Comprehensive evaluation of the quality of Eriobotrya japonica leaves from different producing areas based on entropy weight TOPSIS method / 中国药房

OBJECTIVE To comprehensively evaluate the quality of Eriobotrya japonica leaves from different producing areas. METHODS The contents of alcohol-soluble extracts were determined by hot-dipping method using 30 batches of E. japonica leaves from different producing areas as samples. The contents of total flavonoids and total triterpene acids were determined by ultraviolet spectrophotometry. The contents of five kinds of triterpenic acids （euscaphic acid，crataegolic acid，corosolic acid，oleanolic acid and ursolic acid） were determined by HPLC. The quality of E. japonica leaves from different producing areas was comprehensively evaluated by using entropy weight technique for order preference by similarity to an ideal solution （TOPSIS）. The bivariate correlation analysis of E. japonica leaves was conducted by SPSS 22.0 software in terms of weight， comprehensive evaluation value， the content of alcohol-soluble extract， the contents of total flavonoids， total triterpene acids and five triterpenic acids. RESULTS The contents of alcohol-soluble extract in 30 batches of E. japonica leaves were （24.56±0.08）%-（34.85±0.13）%； the contents of total flavonoids were （4.69±0.11）-（14.23±0.27） mg/g； the contents of total triterpene acid were （27.58±0.59）- （63.95±1.27） mg/g； the contents of euscaphic acid， crataegolic acid， corosolic acid， oleanolic acid and ursolic acid were （0.728± 0.011）-（6.064±0.063）， （0.526±0.013）-（3.245±0.022）， （1.222±0.025）-（8.807±0.094）， （0.856±0.021）-（2.931±0.075）， （4.704±0.087）-（11.806±0.283） mg/g， respectively. The analysis result of entropy weight TOPSIS method showed that the top three samples with comprehensive evaluation values （No.Kjcx-5） were S14 （Huotian Town， Yunxiao County， Zhangzhou，Fujian）， S19 （Qinnan District， Qinzhou， Guangxi） and S29 （Guoyang County， Bozhou， Anhui）. Comprehensive evaluation 0596-2559522。E-mail：jxrcwxp@163.com of E. japonica leaves was positively correlated with the contents of five kinds of triterpenic acids， such as euscaphic acid， crataegolic acid， corosolic acid， oleanolic acid and ursolic acid （P＜0.01）. The weight of E. japonica leaves was positively correlated with the comprehensive evaluation value （P＜0.01）. CONCLUSIONS The qualities of E. japonica leaves from different producing areas are very different. Among them， the qualities of E. japonica leaves from Huotian Town， Yunxiao County， Zhangzhou of Fujian， Qinzhou Qinnan District of Guangxi， and Bozhou Guoyang County of Anhui are relatively better. The weight of E. japonica leaves is positively correlated with their quality.

Two new 2,3-seco triterpenoids from Rubus pirifolius Smith / 药学学报

Eight compounds were isolated from the ethyl acetate fraction of the 80% aqueous ethanol extract of the roots and stems of Rubus pirifolius Smith by AB-8 macroporous resin, silica gel, ODS, Sephadex LH-20 column chromatography, and semi-preparative HPLC. Their structures were identified by spectral analysis such as 1D/2D NMR, MS, UV, IR and by comparison with literature information as rubussecotriterpene A (1), rubussecotriterpene B (2), cecropiacic acid (3), cecropiacic acid 3-methyl ester (4), alphitolic acid (5), betulinic acid (6), betulin (7), and obtusalin (8). Compounds 1 and 2 are new compounds, and compounds 3-8 were isolated from this plant for the first time.

