Determination on Heavy Metals Content of Achyranthes bidentata Blume. through Inductively Coupled Plasma Mass Spectrometry (ICP-MS)

[Objective] The inductively coupled plasma mass spectrometry(ICP-MS)was constructed to determine the contents of lead,cadmium,mercury and arsenic in Archyranthes bidentata Blume.[Method]Under the optimum operation condition of ICP-MS,the samples were digested by microwave.The element 114In was taken as an internal standard element to compensate body effect and ICP-MS method was used to determine the contents of lead,cadmium,mercury and arsenic.[Result]For the determined elements,the correlation coefficient(r)of standard curve was over 0.9995 and recovery rate was from 96.7% to 106.4% while RSD was less than 11.2%.The result of determination showed that the heavy metal content in Archyranthes bidentata Blume.beyond standard was serious.[Conclusion]The constructed ICP-MS method with simple operation,rapid response,accuracy and high sensitivity in this experiment could be used for quality control of Chinese medicinal materials by detecting heavy metal contents in different Chinese medicinal materials from original places.

Analysis of the Constituents and Antisenile Function of Achyranthes bidentata Polysaccharides

Achyranthes bidentata polysaccharides (ABPS), water_soluble polysaccharides, isolated from the roots of Achyranthes bidentata Bl. of Amaranthaceae family, was divided into four parts, named as Con.1, Con.2, Con.3 and Con.4, respectively, by chromatography on DEAE_Sepharose fast_flow column and Sephadex G_100 column in order. Con.1 was the constituent of high molecular weight and the other three were all of low molecular weight. Micro_Kjeldahl analysis showed that Con.1 contained 3.95% of nitrogen and neither did the other three parts. The antisenile effects of the four parts of ABPS were studied with Drosophila melanogaster. Results showed that Con.1 has no antisenile effect and all the others could significantly increase the average body weight by 3.85%-5.47% and significantly prolonged the average lifespan by 2.61%- 3.16% of D. melanogaster at the concentration of 2 or 5 mg/g (ABPS/medium).

Inhibiting Effects of Achyranthes Bidentata Polysaccharide and Lycium Barbarum Polysaccharide on Nonenzyme Glycation in D-galactose Induced Mouse Aging Model

Objective To investigate the inhibiting effects and mechanism of achyranthes bidentata polysaccharide (ABP) and lycium barbarum polysaccharide (LBP) on nonenzyme glycation in D-galactose induced mouse aging model. Methods Serum AGE levels were determined by AGE-ELISA, MTT method was used to determine lymphocyte proliferation, IL-2 activity was determined by a bioassay method. Spontaneous motor activity was used to detect mouse's neuromuscular movement, latency of step-through method was used to examine learning and memory abilities of mouse, colormetric assay was used to determine hydroxyproline concentration in mouse skin, pyrogallol autoxidation method was used to determine superoxide dismutase (SOD) activity of erythrocytes. Results Decreased levels of serum AGE, hydroxyproline concentration in mouse skin and spontaneous motor activity in D-galactose mouse aging model were detected after treated with ABP or LBP, while lymphocyte proliferation and IL-2 activity, learning and memory abilities, SOD activity of erythrocytes, were enhanced. Conclusions ABP and LBP could inhibit nonenzyme glycation in D-galactose induced mouse aging model in vivo and ABP has a better inhibiting effect than LBP.

Tissue-based metabolite profiling and qualitative comparison of two species of Achyranthes roots by use of UHPLC-QTOF MS and laser micro-dissection

Achyranthes bidentata and Achyranthes aspera are saponin and steroid rich medicinal plants, used extensively for therapeutic treatments in Traditional Chinese Medicine(TCM) and Ayurveda. A. bidentata is reported to be one of the rare and extensively exploited medicinal plant species that face the issue of being endangered.Finding qualitative substitute with identical phyto-constituents contributing to similar composition and pharmacological benefits will help in reducing the burden of exploitation of the natural habitats of such plants.In the present study, a comparative metabolite analysis of the whole drug and specific tissues isolated by laser micro-dissection(LMD) was carried out for both the selected species, by use of ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UHPLC-QTOF MS). The results of the study indicate that the cortex and the medullary ray tissues are rich in their content of steroidal and saponin constituents such as(25 S)-inokosterone-20,22-acetonide, ginsenoside Ro, bidentatoside II and achyranthoside B.Metabolite profiling of the whole tissues of both the species indicates presence of identical constituents. Thus,it is inferred that A. bidentata and A. aspera can be used as qualitative substitutes for each other.

