Co-reporter:Dong-Sheng Zhao, Li-Long Jiang, Ya-Xi Fan, Ling-Li Wang, Zhuo-Qing Li, Wei Shi, Ping Li, and Hui-Jun Li
Chemical Research in Toxicology October 16, 2017 Volume 30(Issue 10) pp:1865-1865
Publication Date(Web):September 13, 2017
DOI:10.1021/acs.chemrestox.7b00176
The use of herbal medicines continues to expand globally, meanwhile, herb-associated hepatotoxicity is becoming a safety issue. As a conventional Chinese medicinal herb, Dioscorea bulbifera rhizome (DBR) has been documented to cause hepatic toxicity. However, the exact underlying mechanism remains largely unexplored. In the present study, we aimed to profile entire endogenous metabolites in a biological system using a multisample integrated metabolomics strategy. Our findings offered additional insights into the molecular mechanism of the DBR-induced hepatotoxicity. We identified different metabolites from rat plasma, urine, and feces by employing gas chromatography-mass spectrometry in combination with multivariate analysis. In total, 55 metabolites distributed in 33 metabolic pathways were identified as being significantly altered in DBR-treated rats. Correlation network analysis revealed that the hub metabolites of hepatotoxicity were mainly associated with amino acid, bile acid, purine, pyrimidine, lipid, and energy metabolism. As such, DBR affected the physiological and biological functions of liver via the regulation of multiple metabolic pathways to an abnormal state. Notably, our findings also demonstrated that the multisample integrated metabolomics strategy has a great potential to identify more biomarkers and pathways in order to elucidate the mechanistic complexity of toxicity of traditional Chinese medicine.
Co-reporter:Dong-Sheng Zhao, Li-Long Jiang, Ya-Xi Fan, Lei-Chi Dong, Jiang Ma, Xin Dong, Xiao-Jun Xu, Ping Li, Hui-Jun Li
Food and Chemical Toxicology 2017 Volume 108, Part B(Volume 108, Part B) pp:
Publication Date(Web):1 October 2017
DOI:10.1016/j.fct.2017.02.030
•A targeted metabolomics of bile acids was applied to characterize the process of PMR-induced hepatotoxicity.•Hyodeoxycholic acid in serum and tauro-β-muricholic acid in urine were identified as potential biomarkers.•The expression of Bsep and Ntcp in the liver indicated that hepatotoxicity was related to the disorders of BAs metabolism.Polygoni Multiflori Radix (PMR) has been widely used as a tonic for centuries. However, hepatotoxicity cases linked to PMR have been frequently reported and appropriate biomarkers for clinical diagnosis are currently lacking. Here, an approach using UPLC-QqQ/MS-based targeted metabolomics of bile acids (BAs) complemented with biochemistry and histopathology was applied to characterize the development and recovery processes of PMR-induced hepatotoxicity in rats and to identify biomarkers. The expression of bile salt export pump (Bsep) and sodium taurocholate cotransporting polypeptide (Ntcp) were evaluated to investigate the underlying mechanism. Steatosis and inflammatory cell infiltration were observed in PMR-treated rats, which were accompanied by the elevation of serum biochemistry. The metabolic profiles of BAs were analyzed by Principal Component Analysis, hyodeoxycholic acid (HDCA) in serum and tauro-β-muricholic acid (TβMCA) in urine were identified as potential biomarkers for PMR-induced hepatotoxicity. The elevated expression of Bsep and decreased expression of Ntcp in the liver of PMRtreated rats indicated that hepatotoxicity was related to the disorders of BAs metabolism. Our study demonstrated that BAs may be used for clinical diagnosis of PMR-induced hepatotoxicity. Urine TβMCA was identified as a promising biomarker to facilitate the clinical monitoring of PMR-induced hepatotoxicity and may serve as potential therapeutic target.Download high-res image (293KB)Download full-size image
Co-reporter:Ting Pan, Tao-fang Cheng, Yu-ran Jia, Ping Li, Fei Li
Journal of Ethnopharmacology 2017 Volume 205(Volume 205) pp:
Publication Date(Web):9 June 2017
DOI:10.1016/j.jep.2017.04.020
Ethnopharmacological relevanceClematis chinensis Osbeck / Notopterygium incisum Ting ex H, T-Chang (CN) is a traditional Chinese herb couple with prominent efficacy. The herb couple has been commonly used for clinical treatment of arthralgia syndrome (“Bi Zheng” in Chinese) for centuries in China, including rheumatic arthritis, osteoarthritis and gout in modern medicine.Aim of the studyTo evaluate the anti-arthritic effect of CN herb couple in a rat model of rheumatoid arthritis (RA).Materials and methodsRats were divided randomly into six groups with eight each. Adjuvant-induced arthritis (AIA) model was established by intradermal injection of complete Freund's adjuvant (CFA). Rats were treated orally with different dosages of CN (0.7 g/kg, 2.1 g/kg, 6.3 g/kg) from day 16 till day 40. Ibuprofen (50.4 mg/kg) served as a positive control. Spontaneous activity, body weight, paw swelling, and arthritis index (AI) were monitored throughout drug treatment. Then serum levels of tumor necrosis factor α (TNF-α), interleukin-6 (IL-6), and vascular endothelial growth factor (VEGF) were determined by enzyme linked immunosorbent assay (ELISA) kits. In addition, histopathological examination and immunohistochemistry were used to assess the severity of arthritis.ResultsThree dosage of CN significantly ameliorated symptoms of RA via increasing body weight as well as reducing paw swelling (at dose of 6.3 g/kg, p<0.01) in AIA rats. An extremely significant reduction of AI (p<0.001) was also observed with treatment of CN (6.3 g/kg) compared with model group. In parallel, treatment of CN significantly down-regulated levels of TNF-α, IL-6, and VEGF both in serum (p<0.01) and in joint synovial compared with model rats. And histopathology revealed noticeable reduction in synovial hyperplasia, cartilage damage, and inflammatory infiltration by CN treatment, especially at dose of 6.3 g/kg.ConclusionsTo conclude, all results suggest that CN possesses evident anti-arthritic effects in AIA rats.Download high-res image (221KB)Download full-size image