Quality evaluation of crude drug and different processed products of Eriobotryae Folium / 中国药房

OBJECTIVE To evalu ate the quality of crude drug and di fferent processed products of Eriobotryae Folium . METHODS Ten batches of Eriobotryae Folium were processed into honey-stir-baked Eriobotryae Folium ，ginger-juice-stir-baked Eriobotryae Folium ，ginger-juice-boiled Eriobotryae Folium ，licorice-juice-stir-baked Eriobotryae Folium ，licorice-juice-boiled Eriobotryae Folium ，stir-fried Eriobotryae Folium ，totally 70 batches of samples . The contents of alcohol-soluble extracts ，the contents of total triterpene acids （calculated by ursolic acid ）and five triterpene acids such as euscaphic acid were determined by hot-dipping method ，ultraviolet and visibe spectrophotometry and high performance liquid chromatography （HPLC），respectively. The fingerprints were established with HPLC and their similarity evaluation was conducted with Similarity Evaluation System of TCM Chromatographic Fingerprint （2004A）. Common peaks were identified by comparison with mixed control. Hierarchical clustering analysis ，principal component analysis and orthogonal partial least squares-discriminant analysis （OPLS-DA） were performed by using SPSS 22.0 software and SIMCA-P 14.1 software. RESULTS In Eriobotryae Folium ，honey-stir-baked Eriobotryae Folium ，ginger-juice-stir-baked Eriobotryae Folium ，ginger-juice-boiled Eriobotryae Folium ，licorice-juice-stir-baked Eriobotryae Folium ，licorice-juice-boiled Eriobotryae Folium ，stir-fried Eriobotryae Folium ，average contents of alcohol-soluble extracts were 25.90％，39.95％，27.44％，28.20％，28.38％，26.36％ and 29.26％；average contents of total triterpene acids were 40.62，49.33，52.56，46.38，52.17，55.06 and 53.41 mg/g；average contents of euscaphic acid ，crataegolic acid ，corosolic acid ， oleanolic acid ，ursolic acid and average total content were 1.966-4.808，1.459-2.824，4.525-8.172，1.294-1.817，6.294-8.470， 15.538-25.671 mg/g，respectively. There were 11 common peaks in 70 batches of samples ，and the peak 2，5，6，10 and 11 were identified as euscaphic acid ，crataegolic acid ，corosolic acid ， oleanolic acid and ursolic acid. The similarities of crude drug different processed products with crude drug fringer print were 0.919-1.000. Among 70 batches of samples ，10 batches of Eriobotryae Folium could be clustered into one category ，and 10 batches of ginger- juice-boiled Eriobotryae Folium could be clustered into one category ；other 50 batches of processed products of Eriobotryae Folium could be clustered into one category ； the cumulative variance contribution rate of the first two principal components was 80.682％；variable importance in projection （VIP）value was in descending order ，i.e. peak 2（euscaphic acid ）＞peak 5（crataegolic acid ）＞peak 6（corosolic acid ）＞peak 9 （unknown component ） ＞peak 11 （ursolic acid ）＞peak 10 （oleanolic acid ）， which of them were all higher than 1. CONCLUSIONS After processing ，the contents of alcohol-soluble extracts ，total triterpene acids and the total content of five triterpene acids （euscaphic acid ，crataegolic acid ，corosolic acid ，oleanolic acid and ursolic acid ）increased in varying degrees ， among which the content of alcohol-soluble extracts in honey-stir-baked Eriobotryae Folium was the highest ，the content of total triterpene acids in licorice-juice-boiled Eriobotryae Folium was the highest ，and total content of five triterpene acids in ginger- juice-boiled Eriobotryae Folium was the highest. Euscaphic acid ，crataegolic acid ，corosolic acid ，ursolic acid ，oleanolic acid and other components may be the differential components affecting the quality of raw and processed the leaves from Eriobotryae Folium .

Antiglycation, antioxidant, antiacne, and photoprotective activities of crude extracts and triterpene saponin fraction of Sapindus saponaria L. fruits: An in vitro study