Achyranthes bidentata polypeptides prevent apoptosis by inhibiting the glutamate current in cultured hippocampal neurons

Glutamate-induced excitotoxicity plays a critical role in the neurological impairment caused by middle cerebral artery occlusion.Achyranthes bidentata polypeptides have been shown to protect against neurological functional damage caused by middle cerebral artery occlusion,but the underlying neuroprotective mechanisms and the relationship to glutamate-induced excitotoxicity remain unclear.Therefore,in the current study,we investigated the protective effects of Achyranthes bidentata polypeptides against glutamate-induced excitotoxicity in cultured hippocampal neurons.Hippocampal neurons were treated with Mg^2+-free extracellular solution containing glutamate(300μM)for 3 hours as a model of glutamate-mediated excitotoxicity(glutamate group).In the normal group,hippocampal neurons were incubated in Mg^2+-free extracellular solution.In the Achyranthes bidentata polypeptide group,hippocampal neurons were incubated in Mg^2+-free extracellular solution containing glutamate(300μM)and Achyranthes bidentata polypeptide at different concentrations.At 24 hours after exposure to the agents,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and Hoechst 33258 staining were used to assess neuronal viability and nuclear m'orphology,respectively.Caspase-3 expression and activity were evaluated using western blot assay and colorimetric enzymatic assay,respectively.At various time points after glutamate treatment,reactive oxygen species in cells were detected by H2 DCF-DA,and mitochondrial membrane potential was detected by rhodamine 123 staining.To examine the effect of Achyranthes bidentata polypeptides on glutamate receptors,electrophysiological recording was used to measure the glutamate-induced inward current in cultured hippocampal neurons.Achyranthes bidentata polypeptide decreased the percentage of apoptotic cells and reduced the changes in caspase-3 expression and activity induced by glutamate.In addition,Achyranthes bidentata polypeptide attenuated the amplitude of the glutamate-induced current.Furthermore,the glutamate-induced increase in intracellular reactive oxygen species and reduction in mitochondrial membrane potential were attenuated by Achyranthes bidentata polypeptide treatment.These findings collectively suggest that Achyranthes bidentata polypeptides exert a neuroprotective effect in cultured hippocampal neurons by suppressing the overactivation of glutamate receptors and inhibiting the caspase-3-dependent mitochondrial apoptotic pathway.All animal studies were approved by the Animal Care and Use Committee,Nantong University,China(approval No.20120216-001)on February 16,2012.

The Achyranthes bidentata polypeptide k fraction enhances neuronal growth in vitro and promotes peripheral nerve regeneration after crush injury in vivo

We have previously shown that Achyranthes bidentata polypeptides （ABPP）, isolated from Achyranthes bidentata Blume （a medicinal herb）, exhibit neurotrophic and neuroprotective effects on the nervous system. To identify the major active component of ABPP, and thus optimize the use of ABPP, we used reverse-phase high performance liquid chromatography to separate ABPP. We obtained 12 fractions, among which the fraction of ABPPk demonstrated the strongest neuroactivity. Immunocytochemistry and western blot analysis showed that ABPPk promoted neurite growth in cultured dorsal root ganglion explant and dorsal root ganglion neurons, which might be associated with activation of Erk1/2. A combination of behavioral tests, electrophysiological assessment, and histomorphometric analysis indicated that ABPPk enhanced nerve regeneration and function restoration in a mouse model of crushed sciatic nerve. All the results suggest that ABPPk, as the key component of ABPP, can be used for peripheral nerve repair to yield better outcomes than ABPP.

