Purification and Structural Characterization of Polysaccharides from Polygonum multiflorum Thunb. and Their Immunostimulatory Activity in RAW264.7 Cells.

Polygonum multiflorum Thunb. (PM) and derived products are broadly utilized in Chinese traditional medicine. According to our previous research, PM mostly contains polysaccharides, which display a wide range of biological activities. Two water-soluble polysaccharides (PMPs-1 and PMPs-2) were obtained from PM by DEAE-Cellulose and Sephadex G-100 column chromatography. Colorimetry, HPGPC-MALLS-RID, HPLC-PDA, methylation, FT-IR, NMR, and SEM were used to characterize these polysaccharides. PMPs-1 and PMPs-2 had average molecular weights of 255.5 and 55.7 kDa, respectively. PMPs-1 consisted of Man, Glc, Gal, and Ara at 0.9:78.6:1.0:1.6 and was a glucan with → 4)-Glcp-(1 → as a backbone. Meanwhile, PMPs-2, an acidic polysaccharide, comprised Rha, GalA, Glc, Gal, and Ara at 3.2:20.3:2.7:1.0:8.3. PMPs-1 and PMPs-2 significantly improved the proliferation of RAW 264.7 cells and induced NO, TNF-α, and IL-6 release. This study reveals that these two polysaccharides can be explored as novel immunomodulators and provide a basis for further development of PM in food and pharmaceutical industries.

Pyrrolizidine alkaloids and health risk of three Boraginaceae used in TCM.

Objective: The aim of this study was to systematically explore the pyrrolizidine alkaloids (PAs) type, content and risk assessment in the three Boraginaceae used in TCM, involving Arnebia euchroma (AE), A. guttata (AG), and Lithospermum erythrorhizon (LE). Method: A UHPLC-MS/MS method was established to simultaneously determine eight pyrrolizidine alkaloids (PAs), namely intermedine, lycopsamine, intermedine N-oxide, lycopsamine N-oxide, 7-acetyllycopsamine, 7-acetyllycopsamine N-oxide, echimidine N-oxide, and echimidine in the three herbs. Based on these results, the risk assessment was explored using the routine margin of exposure (MOE) combined with relative potency (REP) for oral and external usage, respectively. Results and Conclusion: Imermedine and imermedine N-oxide were common components in the eight tested PAs. 7-acetyllycopsamine and its N-oxide were not detected in AE; echimidine and its N-oxide were not detected in AG; lycopsamine and its N-oxide, 7-acetyllycopsamine and its N-oxide were not detected in LE. The total contents of 8 PAs in 11 batches of AG was341.56-519.51 µg/g; the content in 15 batches of LE was 71.16-515.73 µg/g, and the content in 11 batches of AE was 23.35-207.13 µg/g. Based on these results, the risk assessment was explored using MOE combined with REP for oral and external usage, respectively. The findings of the risk assessment method of PAs based on MOE combined with the REP factor were consistent with the clinical toxicity results. As an oral herb, AE had low risk or no risk due to its low PA contents, and individual batches of LE were medium risk, while attention should be paid to their clinical use.AG was also low risk. The external use of the three Boraginaceae used in TCM was not associated with any risk. This study systematically explored the PA type and content of the three Boraginaceae used in TCM. Additionally, the refined risk assessment of PAs based on REP provided a more scientific basis for quality evaluation and rational use of the medicinal Boraginaceae used in TCM to improve public health.

Comparisons of physicochemical features and hepatoprotective potentials of unprocessed and processed polysaccharides from Polygonum multiflorum Thunb.