Objective: To evaluate the photoprotective, antioxidant, antiglycation, and antiacne activities of crude extract (CESs) and triterpene saponin fraction (TSSs) of Sapindus saponaria. Methods: HPLC-MS purification was performed on a Symmetry TM C18 column. The saponins were identified by a UV detector. Antioxidant activity was evaluated by DPPH and O 2 - radicals scavenging, and FRAP and TBARS assays. Glycation activity was assessed by relative electrophoretic mobility and inhibition of advanced glycation end products (AGEs) formation. Additionally, antiacne activity was determined by inhibition of Cutibacterium acnes, and photoprotective effect was evaluated by Mansur's method. Results: Most of the triterpene saponins detected in the fraction by HPLC-MS analysis were hederagenin as the aglycon. CESs and TSSs presented varying antioxidant activity in DPPH (CESs: 75.69% and TSSs: 83.65%), FRAP (CESs: 425.39 μM TE/g DW and TSSs: 649.36 μM TE/g DW), TBARS (CESs: 42.96% and TSSs: 52.16%) and O 2 - radicals scavenging (CESs: 61.33% and TSSs: 86.69%) tests. CESs and TSSs also exhibited antiglycation activity comparable to bovine serum albumin treated with aminoguanidine. In addition, CESs and TSSs showed inhibition of AGE formation (34.48% and 61.85%, respectively). Antiacne activity against Cutibacterium acnes was observed with a minimum inhibitory concentration equal to minimum bactericidal concentration (CESs: 36.11 μg/mL and TSSs: 18.34 μg/mL). In photoprotective assays, CESs and TSSs showed maximum absorbance of 1.42 to 0.20 and 2.80 to 1.30, respectively, in the wavelength range of 260 to 400 nm. Furthermore, CESs and TSSs showed sun protection factors of 8.89 and 14.89, respectively. Conclusions: Sapindus saponaria fruit extracts show strong antioxidant potential and antiglycation activity against bovine serum albumin glycation and AGE formation. Besides, they presented antibacterial activity against Cutibacterium acnes and photoprotective effect against UV-A and UV-B.

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 .

A new tirucallane-type triterpene from the gum resin of Boswellia carterii / 药学学报

Two compounds were isolated from 95% ethanol extract of the gum resin of Boswellia carterii by silica gel column chromatography (CC) and high-performance liquid chromatography (HPLC). Their structures were identified with IR, UV, NMR and HR-ESI-MS spectroscopic data as 7α-hydroxy-3,11-dioxo- tirucalla-8,24-dien-21-oic acid (1) and 21β-hydroxy-3-acetyl-11-keto-β-boswellic acid (2). In addition, their absolute configurations were also identified by ECD calculations. Among them, compound 1 is a new compound and the absolute configuration of compound 2 is confirmed by ECD calculation for the first time.

Self-assembly performance of triterpene natural small molecules and their application in synergistic antitumor chemotherapy / 药学学报

Natural biocompatible nanomaterials such as self-assembled triterpene natural small molecule products with favorable anticancer activity show great potential for biomedical application. However, the mechanisms of their molecular self-assembled structures have not been investigated systematically, while there are still few reports of the natural active carrier for drug delivery. Herein, in this work, we further explored the molecular assembly mechanism and common regularity of tetracyclic triterpenes ergosterol, stigmasterol as well as pentacyclic triterpenes glycyrrhetinic acid and ursolic acid, which suggested that the coplanarity and orderliness of molecular arrangements which are speculated to be responsible for their self-assembly into the spherical, rod-like or lamellar nanostructure. Besides, ergosterol (ET) with better anticancer activity was chosen as a representative substance for construction of the synergistic antitumor nanodrug. By intermolecular hydrogen bonding and π-π stacking, chemotherapeutic drug paclitaxel (PTX) was encapsulated into ET-PTX NPs successfully. Then, the anti-cancer efficacy of the tumor-bearing mice was evaluated according to the protocol approved by the Experimental Animal Research Center of Harbin Medical University. The resulting nanodrug exhibited excellent biosafety and enhanced in vivo anticancer activity efficiency of 52.3%, higher than free PTX (29.4%) or ET NPs (32.5%) alone, further verifying the potential medical application value of triterpene natural products. This work provides not only a theoretical basis for exploring the self-assembly behavior of small molecule natural products, but also a promising perspective for the fabrication of active natural biocompatible nanodrug delivery systems for synergistic antitumor therapy and other biomedical applications.