Co-reporter:Si-Hui Nian, Hui-Jun Li, E-Hu Liu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2017 Volume 145(Volume 145) pp:
Publication Date(Web):25 October 2017
DOI:10.1016/j.jpba.2017.06.039
•The α-glucosidase inhibitory effect of AR and AFR were compared.•LC–MS/MS method was developed for determination of 7 bioactive constituents.•Chemometrics methods were employed to deal with the quantitative date.•AFR had the potential to be used as anti-diabetic medicinal resource.Comprehensive utilization of medicinal plant resources is of great significance for sustainable development of traditional Chinese medicines. In the present study, the α-glucosidase inhibitory activities of the rhizome and fibrous root of Anemarrhena Asphodeloides Bunge, were compared detailedly, and a high performance liquid chromatography coupled with electrospray ionization tandem triple quadrupole mass spectrometry (HPLC-QQQ/MS) method was developed for simultaneous quantification of seven bioactive constituents including neomangiferin, mangiferin, isomangiferin, timosaponin BII, timosaponin B, timosaponin AIII, and timosaponin N in 40 batches of samples. The results demonstrated that fibrous root extracts had more potent α-glucosidase inhibitory activity than rhizome extracts. Mangiferin and isomangiferin were abundant in fibrous root, while the analyzed saponins were rich in rhizome. Based on the chemometrics methods including principal component analysis (PCA), orthogonal partial least square discriminant analysis (OPLS-DA), and partial least square (PLS), mangiferin and isomangiferin might be mainly responsible for α-glucosidase inhibitory activity of the genus. These findings indicate that the established HPLC-QQQ/MS method was proven to be useful and efficient for quality control of Anemarrhena materials, and fibrous root had the potential to be utilized as anti-diabetic medicinal resource.Download high-res image (146KB)Download full-size image
Co-reporter:Yuanzhong Wang, Ehu Liu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2017 Volume 140(Volume 140) pp:
Publication Date(Web):5 June 2017
DOI:10.1016/j.jpba.2017.03.024
•The combination of LC–MS/MS and FT-IR was used for chemotaxonomy of Paris species.•Chemotaxonomic studies could support for infrageneric classification.•Possible biosynthetic pathways of steroid saponins in genus Paris were proposed.•22 common compounds in nine Paris species were identified by UHPLC-IT-TOFMS.•Metabolites variation was effected by both species and geographical regions.Paris species, which contain steroid saponins, have been used as herb folk medicines in Asia. In the present study, a comprehensive strategy based on liquid chromatography-tandem mass spectrometry (LC–MS/MS) and Fourier transform infrared (FT-IR) spectroscopy was firstly proposed to evaluate the chemotaxonomic relationships of nine Paris species sampled from different geographical regions in China. Principle component analysis (PCA) based on FT-IR data revealed chemical similarities in term of the nine species and geographical regions, indicating the accumulation of metabolites affected by the combination of geographical factors and species. The chemotaxonomic relationships of four species supported the morphological taxonomy and implied ancestry from P. polyphylla. After high-efficiency chromatographic separation, ions trap/time-of-flight mass spectrometry (IT-TOFMS) and triple quadrupole mass spectrometry (QQQ-MS) were used to identify unknown metabolites and simultaneously determine six key compounds (polyphyllin I, II, V, VI, VII and gracillin) in Paris species, respectively. The tentative identification of 22 steroid saponins was indicative of a common biosynthetic pathway in Paris species. Phytoecdysones, gracillin and open-chain steroid saponins were considered as key precursors. According to Pearson’s correlation analysis, an insignificant correlation was found between diosgenin-type and pennogenin-type saponins belonging to the same biosynthetic pathways in the current stage. Our results could provide a reasonable foundation for chemotaxonomy or further studies of Paris species.Download high-res image (127KB)Download full-size image
Co-reporter:Su-Ling Zeng, Li Duan, Bai-Zhong Chen, Ping Li, E-Hu Liu
Journal of Chromatography A 2017 Volume 1508(Volume 1508) pp:
Publication Date(Web):28 July 2017
DOI:10.1016/j.chroma.2017.06.015
•An efficient Visual Basic for Applications-based background subtraction program was developed in the Microsoft Office.•A novel modified mass defect filter was proposed based on different metabolic phase types.•An integrated strategy for chemicalome and metabolome characterization of polymethoxylated flavonoids in Citri Reticulatae Pericarpium was proposed.Detection of metabolites in complex biological matrixes is a great challenge because of the background noise and endogenous components. Herein, we proposed an integrated strategy that combined background subtraction program and modified mass defect filter (MMDF) data mining in a Microsoft Excel platform for chemicalome and metabolome profiling of the polymethoxylated flavonoids (PMFs) in Citri Reticulatae Pericarpium (CRP). The exogenously-sourced ions were firstly filtered out by the developed Visual Basic for Applications (VBA) program incorporated in the Microsoft Office. The novel MMDF strategy was proposed for detecting both target and untarget constituents and metabolites based on narrow, well-defined mass defect ranges. The approach was validated to be powerful, and potentially useful for the metabolite identification of both single compound and homologous compound mixture. We successfully identified 30 and 31 metabolites from rat biosamples after oral administration of nobiletin and tangeretin, respectively. A total of 56 PMFs compounds were chemically characterized and 125 metabolites were captured. This work demonstrated the feasibility of the integrated approach for reliable characterization of the constituents and metabolites in herbal medicines.