The raw and processed Polygonum multiflorum Thunb (PM) are used to treat different diseases, and PM has also been reported to have hepatotoxic effects. Moreover, mounting evidence indicates that processed PM is less toxic than raw PM. The changes in efficacy and toxicity of PM during the processing are closely related to the changes in chemical composition. Previous studies have mainly focused on the changes of anthraquinone and stilbene glycosides during process. Polysaccharides, as main components of PM, showed many pharmacological effects, but its changes in the processing has been neglected for a long time. In this study, the polysaccharides of PM in the raw (RPMPs) and processed products (PPMPs) were determined and the liver injury model induced by acetaminophen was utilized to evaluate the impact of polysaccharides on the liver. Results showed that the heteropolysaccharides RPMPs and PPMPs both comprised Man, Rha, GlcA, GalA, Glc, Ara and Xyl, but markedly differed in polysaccharide yield, molar ratio of monosaccharide composition and Mw. In vivo analysis, results showed that demonstrated that RPMPs and PPMPs both exerted hepatoprotective effects by upregulating antioxidant enzymes and repressing lipid peroxidation. It is noteworthy that the polysaccharide yield of processed PM was seven-fold higher than that of raw PM, so it is speculated that processed PM has better hepatoprotective effects at the same dose of decoction. The present work provides an important foundation for studying the polysaccharide activity of PM and further revealing the processing mechanism of PM. This study also proposed a new hypothesis that the significant increase of polysaccharide content in processed PM may be another reason that the product PM causes less liver injury.

[Determination of content of nine components in Xinnaojian preparations from different manufacturers by QAMS].

This study established the ultra-performance liquid chromatography(UPLC) fingerprint of Xinnaojian preparations. With epicatechin gallate as the internal reference substance, a quantitative analysis of multi-components by single marker(QAMS) method for determining the content of nine components(gallic acid, epigallocatechin, catechin, caffeine, epicatechin, epigallocatechin gallate, gallocatechin gallate, epicatechin gallate, and catechin gallate) in Xinnaojian preparations was established. The content determined by the external standard method(ESM) and QAMS method was compared to evaluate the feasibility and accuracy of QAMS method. The results showed that the standard curves of nine components had good linear relationship within the test concentration ranges. The average recoveries were 87.57%-107.4%, and the RSD was 1.5%-2.9%. Except epigallocatechin, the other components showed good repeatability under different experimental conditions. Epigallocatechin could meet the requirements in the same instrument and at the same wavelength. The results generally showed no significant difference between QAMS and ESM. The content of 9 components varied between the samples from different manufacturers, while it showed no significant difference between the samples from the same manufacturer. In summary, the UPLC fingerprint combined with QAMS method is feasible and accurate for determining the content of the nine components, which can be used for rapid quality evaluation of Xinnaojian preparations.

Integrative Metabolomics and Proteomics Detected Hepatotoxicity in Mice Associated with Alkaloids from Eupatorium fortunei Turcz.

The traditional Chinese herbal medicine Eupatorium fortunei Turcz. (E. fortunei) has been widely adopted to treat nausea, diabetes, siriasis, and poor appetite. However, E. fortunei contains multiple pyrrolizidine alkaloids (PAs). This study aimed to investigate the hepatotoxicity of total alkaloids in E. fortunei (EFTAs) and identify the toxic mechanisms of EFTAs on hepatocytes. Liquid chromatography with a tandem mass spectrometry assay with reference standards indicated that EFTAs mainly consisted of eight PAs whose content accounted for 92.38% of EFTAs. EFTAs markedly decreased mouse body and liver weights and increased the contents of AST and ALT. The histopathological assays demonstrated that, after exposition to EFTAs, the structures of hepatocytes were damaged and the fibrosis and apoptosis in hepatocytes were accelerated. Moreover, EFTAs increased the serum level of inflammatory cytokines and aggravated circulating oxidative stress. A combination of hepatic proteomics and metabolomics was used to investigate the toxic mechanisms of EFTAs. The study revealed that EFTAs seriously disrupted glycerophospholipid metabolism by upregulating the contents of lysophosphatidylglycerol acyltransferase 1 and phosphatidylinositol and downregulating the contents of choline/ethanolamine kinase beta, choline-ethanolamine phosphotransferase 1, phospholipase D4, 1-acylglycerophosphocholine, phosphatidylcholine, and dihydroxyacetone phosphate in the liver, resulting in detrimental inflammation, fibrosis, and apoptosis. This study revealed that EFTAs induced severe hepatotoxicity by disrupting glycerophospholipid metabolism.