In vitro callus induction and plantlet regeneration of Achyranthes aspera L.,a high value medicinal plant

Objective:To study callus induction from different explants(internode,leaf,root)and in vitro plantlets propagation from medicinally important plant Achyranthes aspera L.Methods:Sterilized explants were prepared by uning 0.1%HgCl_2 and 0.5%Bavistin and callus was obtained when cultured onto Murashige Skoog's(MS)medium by using different concentrations and combination of 2,4-D.NAA.BAP,IAA,IBA with 3%sucrose and 0.8%agar.Induced callus was immediately transferred to MS medium containing at different concentrations of phytohormones for shootlets and rootlets induction respectively.Results:Sterilization treatment of 0.1%HgCl_2.for 2-3 min and Bavistin 0.5%for 10-12 min showed the highest percentage of asepsis and survival rate.Maximum induction of callus was obtained from a combination of 2.0 mg/L 2,4-D and 0.5 mg/L NAA from leaf.Highest shootlets number(4.83±0.l7)and length(3.8±0.16)cm were observed on full strength MS medium when fortified with BAP 4.0 mg/L and KIN 0.5 mg/L.Concerted efforts of BAP 10 mg/L and NAA 0.5 mg/L on full strength MS medium showed highest leaf number(6.77±0.94).In vitro raised shoots were allowed to root on different strengths of MS medium fortified with IAA and IBA at different concentrations.Experimentally,3.0 mg/L IBA was enabled to induce maximum rootlets number(10.0±9.82)on full strength MS medium.Afterwards,regenerated shoots with well developed roots were successfully subjected to hardening process and were acclimatized.The survived plantlets showed 66.67%survival frequency without any morphological abnormality.Conclusions:The results demonstrated that different explants were good source of callus induction,morphology analysis as well as indirect plantlets regeneration.

In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants

Objective:To develop the reproducible in vitro propagation protocols for the medicinally important plants viz.,Achyranthes aspera(A.aspera)L.and Achyranthes bidentata(A.bidentata)Blume using nodal segments as explants.Methods:Young shoots of A.aspera and A.bidentata were harvested and washed with running tap water and treated with 0.1%bavistin and rinsed twice with distilled water.Then the explants were surface sterilized with 0.1%(w/v)HgCl_2 solutions for I min.After rinsing with sterile distilled water for 3-4 times,nodal segments were cut into smaller segments(1 cm)and used as the explants.The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog(MS)medium supplemented with 3%sucrose,0.6%(w/v)agar(HiMedia,Mumbai)and different concentration and combination of 6-benzyl amino purine(BAP),kinetin(Kin),naphthalene acetic acid(NAA)and indole acetic acid(IAA)for direct regeneration.Results:Adventitious proliferation was obtained from A.aspera and A.bidentata nodal segments inoculated on MS basal medium with 3%sucrose and augmented with BAP and Kin with varied frequency.MS medium augmented with 3.0 mg/L of BAP showed the highest percentage(93.60±0.71)of shootlets formation for A.aspera and(94.70±0.53)percentages for A.bidentata.Maximum number of shoots/explants(10.60±0.36)for A.aspera and(9.50±0.56)for A.bidentata was observed in MS medium fortified with 5.0 mg/L of BAP.For A.aspera,maximum mean length(5.50±0.34)of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A.bidentata(5.40±0.61)was observed in the very same concentration.The highest percentage,maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of 1BA.Seventy percentages of plants were successfully established in polycups.Sixty eight percentages of plants were well established in the green house condition.Sixty five percentages of plants were established in the field.Conclusions:The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A.aspera and A.bidentata.The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can he easily adopted for commercial large scale cultivation.

Gastroprotective effect of Achyranthes aspera Linn,leaf on rats

Objective:The study was aimed al evaluating the antiulcer activity of ethanolic extract of Achyranthes aspera(EEAA) leaf.Methods:The anti-ulcer assays were performed on pylorus ligation and chronic ethanol induced ulcer model.The effects of the EEAA on gastric content volume,pH.free acidity,total acidity and ulcer index were evaluated.Results:The percentage of ulcer protection(59.55%and 35.58%) was significantly(P 【 0.001) higher in the groups treated with the high dose of EEAA(600 mg/kg),it also reduced the volume of gastric juice and total acidity whereas,gastric pH was increased signiGcandy.Conclusions:The results of this study show significant gastroprotective activity of EEAA may be due to presence of phyto-constituents like flavanoids,saponins and tannins.