Co-reporter:Li Duan, Li-Li Dou, Ke-Yun Yu, Long Guo, Chen Bai-Zhong, Ping Li, E-Hu Liu
Food Chemistry 2017 Volume 234(Volume 234) pp:
Publication Date(Web):1 November 2017
DOI:10.1016/j.foodchem.2017.05.018
•Eight PMFs were isolated from the peel of Citrus reticulata ‘Chachi’ (CRC).•6,7,8,3′,4′-Pentamethoxyflavanone was isolated from the peel of CRC for the first time.•Eight PMFs were quantified by HPLC.•SREBPs inhibition, antiproliferative and anti-inflammatory activities of PMFs were investigated.Citrus polymethoxyflavones (PMFs) have been of increasing interest due to their extensive biological activities. In the present study, a total of eight PMFs were isolated from the peel of Citrus reticulata ‘Chachi’ (CRC). They were individually identified as 5-hydroxy-6,7,8,4′-tetramethoxyflavone (1), 5,6,7,3′,4′-pentamethoxyflavanone (2), 5-hydroxy-6,7,8,3′,4′-pentamethoxyflavone (3), 3,5,6,7,8,3′,4′- heptamethoxyflavone (4), 5,6,7,8,3′,4′-hexamethoxyflavone (5), 5,6,7,8,4′-pentamethoxyflavone (6), 6,7,8,3′,4′-pentamethoxyflavanone (7) and 5,6,7,3′,4′-pentamethoxyflavone (8) by nuclear magnetic resonance and mass spectroscopic analysis. 6,7,8,3′,4′-Pentamethoxyflavanone was isolated from the peel of CRC for the first time. The content of PMFs was firstly quantified in both the peel of CRC and PMF-rich extract by HPLC analysis. Furthermore, the biological activities of PMF compounds were investigated. 3,5,6,7,8,3′,4′-Heptamethoxyflavone demonstrated potent sterol regulatory element-binding proteins inhibition activity and 5-hydroxy-6,7,8,3′,4′-pentamethoxyflavone exhibited strong antiproliferative activity against tumor cell lines. The isolated PMF compounds could also significantly inhibit NO production and the effect varied mainly depending on the number of methoxy groups.
Co-reporter:Meng-Ning Li, Chao-Ran Li, Wen Gao, Ping Li, Hua Yang
Analytica Chimica Acta 2017 Volume 982(Volume 982) pp:
Publication Date(Web):22 August 2017
DOI:10.1016/j.aca.2017.05.030
•An HPLC-Chip-MS method was established to identify trace chemicals in complex matrix.•A database/multi-step filtering method was developed to characterize monacolin analogues.•A total of 84 monacolins including 16 new ones were identified in monascus-fermented rice product.Trace analysis of chemical analogues was always a hot topic and attracted the researchers' attentions. In this study, a database/multi-step filtering strategy was developed by using HPLC-Chip-MS system for comprehensive characterization of monacolin analogues in monascus-fermented rice product (MFRP). This strategy mainly included three steps, including chemical profile of MFRP by HPLC-Chip-MS, establishment of monacolin analogue database, multi-step filtering of monacolins based on modified mass defect filtering. All target compounds showed the symmetrical peak shapes and high MS response by using nanoflow HPLC-Chip-MS. According to the previous literature and experimental MS data, a database including 720 monacolin analogues was established. The original 522 ions in MFRP were automatically extracted by molecular feature extraction function. And then, through rectangular mass defect filtering and analogues distribution area filtering, 298 ions were further excluded. Finally, a total of 84 monacolins including 16 new compounds were unambiguously identified or tentatively characterized based on MS/MS information. In comparison with conventional mass defect filtering method, the proposed method was accurately able to exclude false positive results. The 438 false positive results were excluded in our study, while only 250 ones would be filtered out by using conventional method. This study provided a sensitive and powerful method for rapidly characterization of trace chemicals in complex matrix.Download high-res image (255KB)Download full-size image
Co-reporter:Xin-Guang Liu;Xu Lu;Ji-Xin Wang;Bin Wu;Lin Lin;Hui-Ying Wang;Ru-Zhou Guo;Hua Yang
RSC Advances (2011-Present) 2017 vol. 7(Issue 87) pp:55309-55317
Publication Date(Web):2017/12/01
DOI:10.1039/C7RA06229J
Secondary metabolites play a pivotal role in plant physiology and medicinal function. Much work has been carried out to uncover the genetic basis of plant secondary metabolite variation, but direct screening of gene variation in the whole genome is extremely time- and labor-consuming. The prediction of a candidate locus of a single nucleotide polymorphism (SNP) will save much time and resource. In this work, we combined a paired analytical metabolomics and a common garden trial to bridge the association of plant metabolism and related gene variation of Ginkgo cultivated varieties. Firstly, the leaves of 30 cultivated varieties of Ginkgo biloba L. grown in the same garden since 1990 were analyzed by UHPLC-QQQ MS/MS. Thirty-six metabolites in the flavonoid biosynthetic pathway were quantified. The biosynthetic rate of flavonoids in different cultivated varieties could reflect the related enzyme gene variation, since the environmental influence was minimized. Thus, the role of SNPs in possible varied genes was further associated with flavonoid synthesis. Results showed that after long term environment and artificial influence, different accessions of G. biloba showed a varied ability in flavonoid aglycone synthesis due to some gene polymorphism; this difference may be heritable but not obvious. Compared with previous methods, this strategy is advantageous in accuracy, low sample requirement and easier operation, providing effective information in phenotype–genotype association, and it can also be used in the heritability study of artificial breeding before large-area introduction.