Changes of Physicochemical Properties and Immunomodulatory Activity of Polysaccharides During Processing of Polygonum multiflorum Thunb.

The roots of Polygonum multiflorum Thunb (PM) have a long history of usage in traditional Chinese medicine and are still widely utilized today. PM in raw or processed form has different biological activities and is commonly used to treat different diseases. Polysaccharides are the main component of PM, and it is unclear whether their physicochemical properties and activities change after processing. In this study, the polysaccharides from thirty-one raw PM (RPMPs) and nine processed PM (PPMPs) were extracted, and the physicochemical properties and immunomodulatory activity in vitro of polysaccharide samples were evaluated. Results showed that RPMPs and PPMPs had significant differences in physicochemical properties. RPMPs and PPMPs were both composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, galactose, and arabinose. However, RPMPs and PPMPs had significant differences in their yields, molecular weight (Mw), and the molar ratio of Glc/GalA (p < 0.05), which can be used to distinguish raw and processed PM. The fingerprint of monosaccharide composition was analyzed by chemometrics, and it was further demonstrated that Glc and GalA could be used as differential markers. The immunomodulatory activity assays indicated that RPMPs and PPMPs could significantly enhance phagocytosis and mRNA expression of cytokines in RAW 264.7 cells. In addition, the immunomodulatory activity of PPMPs with lower Mw was significantly better than that of RPMPs. This study furthers the understanding of the polysaccharides from raw and processed PM and provides a reference for improving the quality standard of PM.

[Form and valence of arsenic in dry and fresh Cordyceps breeding products based on HPLC-ICP-MS and its risk assessment].

A comparative study was conducted for the first time on the form and valence of arsenic in the dry and fresh Cordyceps breeding products to clarify the specific morphology and valence of arsenic in Cordyceps breeding products and its safety. Arsenic betai-ne(AsB), arsenite(AsⅢ), dimethyl arsenic(DMA), arsenocholine(AsC), monomethyl arsenic(MMA), and arsenate(AsⅤ) in the dry and fresh samples were investigated using a bionic extraction method combined with HPLC-ICP-MS. The HPLC separation was performed on a DioncxIonPac~(TM) AS7 anion exchange column with a mobile phase of 100 mmol·L~(-1) ammonium carbonate-water for gradient elution at room temperature and the flow rate of 0.8 mL·min~(-1). HPLC was coupled with ICP-MS for the determination. The arsenic content was characterized in combination with chemometrics. The health safety risk of inorganic arsenic in the samples was assessed using the margin of exposure(MOE). The results of methodological validation showed that the six arsenic compounds showed good linearity(R~2>0.999) from 10 to 500 ng·mL~(-1), with precision RSDs of 1.8%-3.0%, recoveries(n=6) of 84.15%-98.28%, reproducibility RSDs of 6.4%-7.7%, and sample stability RSDs of 8.3%-14% within 10 h. Trace AsⅢ and AsⅤ were detected in 30 batches of dry and fresh Cordyceps breeding products, while arsenic compounds in other forms and valence were not detected. In the dry products, AsⅢ was 0.019-0.040 mg·kg~(-1) and AsV was 0.024-0.061 mg·kg~(-1), while in the fresh products, AsⅢ was 0.002 3-0.006 1 mg·kg~(-1) and AsⅤ was 0.008-0.016 mg·kg~(-1). The risk assessment results showed that the MOE of inorganic arsenic was much higher than 1 in both dry and fresh products, and the potential health safety risk of inorganic arsenic was low. The HPLC-ICP-MS method established in this study was efficient, rapid, accurate, and stable for the determination of six arsenic compounds in Cordyceps breeding products. The results of this study provide a basis for the safety and quality control of Cordyceps breeding products.

Neuroprotective Alkamides from the Aerial Parts of Achillea alpina L.