Chemical Components of Achyranthes bidentata Leaves by Ultra High Performance Liquid Chromatography-Mass Spectrometry

[Objectives] To study the chemical components and relative content of Achyranthes bidentata leaves and provide a scientific basis for further development and utilization of A. bidentata leaves.[Methods] The chemical components of A. bidentata leaves were rapidly analyzed using the ultra high performance liquid chromatography-time of flight-high resolution mass spectrometry (UHPLC-TOF-MS).[Results] Thirty eight chemical compounds were identified in samples of A. bidentata leaves collected from Wen County of Henan Province, in which seven chemical compounds had the relative content higher than 5%, linoleic acid reached 25.7% and inokosterone A reached 13.8%.[Conclusions] A. bidentata leaves contain many kinds of chemical compounds. This study is expected to provide a certain basis for further extraction of linoleic acid and inokosterone A.

One-pot synthesis of silver nanocomposites from Achyranthes aspera:An eco-friendly larvicide against Aedes aegypti L.

Objective:To formulate silver nanocomposites from Achyranthes aspera leaf extracts and evaluate its larvicidal activity against Aedes aegypti.Methods:The silver nanocomposites were synthesized from Achyranthes aspera leaf extracts.The process was optimized and traced through UV-visible and photon correlation spectroscopy.The larvicidal potential of silver nanocomposites of Achyranthes aspera leaf extracts was assessed against the early fourth instars of Aedes aegypti and three non-target organisms.Furthermore,the most effective and eco-safe nanocomposite was characterized by different biophysical techniques including scanning electron microscopy(SEM),energy dispersive X-ray(EDX)spectroscopy,transmission electron microscopy(TEM),X-ray diffraction(XRD)and Fourier transform-infrared spectroscopy(FT-IR).Results:The formulated silver nanocomposites exhibited efficient larvicidal efficacy against Aedes aegypti.Bioassay with silver nanocomposites formulated using different Ag NO3 concentrations(3,4,and 5 m M)revealed respective LC50 values of 37.570,6.262 and 1.041μg/m L;5.819,1.412 and 0.489μg/m L;and 5.519,1.302 and 0.267μg/m L after 24,48 and 72 h.The silver nanocomposites with 4 m M Ag NO3 were selected for characterization.SEM and TEM analysis revealed spherical,poly-dispersed structure with varied diameters of 1-25 nm.The XRD analysis established the crystalline and face-centred-cubic structure of silver nanocomposites with the maximum peak at a 2θvalue of 37.42°.The EDX pattern showed the presence of Ag,O and C in the nanocomposites in their order of weight%.The FT-IR displayed visibly distinct peaks in different ranges demonstrating the intricacy of silver nanocomposites.In addition,the lethal concentrations of silver nanocomposites of Achyranthes aspera leaf extracts against Aedes aegypti larvae were non-toxic to non-target organisms including Gambusia affinis,Daphnia magna and Moina macrocopa.Conclusions:Silver nanocomposites synthesized with leaf extract of Achyranthes aspera provide a cost-effective and eco-safe alternative to conventional insecticides,and can be utilized as a potent mosquito nano-larvicide.

Achyranthes bidentata polypeptide protects dopaminergic neurons from apoptosis induced by rotenone and 6-hydroxydopamine

It has been well documented that Achyranthes bidentata polypeptides（ABPPs） are potent neuroprotective agents in several types of neurons. However, whether ABPPs protect dopaminergic neurons from apoptosis induced by neurotoxins is still unknown. This study was designed to observe the effect of ABPPk, a purified fraction of ABPPs, on apoptosis of dopaminergic neurons. SH-5YHY cells and primary dopaminergic neurons were pre-treated with ABPPk（25, 50, or 100 ng/mL） for 12 hours. Cells were then exposed to 6-hydroxydopamine（50 or 150 μM） or rotenone（50 or 200 μM） for 36 hours to induce cell apoptosis. Our results demonstrate that ABPPk markedly increased viability in SH-SY5Y cells and primary dopaminergic neurons, decreased lactate dehydrogenase activity and number of apoptotic dopaminergic neurons, elevated mitochondrial membrane potential, and increased Bcl-2/Bax ratio. These findings suggest that ABPPk protects dopaminergic neurons from apoptosis, and that ABPPk treatment might be an effective intervention for treating dopaminergic neuronal loss associated with disorders such as Parkinson＇s disease.