Co-reporter:Jian-Guo Sun;Wei-Zhi Weng;Bo Zhang
Green Chemistry (1999-Present) 2017 vol. 19(Issue 4) pp:1128-1133
Publication Date(Web):2017/02/21
DOI:10.1039/C6GC03115C
In the presence of dimethyl sulfoxide (DMSO) as a mild oxidant and reaction medium, a simple and efficient protocol has been developed for the preparation of phosphinothioates via oxidative dehydrogenative phosphorylation of thiols with P(O)H compounds. Additionally, a DMSO-mediated oxidative phosphorylation of disulfides is also demonstrated. Notably, these transformations occur efficiently without the help of any transition metal or additive. These reactions are easy to conduct and can be scaled-up, and various phosphinothioates are readily obtained in moderate to excellent yields with excellent chemoselectivity and good functional-group tolerance.
Co-reporter:Yu-Xiang Fang;Hui-Peng Song;Jin-Xiu Liang;Hua Yang
Analytical Methods (2009-Present) 2017 vol. 9(Issue 23) pp:3422-3429
Publication Date(Web):2017/06/15
DOI:10.1039/C7AY00777A
Ultrafiltration liquid chromatography-mass spectrometry (UF-LC-MS), affinity-guided isolation, and molecular docking were integrated into one strategy for screening of enzyme inhibitors from functional foods. UF-LC-MS facilitated the rapid characterization of ligands that could bind to an enzyme; affinity-guided isolation was helpful for the efficient preparation of ligands for further bioactivity validation; and molecular docking contributed to studying ligand–protein interactions. The strategy was employed to screen pancreatic lipase inhibitors from Monascus-fermented rice (MFR). Finally, three active compounds, namely monascin, monasfluore B, and ankaflavin were discovered by UF-LC-MS and then purified by semipreparative HPLC. Their lipase-inhibitory activities were verified using an enzymatic functional assay. The Lineweaver–Burk plots showed that they inhibited lipase in a non-competitive manner, and the potential mechanisms were preliminarily explored by molecular docking. The results suggest that the strategy is effective in discovering bioactive compounds from complex systems.
Co-reporter:Bin Wu, Hong Luo, Xu Zhou, Cai-yi Cheng, Lin Lin, Bao-lin Liu, Kang Liu, Ping Li, Hua Yang
Biochimica et Biophysica Acta (BBA) - Molecular Basis of Disease 2017 Volume 1863, Issue 9(Issue 9) pp:
Publication Date(Web):1 September 2017
DOI:10.1016/j.bbadis.2017.06.011
•Succinate impaired autophagy and induced apoptosis in neurons.•Kaempferol protected mitochondrial morphological and functional integrity against succinate-mediate ischemic injury.•Kaempferol inhibited mitochondrial Drp1 recruitment and HK-II detachment through Akt activation in vitro and in vivo.Mitochondrial dysfunction is known as one of causative factors in ischemic stroke, leading to neuronal cell death. The present work was undertaken to investigate whether succinate induces neuron apoptosis by regulating mitochondrial morphology and function. In neurons, oxygen-glucose deprivation induced succinate accumulation due to the reversal of succinate dehydrogenase (SDH) activation, leading to mitochondrial fission. Kaempferol inhibited mitochondrial fission and maintained mitochondrial HK-II through activation of Akt, and thereby protected neurons from succinate-mediated ischemi injury. Knockdown of Akt2 with siRNA diminished the effect of kaempferol, indicating that kaempferol suppressed dynamin-related protein 1 (Drp1) activation and promoted HK-II mitochondrial binding dependently on Akt. Moreover, we demonstrated that kaempferol potentiated autophagy during oxygen and glucose deprivation, contributing to protecting neuron survival against succinate insult. In vivo, oral administration of kaempferol in mice attenuated the infract volume after ischemic and reperfusion (I/R) injury and reproduced the similar mitochondrial protective effect in the brain infract area. This study indicates that succinate accumulation plays a pivotal role in I/R injury-induced neuronal mitochondrial dysfunction, and suggests that modulation of Drp1 phosphorylation might be potential therapeutic strategy to protect neuron mitochondrial integrity and treat ischemic stroke.