Three new alkamides, achilleamide B-D (1-3) along with five known alkamides (4-8) were isolated from the aerial parts of Achillea alpina L. Structures were elucidated by spectroscopic analysis. Modified Mosher's method and electronic circular dichroism (ECD) calculations were introduced for the absolute configuration of 3. The neuroprotective effects of all the compounds were evaluated by 6-hydroxydopamine (6-OHDA)-induced cell death in human neuroblastoma SH-SY5Y cells, with concentration for 50 % of maximal effect (EC50 ) values of 3.16-24.75 µM, and the structure-activity relationship was conducted.

Medicinal Earthworm: Speciation and Bioaccessibility of Arsenic and Its Potential Health Risks.

Arsenic in environmental health has caused public concerns all over the world. However, high levels of arsenic residues in medicinal animals have not received enough attention. Medicinal earthworms are consumed widely in China, but its arsenic potential health risks to humans are unknown. This work investigated the total concentration, bioaccessibility, and speciation of arsenic in earthworms by ICP-MS and HPLC-ICP-MS to evaluate its potential health risks to humans. Arsenic was found in all earthworms at concentrations ranging from 0.4 to 53.6 mg kg-1. The bioaccessibility of arsenic (bAs) varied significantly and ranged from 12.1 to 69.1%, with inorganic arsenic (iAs, including As(III) and As(V)) as the predominant species. Furthermore, a small amount of arsenobetaine (AB) was found. The estimated daily intake dose (EDI), hazard quotient (HQ), and carcinogenic risk (CR) of arsenic in most of the samples exceeded the safe threshold level. Results from this study indicated that the potential health risks by the consumption of earthworms may not be negligible. Herein, recommendations for the use of earthworms and regulatory recommendations for arsenic limit standards were proposed. This study reminds us that more control and monitoring of arsenic in medicinal animals should be carried out.

[Content determination of eight phenols in Ilex hainanensis harvested at different time by HPLC-DAD].

This study intends to develop a high performance liquid chromatography-diode array detection(HPLC-DAD) method for simultaneous determination of chlorogenic acid, 2-hydroxymethyl-3-hydroxyl-1-butene-4-O-ß-D-(6″-O-caffeoyl)-glucopyranoside, pubescenoside B, huazhongilexone-7-O-ß-D-glucopyranoside, rutin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C in Ilex hainanensis. The HPLC conditions are as follows: Waters XBridge C_(18 )column(4.6 mm×250 mm, 5 µm), mobile phase of 0.5% formic acid in water(A)-acetonitrile(B), gradient elution(0-8 min, 5%-12% B; 8-18 min, 12%-18% B; 18-30 min, 18%-25% B; 30-40 min, 25%-30% B; 40-42 min, 30%-80% B; 42-45 min, 80% B) at the flow rate of 0.8 mL·min~(-1), detection wavelengths of 282, 324, and 360 nm, column temperature of 25 ℃, and injection volume of 5 µL. The content of the 8 phenols in 8 samples was 0.30-6.29, 0.29-3.27, 0.15-10.4, 0.51-5.85, 0.49-9.02, 0.51-4.68, 1.93-13.4, and 0.87-5.95 mg·g~(-1), respectively. Moreover, the content of phenols in the samples collected in October was higher than that of samples harvested in other months. The established method is accurate and sensitive for the determination of phenols in I. hainanensis, which is useful for the quality improvement of this herbal medicine.

[Simultaneous determination of six pyrrolizidine alkaloids in different parts of Emilia sonchifolia by UPLC-MS/MS].