Research progress on pharmacological activities of triterpenes from Ganoderma lucidum / 中国药理学通报

Ganoderma lucidum is a valuable traditional Chinese medicine with dual use of medicine and food in China. Its chemical components mainly include triterpenoids, polysaccharides, organic acids, alkaloids, amino acids and so forth. The triterpenoids mainly include ganoderma acid and ganoderma alcohol. This paper has summarized the pharmacological activities of Ganoderma lucidum triterpenes in anti-tumor, anti-inflammatory, liver and kidney protection, immune regulation, blood lipid and blood glucose lowering, and antimicrobial activities in recent years, in order to provide reference basis for the core efficacy e-valuation of high-quality Ganoderma lucidum. It also provides scientific evidence for the application of Ganoderma lucidum in health food and clinical medicine.

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.

Qualitative Analysis on Perilla frutescens Leaves and Stalks by UPLC-Q-Exactive-Orbitrap-MS / 中国实验方剂学杂志

Objective:To establish a method for qualitative analysis of components in Perilla frutescens leaves and stalks by liquid chromatography-mass spectrometry (LC-MS)，so as to explore the substance basis of pharmacodynamics differences between P.frutescens leaves and stalks.Method:P. frutescens leaves and stalks were extracted by 80% methanol-water ultrasound. The samples were analyzed by UPLC-Q-Exactive-Orbitrap-MS comprehensively. Halo-C18 column (2.1 mm×100 mm，2.7 μm) was used for gradient elution with 0.05% formic acid aqueous-0.05% acetonitrile formate as mobile phase in positive and negative ion modes. The flow rate was 0.3 mL·min-1，the column temperature was 40 ℃，and the injection volume was 5 μL.Result:The chemical compound in P. frutescens was deduced and identified based on the retention time of chromatography，and the exact molecular weight，excimer ion peaks，fragment ions and reference materials in Xcalibur software. The chemical composition of P. frutescens was identified by Mass Frontier 7.0 software. Totally 4 amino acids，7 phenylpropanoids，10 flavonoids，12 triterpenoids，7 organic acids，4 fatty acids，10 unknown compounds and 54 compounds were identified. Among them，6 triterpene acids, including glochidone, were identified in P. frutescens for the first time. The structures of five characteristic compounds were analyzed. There were 45 constituents in P.frutescens leaves and 32 constituents in P. frutescens stalks. They had 23 common constituents.Conclusion:LC-MS can identify the components of P. frutescens rapidly and effectively. This study provides an important theoretical basis for the quality control of different parts of P. frutescens and the development and utilization of P. frutescens.

A new triterpenoid saponin from leaves of Panax japonicas var. major / 中草药

Objective: To study the chemical composition from the leaves of Panax japonicus var. major. Methods: Column chromatographies (including macroporous resin, silica gel, Sephadex LH-20 and ODS) and semi-preparative HPLC were used to separate the constituents. The structures were elucidated by the analysis of spectral data and chemical properties. Results: Three compounds were isolated and elucidated as dammar-20(21),24-diene-3β,6α,12β-triol (1), dammar-20(22) Z,24-diene-3β,6α,12β-triol (2), and 3-O-[-β-D-glucopyranosyl-(1→2)-β-D-glucopyranosyl]-20-O-[-β-D-xylopyranosyl-(1→6)-β-D-glucopyranosyl]-23E,25-diene- 20(S)-protopanoxadiol (3). Conclusion: Compound 1 and 2 was obtained from the plant for the first time. And compound 3 named as majoroside Z is a new triterpenoid saponin.

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.

Cloning and bioinformatic analysis of β-AS gene from Panax japonicus / 中草药

Objective: The key enzyme of triterpene saponin metabolism was cloned and its sequence, structure and function were analyzed by bioinformatics. Methods: RNAs were extracted from the leaves of Panax japonicus The full-length cDNA sequences of β-AS were cloned by utilizing RT-PCR method, and the sequence was connected to the pMDTM18-T for cloning and sequencing.β-AS protein characteristics in transplanted species and cultivated species of P. japonicus were predicted and compared by bioinformatics analysis and the phylogenetic tree of β-AS protein was constructed. Results: The β-AS sequences in transplanted species and cultivated species of P. japonicus were obtained, which had 2 286 bp ORF and encoded 761 amino acids. There were little differences between the two varieties of β-AS proteins in physicochemical properties, secondary structure, tertiary structure, and phosphorylation sites, which may lead to show difference in catalytic activity. Conclusion: This work also obtained the full-length of cDNA sequence of β-AS gene in transplanted and cultivated varieties of P. japonicus, and provided a systemic sequence analysis of β-AS proteins, which can provide the useful information for β-AS studies in the future.