Co-reporter:Jian-Guo Sun, Hua Yang, Ping Li, and Bo Zhang
Organic Letters 2016 Volume 18(Issue 19) pp:5114-5117
Publication Date(Web):September 13, 2016
DOI:10.1021/acs.orglett.6b02563
Visible light along with 5 mol % of rose bengal catalyzes the direct S–P(O) coupling between thiols and P(O)H compounds in the presence of air as the green oxidant. The protocol is operationally simple and amenable to gram-scale synthesis. A variety of S–P(O) coupling products can be readily prepared in moderate to excellent yields. The reaction features good functional-group tolerance, operational simplicity, and excellent practicality.
Co-reporter:Hao Zang;Jian-Guo Sun;Xin Dong;Bo Zhang
Advanced Synthesis & Catalysis 2016 Volume 358( Issue 11) pp:1746-1752
Publication Date(Web):
DOI:10.1002/adsc.201501102
Co-reporter:Li Duan, Li-Li Dou, Long Guo, Ping Li, and E-Hu Liu
ACS Sustainable Chemistry & Engineering 2016 Volume 4(Issue 4) pp:2405
Publication Date(Web):February 18, 2016
DOI:10.1021/acssuschemeng.6b00091
Deep eutectic solvents (DESs) are emerging as green and sustainable solvents for efficient extraction of bioactive compounds or drugs. This work aimed to comprehensively evaluate the potential and effectiveness of DESs for extraction of different types of natural compounds from biomass. Five Chinese herbal medicines including Berberidis Radix, Epimedii Folium, Notoginseng Radix et Rhizoma, Rhei Rhizoma et Radix, and Salviae Miltiorrhizae Radix et Rhizoma were selected to assess the efficiency of DESs on extraction of alkaloids, flavonoids, saponins, anthraquinones, and phenolic acids, respectively. Totally 43 types of choline chloride-, betaine-, and l-proline-based DESs with different polarity, viscosity, composition, and solubilization abilities were tailored to test their extraction efficiency, and the operation conditions were statistically optimized using response surface methodology to produce the most efficient process. In this work, DES solvents were first introduced to extract alkaloids and anthraquinones. The results indicated that most prepared DESs proved to be efficient solvents for extraction of alkaloids, but lower extractability for anthraquinones. The extraction capacity of DES may be correlated with their physical–chemical properties, including H-bonding interactions, polarity, viscosity, and pH. This study demonstrated that DESs were suitable green extraction solvents for selectively and efficiently extracting bioactive compounds from biomaterials.Keywords: Alkaloids; Deep eutectic solvents; Green extraction; Laevulinic acid; Natural products;
Co-reporter:Li Duan, Long Guo, Li-Li Dou, Chang-Lin Zhou, Feng-Guo Xu, Guo-Dong Zheng, Ping Li, E-Hu Liu
Food Chemistry 2016 Volume 212() pp:123-127
Publication Date(Web):1 December 2016
DOI:10.1016/j.foodchem.2016.05.141
•GC–MS based metabolomics was firstly applied for comparing the volatile oils in CRP.•15 metabolites were selected as chemical markers for discriminating CRP samples.•Antimicrobial activity of volatile oil in CP and GCP was preliminarily investigated.Citri Reticulatae Pericarpium, mainly including the pericarp of Citrus reticulata Blanco and the pericarp of Citrus reticulata ‘Chachi’, has been consumed daily as food and dietary supplement for centuries. In this study, GC–MS based metabolomics was employed to compare comprehensively the volatile constituents in Citrus reticulata Blanco and Citrus reticulata ‘Chachi’. Principal component analysis and orthogonal partial least squares discrimination analysis indicated that samples could be distinguished effectively from one another. Fifteen metabolites were finally identified for use as chemical markers in discrimination of Citri Reticulatae Pericarpium samples. The antimicrobial activity against Gram-negative and Gram-positive bacteria of the volatile oil from Citrus reticulata Blanco and Citrus reticulata ‘Chachi’ was investigated preliminarily.
Co-reporter:Bin Wu, Hui-Peng Song, Xu Zhou, Xin-Guang Liu, Wen Gao, Xin Dong, Hui-Jun Li, Ping Li, Hua Yang
Journal of Chromatography A 2016 Volume 1436() pp:91-99
Publication Date(Web):4 March 2016
DOI:10.1016/j.chroma.2016.01.062
•A strategy for screening trace enzyme inhibitors in herbal medicines was proposed.•UF-LC-QTOF MS was used to preliminarily find the α-glucosidase binders.•Docking analysis was adopted to predict the potential trace inhibitors.•Segment and exposure approach based LC-QTOF MS was applied to identify trace compounds.•11 Compounds of Ginkgo biloba extract were identified as high-quality α-glucosidase inhibitors.Screening of high potent enzyme inhibitors from herbal medicines is always lacking of efficiency due to the complexity of chemicals. The constituents responsible for the holistic effect may be trace-level chemicals, but these chemicals were covered by highly abundant compositions. To challenge this bottleneck, a strategy for screening minor bioactive compounds was proposed. It generally included four steps, (1) preliminarily find the enzyme binders by ultrafiltration; (2) optimise and predict the potential inhibitors by docking analysis; (3) selectively identify and prepare trace compounds by segment and exposure approach; (4) validate the activity and summarize the structure-activity relationship. As a case study, α-glucosidase (AGH) and Ginkgo biloba extract were used as the experimental materials. By comprehensive screening, 11 trace flavones were screened out and identified as strong AGH inhibitors. Among them, AGH inhibitory activities of syringetin and sciadopitysin were reported for the first time. Their IC50 values were 36.80 and 8.29 μM, respectively, which were lower than that of a positive control acarbose. In addition, the AGH inhibitory activities of the flavonoids could be ranked, based on a decreased order, as biflavone, flavone, flavone glycoside, flavone biglycoside. The strategy is expected to be practical and useful for screening bioactive molecules from herbal medicines, especially for the minor compounds, which will definitely accelerate the discovery of new drug candidates.