Protective effects of Achyranthes bidentata polypeptides on retinal ganglion cells post-optic nerve crush in rats

Achyranthes bidentata polypeptides（ABPP） have been reported to inhibit apoptosis of retinal ganglion cells（RGCs）.The present study investigated the protective effects of ABPP on RGCs in a rat model of optic nerve injury.With prolonged injury time,RGC densities were gradually decreased.ABPP（5 μg） significantly increased RGC densities and upregulated growth associated protein 43 expression in rats with optic nerve injury.Results demonstrate that ABPP can protect RGCs and promote axonal growth after optic nerve crush.

Effects of Pb on Rhizosphere Soil Enzyme Activity and Chemical Constituents of Achyranthes bidentata Blume

[Objectives]The purpose of this study was to investigate the effects of Pb on rhizosphere soil enzyme activity and chemical constituents of Achyranthes bidentata Blume.[Methods]A.bidentata Blume plants were cultivated in self-made polyvinyl chloride(PVC)pots.The soil was added with different levels of Pb(0,200,400,600,800 and 1000 mg/kg air-dried soil)to investigate the effects of Pb on dry mass,active ingredients(oleanolic acid and ecdysterone)and rhizosphere soil enzyme activity of A.bidentata Blume.[Results]The root dry mass of A.bidentata Blume cultivated in the soil with Pb level above 400 mg/kg significantly reduced.The Pb residues in the A.bidentata Blume.plants growing in the soil with Pb level below 400 mg/kg complied with national standard.The contents of oleanolic acid and ecdysterone in A.bidentata Blume growing in the soil with Pb level above 600 mg/kg declined significantly.At the Pb level of 1000 mg/kg,the activity of urease was inhibited significantly.The activity of phosphatase was inhibited in the presence of Pb in the soil.The activity of sucrase was activated in the soil with Pb level below 400 mg/kg,and was inhibited in the soil with Pb level above 400 mg/kg.[Conclusions]This study has important guiding significance for the reasonable selection of planting base for A.bidentata Blume and the guarantee of its yield and quality.

Determination of Oleanolic Acid in Achyranthes aspera L. of Different Production Areas by HPLC

[Objectives] To determine the content of oleanolic acid in 10 batches of Achyranthes aspera L. from different production areas,and to establish a high performance liquid chromatography( HPLC) method for the determination of oleanolic acid in A. aspera. [Methods]Agilent C18 liquid chromatography column( 250 mm × 4. 6 mm,5 μm) was used for gradient elution with acetonitrile∶ water = 61∶ 39 as the mobile phase. The flow rate was 1 mL/min,the detection wavelength was 210 nm,and the column temperature was 35℃. [Results] The oleanolic acid injection volume showed a good linear relationship with the peak area in the range of 0. 382-7. 640 μg. The linear equation of oleanolic acid was: A = 530. 76 C,R = 1. 000 0; the range of sample recovery rate was 96. 03%-102. 73%,and the RSD value was 2. 16%( n =9). The content of the oleanolic acid was the highest in A. aspera produced in Guilin City of Guangxi( 0. 92%),and the lowest oleanolic acid content was in sample of Sitang in Nanning City of Guangxi( 0. 27%). It is recommended that the oleanolic acid content of A. aspera should not be lower than 0. 20%. [Conclusions]The HPLC method is accurate,reliable,easy to operate,and has good resolution. It is suitable for the determination of the content of oleic acid in A. aspera. It can provide reference for the quality control and standard drafting of A. aspera.

Interstitial lung disease induced by the roots of Achyranthes japonica Nakai:Three case reports

BACKGROUND The roots of Achyranthes japonica Nakai(AJN),called“Useul-puli,”has been traditionally used to control pain and improve dysfunction in osteoarthritis patients in South Korea.CASE SUMMARY We described 3 patients diagnosed with herbal medicine induced interstitial lung disease after consuming boiled the roots of AJN.They were referred to our hospital because of the modified Medical Research Council grade 4 dyspnea.Chest computed tomography showed bilateral ground-glass opacities with patchy consolidation.After treatment with systemic glucocorticoid therapy and discontinuation of the roots of AJN,their symptoms improved,and almost all ground-glass opacities and patchy consolidations on chest radiography and chest computed tomography resolved.CONCLUSION We present three cases of interstitial lung disease induced by the roots of AJN.