This study aims to develop a UPLC-MS/MS method for simultaneous determination of six pyrrolizidine alkaloids(PAs)--intermedine N-oxide(ImNO), lycopsamine N-oxide(LyNO), seneciphylline(Sp), seneciphylline N-oxide(SpNO), senecionine N-oxide(SnNO), and senkirkine(Sk) in different parts of Emilia sonchifolia. UPLC conditions are as follows: ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8 µm), mobile phase consisting of 0.05% formic acid and 2.5 mmol·L~(-1) ammonium formate in water(A)-0.05% formic acid and 2.5 mmol·L~(-1) ammonium formate in acetonitrile(B) for gradient elution. MS conditions are as below: electrospray ionization(ESI) in the positive ion mode, multiple reaction monitoring(MRM), and the content of the six PAs was calculated with the external standard method. The results suggested the differences in the six PAs among different parts of E. sonchifolia. Sk was detected in all the four parts, with similar content. SnNO also existed in all the four parts, but the content in roots was significantly higher than that in other parts. Sp and SpNO were found in both roots and flowers, with the content higher in the former than in the later. ImNO and LyNO were only found in leaves, and the content was low. Among the six components detected, ImNO, LyNO, and SpNO were found and determined for the first time, which enriched the toxic components and laid a scientific basis for the quality and safety evaluation of E. sonchifolia.

Simultaneous determination of eight pyrrolizidine alkaloids in various parts of Eupatorium lindleyanum by ultra high performance liquid chromatography tandem mass spectrometry and risk assessments based on a real-life exposure scenario.

Pyrrolizidine alkaloids are toxins having hepatotoxic and carcinogenic effects on human health. A ultra high performance liquid chromatography tandem mass spectrometry technique was developed for the first time for the simultaneous determination of eight pyrrolizidine alkaloids, including four diastereoisomers (intermedine, lycopsamine, rinderine, and echinatine) and their respective N-oxide forms, in different parts of Eupatorium lindleyanum. The risk assessment method for pyrrolizidine alkaloids in Eupatorium lindleyanum was explored using the margin of exposure strategy for the first time based on a real-life exposure scenario. Differences were found in all eight pyrrolizidine alkaloids in various parts of Eupatorium lindleyanum. Besides, the total levels of pyrrolizidine alkaloids in Eupatorium lindleyanum followed the order of root > flower > stem > leaf. Moreover, the risk assessment data revealed that the deleterious effects on human health were unlikely at exposure times of less than 200, 37, and 12 days during the lifetimes of Eupatorium lindleyanum leaves, stems, and flowers, respectively. This study reported both the contents of and risk associated with Eupatorium lindleyanum pyrrolizidine alkaloids. The comprehensive application of the novel ultra high performance liquid chromatography tandem mass spectrometry technique alongside the risk assessment approach provided a scientific basis for quality evaluation and rational utilization of toxic pyrrolizidine alkaloids in Eupatorium lindleyanum to improve public health safety.

Structural disorganization of cereal, tuber and bean starches in aqueous ionic liquid at room temperature: Role of starch granule surface structure.

The structural disorganization of different starches in a 1-ethyl-3-methylimidazolium acetate ([Emim][OAc])/water mixture (1:6 mol./mol.) at room temperature (25 °C) was studied. For normal cereal starches, which have pinholes randomly dispersed on the granule surface or only in the outermost annular region (wheat starch), the aqueous ionic liquid (IL) completely destroyed the granule structure within 1-1.5 h. Pea starch (PeS) granules with cracks were destroyed by the aqueous IL within 6 h. High-amylose maize starch (HAMS), as well as potato and purple yam starches (PoS and PYS), which have a dense and thick outer granule layer, were even more resistant to the action of the solvent. Structural disorganization was accompanied by increased viscosity and controlled the binding of water molecules with starch chains. From this study, we concluded that the surface characteristics of starch granule are an important factor affecting starch structural disorganization in an aqueous IL.

[Simultaneous determination of 6 main components in Achilleae Herba by HPLC].