Co-reporter:Hui-Peng Song, Si-Qi Wu, Lian-Wen Qi, Fang Long, Li-Feng Jiang, Ke Liu, Hao Zeng, Zhi-Meng Xu, Ping Li, Hua Yang
Journal of Chromatography A 2016 Volume 1456() pp:176-186
Publication Date(Web):22 July 2016
DOI:10.1016/j.chroma.2016.06.009
•The pharmacophore-guided knockout/knockin chromatography was firstly established.•A novel strategy was proposed to screen drug leads and filter useless compounds.•A universal three-step method was developed to explain the function of herbs.•Functional compound combination with five criteria was explicitly defined.•The mechanisms of compounds in DQF against multiple free radicals were compared.Screening and deciphering active natural products of herbal medicines are of great importance for modern drug discovery. In this study, a novel strategy was proposed to rapidly filter ineffective compounds and target the most potential leads. The aim is to answer the key question of what components are responsible for the holistic bioactivity of an herbal product. To support the strategy, the pharmacophore-guided knockout/knockin chromatography was established for the first time. The greatest advantage of this method is that any interesting components could be automatically fished or knocked out. The method validation shows that the herbal extract was accurately reconstructed according to the experimental design. By combining with bioactivity assays, we demonstrated that “functional compound combination (FCC)”, which is the core and indispensable effective part, could be discovered from an herbal medicine and suitable as marker compounds for quality control. The applicable objects of the strategy include single herbs, herbal formulas and commercially herbal preparations. As an illustrative case study, the strategy was successfully applied to simultaneously determine active leads and the FCC in Dan-Qi formula which shows excellent free radical scavenging activity. The potential mechanisms of compounds in Dan-Qi formula reacting with three different free radicals were systematically reported for the first time. This strategy was expected to unveil the mystery of herbal medicines and inspire a natural product-based drug discovery.
Co-reporter:Wen Gao, Rui Wang, Dan Li, Ke Liu, Jun Chen, Hui-Jun Li, Xiaojun Xu, Ping Li, Hua Yang
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 117() pp:345-351
Publication Date(Web):5 January 2016
DOI:10.1016/j.jpba.2015.09.008
•A SSDMC method was developed for quality control of Lonicera flowers.•Six compounds belong to two types were assayed by using single reference standard.•The 51 batches of samples covering five Lonicera species were determined.•Principal component analysis was applied to distinguish five Lonicera species.•Iridoid glycosides were found to be a characteristic for Lonicera flowers identification.The flowers of Lonicera japonica Thunb. were extensively used to treat many diseases. As the demands for L. japonica increased, some related Lonicera plants were often confused or misused. Caffeoylquinic acids were always regarded as chemical markers in the quality control of L. japonica, but they could be found in all Lonicera species. Thus, a simple and reliable method for the evaluation of different Lonicera flowers is necessary to be established. In this work a method based on single standard to determine multi-components (SSDMC) combined with principal component analysis (PCA) for control and distinguish of Lonicera species flowers have been developed. Six components including three caffeoylquinic acids and three iridoid glycosides were assayed simultaneously using chlorogenic acid as the reference standard. The credibility and feasibility of the SSDMC method were carefully validated and the results demonstrated that there were no remarkable differences compared with external standard method. Finally, a total of fifty-one batches covering five Lonicera species were analyzed and PCA was successfully applied to distinguish the Lonicera species. This strategy simplifies the processes in the quality control of multiple-componential herbal medicine which effectively adapted for improving the quality control of those herbs belonging to closely related species.
Co-reporter:Fei Li, Tao-fang Cheng, Xin Dong, Ping Li, Hua Yang
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 117() pp:61-72
Publication Date(Web):5 January 2016
DOI:10.1016/j.jpba.2015.08.022
•HPLC–MS-based method was applied for global analysis of Shengmai injection.•Sixty-two compounds were characterized by HPLC–QTOF MS.•Twenty-one compounds were determined by HPLC-MS in selected ion monitoring mode.•The compound from Radix ophiopogonis was first detected and quantified in Shengmai injection.This study aimed to develop a specific and reliable method to comprehensively analyze the chemical constituents in Shengmai injection (SMI) using high performance liquid chromatography coupled with tandem mass spectrometry. The qualitative analysis of SMI was achieved on a Kromasil 100–5C18 column, and the results demonstrated that a total of sixty-two compounds in SMI were unambiguously assigned or tentatively identified, and further, twenty-one compounds including fourteen saponins, six lignans and one l-borneol-7-O-[β-d-apiofuranosyl (1 → 6)]-β-d-gluco-pyranoside were quantified by HPLC–MS. Furthermore, l-borneol-7-O-[β-d-apio-furanosyl (1 → 6)]-β-d-glucopyranoside, originated from Radix ophiopogonis, was identified and quantified in SMI for the first time. The method validation results indicated that the methods were simple, specific and reliable. All the investigated compounds showed good linearity (r2 ≥ 0.9992) with a relatively wide concentration range and acceptable recovery at 90.13–109.09%. Consequently, the developed methods were successfully applied to ten batches of SMI samples analysis. The proposed methods may provide a useful and comprehensive reference for the quality control of SMI, and thus to provide supporting data for the quality control and application of SMI clinically.