Anti-inflammatory effect of impressic acid from Acanthopanax gracilistylus based on NF-κB signaling pathway / 中草药

Objective: To explore the anti-inflammatory effect of impressic acid (IA) from Acanthopanax gracilistylus on lipopolysaccharide (LPS)-induced RAW264.7 cells. Methods: RAW264.7 cells were stimulated by LPS to establish an inflammatory model. The cytotoxicity of IA on RAW 264.7 cells was detected by EZ4U cell proliferation and cytotoxicity analysis kit. The level of nitric oxide (NO) was determined by Griess. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were determined by ELISA. The expressions of TNF-α and IL-1β mRNA were detected by RT-PCR. The expression of high-mobility group box 1 protein (HMGB1) was detected by western blotting. The levels of nuclear factor-κB (NF-κB) in cytoplasm and nucleus were measured by ELISA. Results: IA significantly suppressed the levels of TNF-α and IL-1β, the expression of HMGB1 protein, and the translocation of NF-κB from cytoplasm to nucleus in LPS-induced RAW264.7 cells (P < 0.05, 0.01). Conclusion IA from A. gracilistylus has an anti-inflammatory effect on LPS-induced RAW264.7 cells.

Brusatol: A potential anti-tumor quassinoid from Brucea javanica / 中草药·英文版

Brusatol, a triterpene lactone compound mainly from Brucea javanica, sensitizes a broad spectrum of cancer cells. It is known as a specific inhibitor of nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. In this review, we provide a comprehensive overview on the antitumor effect and molecular mechanisms of brusatol in vitro and in vivo. This review also covers pharmacokinetics studies, modification of dosages forms of brusatol. Increasing evidences have validated the value of brusatol as a chemotherapeutic agent in cancers, which may contribute to drug development and clinical application.

Application of Ganoderma lucidum and Ganoderma sinense Extract Reference Substance in the Fingerprint Analysis of Ganoderma / 中国药学杂志

OBJECTIVE: To carry out HPTLC and HPLC fingerprint analysis of 18 batches of Ganoderma samples using two kinds of reference substance of Ganoderma extract, G. lucidum Extract Reference Substance(CZERS) and G. sinense Extract Reference Substance(ZZERS). METHODS: HPTLC Fingerprint was used to analyze triterpene acids and sterols in Ganoderma with chloroform-acetonitrile-methanol-formic acid (13∶2∶0.5∶0.5, develop 3 times) and cyclohexane-ethyl acetate-methanol-formic acid (15∶5∶0.5∶0.5, develop 2 times) respectively. HPLC Fingerprint analysis was conducted using Kromasil 100-5 C18 column (4.6 mm×250 mm, 5 μm) kept at 25 ℃. Mobile phase A was acetonitrile and B was 0.02% phosphoric acid; gradient elution procedure was as follows: 0-40 min, 29%→33% A; 40-70 min, 33%→65%A; 70-105 min, 65%→100%A; 105-120 min, 100% A; flow rate was 1.0 mL•min-1. DAD detector was adopted with detection wavelength set at 244 nm. The injection volume was 10 μL. RESULTS: By using ERS and fingerprint analysis, G. lucidum, G. sessile and G. lucidum could be distinguished. The components of G. lucidum in different species and growth patterns were different. CONCLUSION: There are many varieties of G. lucidum, which can be divided into wild and artificial cultures, and the culture media of artificial culture are different, which leads to the difference of individual components of different G. lucidum. Fingerprint analysis based on ERS of specific varieties are more suitable for the overall quality control of G. lucidum.