This study aims to establish an HPLC method for the simultaneous determination of 6 main components, including chlorogenic acid, 3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid, pellitorine and neopellitorine B in Achil-leae Herba. HPLC analysis was performed on a Merck Purospher STAR RP-18 endcapped(4.6 mm×250 mm, 5 µm), with a mobile phase consisting of 0.05% phosphoric acid solution(A)-acetonitrile(B) at a flow rate of 1 mL·min~(-1)(0-7 min,12%-14% B;7-10 min,14%-17% B;10-25 min,17%-22% B;25-35 min,22%-35% B;35-51 min,35%-80% B;51-60 min,80%-90% B). The detection wavelength was 254 nm and the column temperature maintained at 30 ℃, and the injection volume was 5 µL. The standard curves revealed a good linear relationship. The contents of 6 components were 0.404%-2.116% for chlorogenic acid, 0.160%-0.892% for 3,4-dicaffeoylquinic acid, 0.608%-1.464% for 3,5-dicaffeoylquinic acid, 0.168%-0.868% for 4,5-dicaffeoylquinic acid, 0.122%-1.234% for pellitorine, 0.065%-0.312% for neopellitorine B, respectively. Both cluster and principal component analysis can distinguish the research data in anthesis and pre-anthesis by software Chempattern. There were obviously differences in the different harvest time. Therefore, attention should be paid to the harvesting time of the herb. The method can be used to determine the contents of six main components, and can provide reference for the improvement of quality standard of Achilleae Herba.

[HPLC specific chromatogram of Vernonia anthelmintica and determination of six components].

This work aims to establish an HPLC specific chromatogram and determine six components of Vernonia anthelmintica with chlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, scutellarein and luteolin as index components. HPLC analysis was performed on a Waters Xbridge C_(18) column(4.6 mm×250 mm, 5 µm) with gradient elution of acetonitrile-0.05% trifluoroacetic acid solution at a flow rate of 1.0 mL·min~(-1). The detection wave length was 360 nm and the column temperature was 30 ℃. Chemometrics software Chempattern was employed to analyze the data. HPLC specific chromatogram of V. anthelmintica was established and six characteristic peaks were marked. Six characteristic peaks were simultaneously determined by HPLC within 50 min. The contents of the six components in 13 batch samples of V. anthelmintica were 0.14%-0.68%, 0.44%-0.74%, 0.63%-1.01%, 0.14%-0.71%, 0.15%-0.26% and 0.010%-0.030%, respectively. The HPLC specific chromatogram of V. anthelmintica, together with determination of six components showed strong specificity, and it can be used for the quality control of the crude drug.

[Comparative studies of three Cistanche speices based on UPLC specific chromatogram and determination of main components].

Based on UPLC specific chromatogram and determination of seven main components,this study aimed at evaluating the quality of Cistanche deserticola,C. tubulosa and C. sinensis. Echinacoside,cistanoside A,verbascoside,tubuloside A,isoacteoside,2'-acetylacteoside,tubuloside B were used as reference substances. UPLC analysis was performed on a Waters ACQUITY UPLC HSS T3 column( 2. 1 mm×100 mm,1. 8 µm). The mobile phase was acetonitrile-0. 08% trifluoroacetic acid solution. The flow rate was0. 3 mL·min-1,and the injection amount was 10 µL. The column temperature was 40 ℃,and the detection wavelength was 330 nm.The UPLC specific chromatograms were processed with ChemPattern software. UPLC specific chromatograms of C. deserticola and C.tubulosa from different samples were of high similarity,but the similarities of their counterfeit C. sinensis were less than 0. 06. Both of cluster and principal component analysis can distinguish certified products and counterfeits. The content ratios of echinacoside/verbascoside and verbascoside/isoacteoside were quite different between C. deserticola and C. tubulosa,which had distinct significance.The UPLC specific chromatogram and contents of seven main components can provide a basis for quality evaluation of Cistanches Herba.

[Comparative study for freeze-drying and sun-drying on multi-index ingredients of Cordyceps sinensis].