Co-reporter:Long Guo, Su-Ling Zeng, Yu Zhang, Ping Li, E-Hu Liu
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 117() pp:91-98
Publication Date(Web):5 January 2016
DOI:10.1016/j.jpba.2015.08.038
•An HPLC–MS method was established for qualitative and quantitative analysis of steroidal saponins in four Dioscoreae herbs.•A total of 11 saponins were characterized by HPLC–QTOF MS and 7 major saponins were quantified by HPLC–QQQ/MS.•PCA and HCA were performed to compare and discriminate the samples.•The results would be helpful in the quality control of herbal medicines from Dioscoreae species.Steroidal saponins, which exhibit multiple pharmacological effects, are the major bioactive constituents in herbal medicines from Dioscoreae species. In this study, a sensitive method based on high performance liquid chromatography-mass spectrometry (HPLC–MS) was established and validated for qualitative and quantitative analysis of steroidal saponins in four Dioscoreae herbs including Dioscoreae Nipponica Rhizome (DNR) and Dioscoreae Hypoglaucae Rhizome (DHR), Dioscoreae Spongiosae Rhizome (DSR) and Dioscoreae Rhizome (DR). A total of eleven steroidal saponins were identified by high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC–QTOF/MS). Furthermore, seven major steroidal saponins was simultaneous quantified using a high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC–QQQ/MS). The qualitative and quantitative analysis results indicated that the chemical composition of DNR, DHR and DSR samples exhibited a high level of global similarity, while the ingredients in DR varied greatly from the other three herbs. Moreover, principal component analysis (PCA) and hierarchical clustering analysis (HCA) were performed to compare and discriminate the Dioscoreae herbs based on the quantitative data. The results demonstrated the qualitative and quantitative analysis of steroidal saponins based on HPLC–MS is a feasible method for quality control of Dioscoreae herbs.
Co-reporter:Lei Liu, Bin Wu, Ke Liu, Chao-Ran Li, Xu Zhou, Ping Li, Hua Yang
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 126() pp:1-8
Publication Date(Web):15 July 2016
DOI:10.1016/j.jpba.2016.04.027
•An ionic liquid-modified MEKC method was proposed.•The addition of ILs in BGE could significantly improve the resolution of target analysts.•The micellar system was optimized by using a D-optimal design.•A 16-run experimental plan was carried out.•Eight phenolic acids were well separated in 10 min.An ionic liquid (IL)—modified micellar electrokinetic chromatography (MEKC) method was proposed for the separation and determination of eight phenolic acids. In order to increase separation efficiency and selectivity, the micelle system consisting of aqueous mixtures of ILs, Tween 20 and borate was optimized using a D-optimal design. A 16-run experimental plan was carried out. The results indicated that the addition of ILs in background electrolyte could significantly alter the electrophoretic behavior and improve the resolution of target analytes. By evaluating the electropherograms obtained, a satisfactory separation condition for all analytes was achieved in 10 min with optimized buffer composed of 0.70% (w/w) 1-butyl-3-methylimidazolium tetrafluoroborate, 8.1% (w/w) polyoxyethylene sorbitan monolaurate (Tween 20) and 10 mM sodium borate at pH 9.2. Under these conditions, all calibration curves showed good linearity (r2 > 0.9969), and accuracy (recoveries ranging from 94.71 to 106.85%). Finally, the proposed method was successfully applied to determine the phenolic acids in a Chinese medicine compound, compound danshen dripping pills.
Co-reporter:Sihui Nian;Ehu Liu;Yong Fan;Raphael N. Alolga;Huijun Li
Journal of Separation Science 2016 Volume 39( Issue 16) pp:3195-3204
Publication Date(Web):
DOI:10.1002/jssc.201600274
The green and efficient preparation of natural products from biomass is considered an important area of interest in pharmaceutical applications. In this study, we aimed to provide a practical example with a popular traditional Chinese medicine, Anemarrhenae Rhizome, and showcase the orthogonal use of column chromatography with polyamide and macroporous adsorption resins to selectively concentrate and efficiently purify four bioactive compounds: neomangiferin (NMF), mangiferin (MF), timosaponin B-II (TS B-II), and timosaponin A-III (TS A-III). First, polyamide T60–100 was employed to fractionalize the crude extracts of Anemarrhenae Rhizome. Macroporous resin HPD400 was subsequently used to purify the xanthones and steroidal saponins. Under the optimized conditions, 2.31 g of NMF, 4.10 g of MF, 12.87 g of TS B-II, and 2.78 g of TS A-III were prepared from 1 kg of crude materials, and their purities were 90.0, 92.15, 90.8, and 92.61%, respectively. The results of this study indicate that the combined column chromatography with polyamide and macroporous adsorption resins can be referenced as a green and efficient alternative for large-scale purification of bioactive ingredients from herbal raw materials.