Deep chemical identification of phytoecdysteroids in Achyranthes bidentata Blume by UHPLC coupled with linear ion trap-Orbitrap mass spectrometry and targeted isolation

Achyranthes bidentata Blume is widely used as a traditional Chinese medicine with the effects of nourishing the liver and kidneys and strengthening muscles and bones.In this work,a rapid and simple strategy was developed for characterizing phytoecdysteroids by ultra-high-performance liquid chromatography coupled with liner ion trap-Orbitrap mass spectrometry using electrospray ionization in the negative mode.As a result,47 phytoecdysteroids were unambiguously or tentatively characterized.Among them,seven known compounds were identified according to the reference standards along with molecular formula,retention time and fragmentation patterns,while others were mostly potential new compounds.Through targeted isolation,the structures of three new compounds were determined by NMR spectra,which were consistent with LC-MS characterization.The present study provides an efficient method to deeply characterize phytoecdysteroids.

Effect of Hydro-ethanolic (1:1) Extract of Stephania Hernandifolia and Achyranthes Aspera in Composite Manner on Testicular Activity in Male Albino Rat:A Dose Dependent Analysis

Objective To investigate the effect of hydro-ethanolic extraet of Stephania hernandifolia leaves and Aehyranthes aspera roots in composite manner at the ratio of 1：3 on testicular activity in male rats. Methods Rats were divided into 4 groups with 8 animals in each group. The control （group A） received 0. 5 ml of olive oil/100 g body weight orally, other three groups were treated with said extract orally at a dose of 0.4 mg/g （group B） or 0.8 mg/g （group C） or 1.6 mg/g body weight （group D） respectively for 28 d. On 29th day of experiment, the animals were sacrificed. Sperm concentrations in cauda epididymis and biochemical markers like testicular cholesterol, androgenic key enzyme activity, plasma testosterone level seminal fructose level, glutamate oxaloacetate transaminase （GOT） and glutamate pyruvate transaminase （GPT） activities and serum triglyceride levels were measured following standard methods. ResuIts Extract treated animals in all doses resulted in a significant （P〈O. 05）decrease in sperm concentration, testicular androgenic key enzyme activity, plasma testosterone and seminal vesicle fructose levels along with an increase in testieular cholesterol level Animals treated at a dose of O. 8 mg/g body weight showed more promising result without causing any metabolic toxicity compared with other doses. Histological study also supported the biochemical results. The minimum but most effective dose i.e. 0.8 mg/g body weight had an inhibitory effect on implantation focused here by mating experiment.Conclusion The composite efficacy as male contraceptive extract of S. hernandifolia and A. aspera has a potent that may provide clues to the pharmaceutical industries for male contraceptive development

Studies on Synthesis and Antitumor Activity of Phosphorylated Achyranthes bidentata Polysaccharide (P-AbPS)

The synthesis of phosphorylated Achyranthes bidentata polysaccharide (P-AbPS) was reported based on different strategies. The P-AbPS with high degree of substitution (D.S. > 0.5) was obtained when phosphorus oxychloride (POCl3) was used as a phosphorylating agent and trimethyl phosphate-pyridine or dimethyl formamide was used as solvent. The influences of different solvents and reaction conditions were discussed. The pharmacology assay shows that P-AbPS possesses antitumor activity against sarcoma 180 and Lewis lung cancer in mice.

怀牛膝的三萜皂苷成分研究

目的对怀牛膝(AchyranthesebidentataBl.)根中三萜皂苷类成分进行研究,阐明其药效物质基础。方法利用硅胶柱、ODS柱、制备液相等进行分离,利用光谱技术和化学常数鉴定化合物的结构。结果分离并鉴定了7个化合物:即齐墩果酸(Ⅰ),竹节参皂苷-1(Ⅱ),齐墩果酸-3-O-β-D-吡喃葡萄糖醛酸苷(Ⅲ),齐墩果酸-3-O-β-D-(6′-丁酯)-吡喃葡萄糖醛酸苷(Ⅳ),齐墩果酸-3-O-β-D-(6′-甲酯)-吡喃葡萄糖醛酸苷(Ⅴ),3-O-(β-D-吡喃葡萄糖醛酸)-齐墩果酸-28-O-(β-D-吡喃葡萄糖)(Ⅵ),chikusetsusaponinⅣaethylester(Ⅶ)。结论化合物Ⅳ,Ⅴ,Ⅶ为首次从该植物中分离得到。