioside, and kaempferol-3-O-rutinoside were successfully separated by high-speed counter-current chromatography with ethyl acetate/n-butanol/water (4:1:5, v/v) coupled with online-storage inner-recycling mode. The structures of the separated compounds were identified by spectral techniques. The purity of the separated flavonoid glycosides was over 98%, as determined by HPLC. The separated pure constituents were found to possess the antioxidant capacities following DPPH radical scavenging protocol. The compounds (1-3) exhibited better antioxidant activity. Furthermore, the glucose uptake of crude flavonoid extract had better results than the crude ethanol extract. The present study demonstrates that the efficacy of custard apple leaves in lowering glucose level, and antioxidant capacities of separated pure compounds probably appear to be predominantly responsible for hypoglycaemic properties on HepG2 cells.

Comprehensive separation of iridoid glycosides and triterpenoid saponins from Dipsacus asper with salt-containing solvent by high-speed countercurrent chromatography coupled with recycling mode.

The roots of Dipsacus asper Wall as a commonly used traditional Chinese medicine are used for tonifying liver and kidney and strengthening bones and muscles. However, an effective separation strategy for comprehensive and rapid separation of the main active compounds from the roots of D. asper is nonexistent. This investigation provided an effective separation method based on AB-8 macroporous resin column chromatography using different ratios of ethanol in water and two different modes of high-speed countercurrent chromatography with salt-containing solvent system for rapid enrichment and separation from the roots of D. asper. The macroporous resin column chromatography was performed on AB-8 resin using ethanol in water ratios of 10, 30, 40, 50, and 80% as the optimized enrichment conditions for iridoid glycosides and triterpenoid saponins with different polarities. For high-speed countercurrent chromatography separation, the conventional and recycling modes were combined together to develop a strategy for 12 compounds (1-12) from the enriched parts of 30, 40, and 80% ethanol, including six high-polarity iridoid glycosides (1-6) using inorganic salt-containing solvent system and six triterpenoid saponins (7-12). Recycling high-speed countercurrent chromatography separation was successfully applied to separate two isomers (9 and 10) after 11 cycles.

A simple and efficient linear gradient coupled with inner-recycling high-speed counter-current chromatography mode for the preparative separation of flavonoid glycosides from leaves of custard apple.

In general, the simultaneous separation and isolation of compounds with a broad polarity range from natural products is a challenge by ordinary high-speed counter-current chromatography (HSCCC). Indeed, the complex solvent system screening methods limit the broader application of HSCCC. We describe herein a rapid and efficient linear gradient CCC (LGCCC) method that enables the separation of flavonoid glycosides with a broad range of KD values from custard apple leaves. Inner-recycling CCC (IRCCC) mode has been further applied for the separation of compounds with similar KD values. Similarly to binary gradient HPLC, the LGCCC mode is achieved by adjustment of the proportion between ethyl acetate (pump A) and n-butanol (pump B) in an ethyl acetate/n-butanol/water solvent system. Various separation factors have been investigated, including separation mode, rotation speed, flow rate, and sample loading. The IRCCC mode has been used for the secondary separation of two epimers with a simple ethyl acetate/water (1:1, v/v) solvent system. Finally, five main flavonoid glycosides have been successfully separated, namely quercetin-3-O-robinobioside (1, 4.8 mg) and rutin (2, 12.1 mg), quercetin-3-O-ß-d-glucoside (3, 4.2 mg), kaempferol-3-O-robinobioside (4, 9.6 mg), and kaempferol-3-O-rutinoside (5, 24.6 mg). The purities of the separated flavonoid glycosides were over 98%, as determined by HPLC. Our study indicates that a suitable combination of LGCCC and IRCCC modes is an effective strategy for separating flavonoid glycosides from custard apple leaves. The mathematical expression of the LGCCC was deduced to illuminate the separation mechanism. It may also be applied to obtain component fractions for the further screening of active compounds from complex natural products.

Phenolic cyclobutantetraol esters from Scindapsus officinalis (Roxb.) Schott.

Five new caffeoyl cyclobutantetraol esters (1-5) and one hydroxycinnamoyl cyclobutantetraol ester (6), were isolated from Scindapsus officinalis (Roxb.) Schott, which were the first reported phenolic metabolites incorporating a cyclobutantetraol in nature. Structures of the isolated compounds, including absolute configurations, were elucidated by spectroscopic analysis, especially 2D NMR techniques and exciton chirality CD (ECCD) method. All isolates were evaluated for cytotoxic activity toward MCF-7 human breast cancer cell, anti-inflammatory activity against nitric oxide (NO) production, and their antioxidative activity in the 1,1-diphenyl-2-picrylhydrazyl assay in vitro. Compound 1 showed strong antioxidative activity with IC50 value of 59.2 µM, and compounds 1-6 exhibited weak inhibitory effects on NO production, while hardly showing any cytotoxic effects against MCF-7 cell.

Preparative Separation of Diterpene Lactones and Flavones from Andrographis paniculate Using Off-Line Two-Dimensional High-Speed Counter-Current Chromatography.

Seven diterpene lactones, andrographolide (1), isoandrographolide (2), neo-andrographolide (3), 14-deoxy-11,12-didehydroandrographolide (4), 14-deoxyandrographiside (5), 14-deoxy-11,12-didehydroandrographiside (6), 3,14-dideoxyandrographolide (10), and three flavones, andrographidine C (7), andrographidine A (8), 5-hydroxy-7,8-dimethoxyflavanone (9) have been successfully and efficiently isolated from A. paniculata using an off-line two dimensional (2D) high-speed counter-current chromatography (HSCCC) method for the first time. For the first dimension HSCCC separation, petroleum ether-ethyl acetate-methanol-water 3:7:5:5 (v/v) was employed to isolate 14.4 mg of compound 1, 3.1 mg of compound 2, 7.8 mg of compound 3, and 18.0 mg of compound 4 from 200 mg of the A. paniculata extract. For the second dimension HSCCC separation, petroleum ether-ethyl acetate-methanol-water 2:8:1:9 (v/v) and 5:5:6:4 (v/v) were employed to isolate the collected fractions ranged from 55 to 79 min and the flow out fraction, respectively, which led to 5.1 mg of compound 5, 4.4 mg of compound 6, 2.4 mg of compound 7, 3.3 mg of compound 8, 4.0 mg of compound 9, 7.0 mg of compound 10. The structures of these diterpene lactones and flavones were elucidated by extensive spectroscopic methods.

Anti-breast cancer triterpenoid saponins from the thorns of Gleditsia sinensis.

One new triterpenoid saponin (1), as well as six known ones (2-7), were isolated from the ethanol extract of the thorns of Gleditsia sinensis. Their structures were elucidated by extensive spectroscopic analysis in conjunction with chemical evidence. Cytotoxic activity of compounds 1-6 was evaluated against human breast cancer MCF 7 cells in vitro by the MTT method. Our results revealed moderate activities for compounds 1-6 with IC50 values of 18.43, 30.47, 18.46, 10.02, 30.76, and 17.32 µM, respectively. Furthermore, compounds 1, 3, 4, and 6 induced apoptosis in MCF 7 cell, with 1 and 6 causing late apoptosis of MCF 7 cells, while 3 and 4 acting oppositely.