The aim of this paper was to compare the influence of freeze-drying and sun-drying on six main nucleosides and nucleobases of Cordyceps sinensis by HPLC. Hypoxanthine,xanthine,uridine,inosine,guanosine and adenosine were reference substances. HPLC analysis was performed on a GL Inertsustain AQ-C_(18) column( 4. 6 mm×250 mm,5 µm),with mobile phase consisting of water( A)-acetonitrile( B) at the flow rate of 1. 0 mL·min~(-1)( 0-10 min,0-1% B; 10-65 min,1%-3% B). The detection wavelength was 260 nm,the column temperature was controlled at 30 ℃,and the injection volume was 5 µL. The linear ranges of hypoxanthine,xanthine,uridine,inosine,guanosine and adenosine were 1. 025-20. 50( r = 0. 999 8),0. 545-10. 90( r = 0. 999 9),4. 051-81. 02( r = 0. 999 8),4. 044-80. 88( r= 0. 999 9),2. 075-41. 50( r= 0. 999 9),4. 032-80. 64( r = 0. 999 9) mg·L~(-1),respectively. The average recoveries of them( n = 6)were as follows: 102. 3%( RSD 2. 1%),101. 1%( RSD 2. 4%),97. 80%( RSD 1. 7%),101. 8%( RSD 1. 8%),98. 90%( RSD2. 0%) and 98. 10%( RSD 1. 4%),respectively. Each sample was processed by freeze-drying and sun-drying so as to compare the difference between the two drying methods. The contents of six index ingredients were significantly different between freeze-drying and sun-drying sample of C. sinensis. The total contents of six index ingredients in sun-drying sample were higher than that in the corresponding freeze-drying sample. This research results provide the scientific basis for the drying methods and quality evaluation of C. sinensis.

Antibacterial triterpenoids from the leaves of Ilex hainanensis Merr.

One new triterpenoid (1) and seven known analogues (2-8) were isolated from the leaves of Ilex hainanensis Merr.. Their structures were established by analysis of their MS, 1D and 2D NMR spectroscopic data and comparison with those in the literature. The antibacterial activity of compounds 1-8 were evaluated by determination of minimum inhibitory concentration using twofold microdilution broth method against Streptococcus mutans ATCC 25175 (Gram-positive) and Fusobacterium nucleatum ATCC 10953 (Gram-negative). Compounds 3 and 5 showed significant antibacterial activity against S. mutans in concentration of 9.7 µg/mL, while showed little antibacterial activity against F. nucleatum. On the contrary, the inhibitory activity of compounds 1, 2 and 6 against F. nucleatum were higher than them against S. mutans.

[Guaianolides from aerial parts of Artemisia myriantha].

This study was performed to investigate the guaianolides from the aerial parts of Artemisia myriantha. The chemical constituents were isolated by chromatographic columns over silica gel, Sephadex LH-20, and ODS, as well as Semi-prep HPLC methods, and their structures were identified by NMR and MS data. Ten compounds were isolated and identified as follows: artemyriantholide E (1), tanaphillin (2), 1ß, 10ß-epoxydehydroleucodin (3), 5-hydroxyleucodin (4), dehydrocostuslactone (5), 3-O-methyl-iso-secotanapartholide (6), roxbughianin A (7), dehydroleucodin (8), arglabin (9), and 8α-acetoxyarglabin (10). Compound 1 was a new compound, and compounds 2-7 were isolated from this plant for the first time. Compound 3 exhibited selective cytotoxicity against human liver cancer (Bel-7402) with IC50 value of 5.35 µmol·L⁻¹, and 6 against human gastric cancer (BGC-823) with IC50 value of 2.68 µmol·L⁻¹, respectively.

[Study on ethnic medicine quantitative reference herb,Tibetan medicine fruits of Capsicum frutescens as a case].

High price and difficult to get of reference substance have become obstacles to HPLC assay of ethnic medicine. A new method based on quantitative reference herb (QRH) was proposed. Specific chromatograms in fruits of Capsicum frutescens were employed to determine peak positions, and HPLC quantitative reference herb was prepared from fruits of C. frutescens. The content of capsaicin and dihydrocapsaicin in the quantitative control herb was determined by HPLC. Eleven batches of fruits of C. frutescens were analyzed with quantitative reference herb and reference substance respectively. The results showed no difference. The present method is feasible for quality control of ethnic medicines and quantitative reference herb is suitable to replace reference substances in assay.