Content Determination of 6 Kinds of Triterpene Acid in Tibetan Medicine Rubus biflorus by Pre-column Derivatization HPLC/ FLD-APCI/MS / 中国药房

OBJECTIVE： To establish the method for content determination of 6 kinds of triterpene acids such as haw acid， corosolic acid， betula acid， betulonic acid， oleanolic acid and ursolic acid in Tibetan medicine Rubus biflorus. METHODS： Pre-column devrivatization HPLC-FLD-APCI/MS method was adopted. 2-（7H-dibenzo[a，g]carbazol-7-yl） ethyl-4-methylbenzene- sulfonate was used as the pre-column derivatization reagent．Hypersil C18 column was used with the mobile phase consisted of 5％ acetonitrile water solution-acetonitrile （gradient elution） at the flow rate of 1.0 mL/min. The excitation wavelength of fluorescence was 300 nm and the emission wavelength was 395 nm. The column temperature was 35 ℃， and sample size was 10 μL. Under atmospheric-pressure chemical ionization （APCI） source in positive-ion mode， pressure was 60 psi， the drying gas flow rate was 9 L/min， the dry gas temperature was 350 ℃， the gasification temperature was 450 ℃， and the capillary voltage was 3 500 V. RESULTS： The linear range of haw acid，corosolic acid，betula acid，betulonic acid，oleanolic acid and ursolic acid were 0.025-6.4 μg/mL（r≥0.999 6）. The quantitative limits were 5.11， 4.78， 4.42， 4.22， 4.29， 4.51 ng/mL； and detection limits were 1.42， 1.27， 1.30， 1.28， 1.16， 1.22 ng/mL， respectively. RSD of precision test was less than 5％， stability and repeatability tests were all less than 2％（no betulonic acid detected）. The recovery rates were 97.90％-100.55％（RSD＝1.00％，n＝6）， 97.95％-102.95％（RSD＝1.74％，n＝6）， 96.00％-101.20％（RSD＝2.00％，n＝6）， 93.25％-104.20％（RSD＝4.25％，n＝6）， 92.20％-103.30％（RSD＝3.58％，n＝6）， 97.80％-103.50％（RSD＝2.03％，n＝6）， respectively. CONCLUSIONS： The method is accurate， reliable and exclusive， and can be used for simultaneous determination of 6 kinds of triterpene acids in Tibetan medicine R. biflorus.

Evaluation of chemical consistency of triterpene acids in ethanol extracts of Poria and acetic ether extracts thereof by UPLC-QTOF-MS/MS with full scan and mimic MRM mode / 药学学报

An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) method was developed to evaluate the chemical consistency of triterpene acids in ethanol extracts of Poria and acetic ether extracts thereof. First, high resolution mass spectrometry data were obtained with Full scan mode, by comparing with MS data from the reference compounds and literatures, a total of 23 components were unequivocally or tentatively identified in ethanol extracts and acetic ether extracts thereof. Then, a mimic multiple reaction monitoring (mMRM) mode was established using UPLC-QTOF-MS/MS to quantify the triterpene acids in ethanol extracts and acetic ether extracts thereof. Eleven components were absolutely quantified with reference compounds, while 12 components without reference compounds were relatively quantified with peak areas, the transfer and enrichment rate of triterpene acids during liquid-liquid extraction were calculated. It was found all of the 23 triterpene acids identified in Poria ethanol extracts could be transferred into acetic ether extracts with high transfer and enrichment rate. The present study provides not only scientific evidence for further extraction of triterpene acids in Poria by acetic ether, but also an approach for comprehensive evaluation of the chemical consistency of herbal medicine extracts before and after the liquid-liquid extraction.

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.

A new triterpenoid and a new steroid from roots of Vladimiria souliei / 中草药

Objective: To study the chemical compositions from the roots of Vladimiria souliei. Methods: The ethanol extract of the roots of V. souliei was isolated by silica gel, Sephadex LH-20, MCI, HPLC, and C18 reverse-phase column chromatographies. Their physical and chemical structures were elucidated by analysis of NMR and MS spectroscopic data, and the absolute configuration of compound 1 was confirmed by Cu-Kα X-ray crystallographic analysis. Results: Three compounds were isolated from the roots of V. souliei, and named as 25-hydroxy-uvaol (1), vlasoudiol (2), and stigmasterol (3), respectively. Conclusion: Compounds 1 and 2 are identified as new compounds, and compound 3 is isolated from the roots of V. souliei for the first time.