Co-reporter:Long Guo, Li Duan, Li-Li Dou, Le-Le Liu, Hua Yang, E-Hu Liu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 129() pp:320-331
Publication Date(Web):10 September 2016
DOI:10.1016/j.jpba.2016.07.023
•The effective compounds combination (ECC) responsible for the cardioprotective efficacy of tanshinones extract (TE) was discovered.•The pharmacokinetics of single tanshinone compound, ECC and TE were studied comparatively.•A quality standardization of TE was established based on ECC using single standard for determination of multiple components method.Selection of suitable labeled constituents is vital for the quality standardization of herbal medicines (HMs). However, discovery of labeled constituents that can account for the whole efficacy of original HMs is a challenging issue. Taking tanshinones extract (TE) as an example, a strategy to establish reasonable quality control method using effective compounds combination (ECC) as labeled constituents was proposed. The strategy consists of three core steps, including chemical profiling of TE, discovery and in vivo process research of ECC, and quality standardization based on ECC. Using this strategy, a combination of four tanshinones (tanshinones IIA, cryptotanshinone, tanshinone I and dihydrotanshinone I), which was as effective as TE in cell models and in a rat model of myocardial ischemia-reperfusion, was identified as ECC in TE. Furthermore, quality standardization of TE was established based on proposed ECC using single standard for determination of multiple components (SSDMC) method. In conclusion, the presently developed ECC-based approach not only offers new insight into the understanding of the holistic effects of HMs, but also provides efficacy-associated labeled constituents for quality control of botanicals.
Co-reporter:Jimmy K. Lelu, Qi Liu, Raphael N. Alolga, Yong Fan, Wei-Lie Xiao, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 125() pp:355-359
Publication Date(Web):5 June 2016
DOI:10.1016/j.jpba.2016.04.019
•This short communication details a new automated multi-step preparative separation system, known as Sepbox.•Sepbox afforded a quick, reliable and easy separation of 11 ginsenosides from the crude extract of steamed PNG.•Inclusive in the 11 ginsenosides separated were rare ginsenosides Rh4, 20 (S) −Rg3, Rk1 and Rk5 from PNG.The root and rhizome of Panax notoginseng (PNG) are used as folk medicine. Recent studies have reported PNG to possess immunomodulatory, cardioprotective, hepatoprotective, anti-diabetic and anticancer activities among a host of other pharmacological effects. The main active constituents responsible for these pharmacological effects are saponins. It has also been proven that the chemical constituents of steamed PNG differs from the raw form. Traditional methods of separating individual components in crude extracts are usually tedious, almost irreproducible and time-consuming. In this study, an automated multi-step preparative separation system, known as Sepbox afforded a quick, reproducible and fast separation of saponins from PNG. With Sepbox, a total of 11 saponins of high purity were obtained in a short period of time. The separated compounds were identified as notoginsenosides R1, T5, ginsenosides Rb1, Rg1, Rg2, Rh1, Rh4, Rd, 20 (S) −Rg3 and a mixture of ginsenosides Rk1 and Rg5.
Co-reporter:Shao-Teng Wang, Hua Yang, Wen Gao, Hui-Jun Li, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 119() pp:91-98
Publication Date(Web):5 February 2016
DOI:10.1016/j.jpba.2015.11.033
•Polyphenols were selectively enriched by a weak anion-exchange SPE.•MS/MS fragmentation rules of polyphenols were outlined.•Twenty six trace members were identified for the first time in this herb.The analysis of trace constituents in herbal medicines has always been a challenge due to complex matrices and structural diversities. In this work, a pH-sensitive solid phase extraction (SPE) procedure capable of enriching trace polyphenols in Bistort Rhizoma (BR) was proposed and preliminary chemical characterization was accomplished by high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (HPLC-QTOF MS). A weak anion-exchange SPE column packed with divinylbenzene/vinylpyrrolidone bonding quaternary amine group was employed for anionic extraction, and the target fraction was obtained by eluting with acidic methanol (apparent pH 1.9). On the other hand, the MS/MS fragmentation rules of four reference polyphenols in negative ion mode were outlined. Using these rules, a total of 31 polyphenols including 20 benzoyl derivatives and 11 caffeoyl derivatives were screened out from BR extract, of which 26 trace members were found for the first time in this herb. Those findings demonstrated that the anion-exchange SPE could enhance the detection capability and selectivity for plant polyphenols in the LC–MS analysis and the strategy for deducing structures could be applied for analysis of polyphenols in BR and other herbal medicines.
Co-reporter:Tao-fang Cheng;Yu-ran Jia;Zheng Zuo;Xin Dong;Ping Zhou;Fei Li
Journal of Separation Science 2016 Volume 39( Issue 7) pp:1223-1231
Publication Date(Web):
DOI:10.1002/jssc.201501259
This study was designed to develop a simple, specific and reliable method to overall analyze the chemical constituents in clematidis radix et rhizome/notopterygii rhizome et radix herb couple using high-performance liquid chromatography coupled with tandem mass spectrometry and multiple chemometric analysis. First, the separation and qualitative analysis of herb couple was achieved on an Agilent Zorbax Eclipse Plus C18 column (250 mm × 4.6 mm, 5 μm), and 69 compounds were unambiguously or tentatively identified. Moreover, in quantitative analysis, eight ingredients including six coumarins and two triterpenoid sapogenins were quantified by high-performance liquid chromatography coupled with tandem mass spectrometry. In terms of good linearity (r2 ≥ 0.9995) with a relatively wide concentration range, recovery (85.40–102.50%) and repeatability (0.99–4.45%), the validation results suggested the proposed method was reliable, and successfully used to analyze ten batches of herb couple samples. Then, hierarchical cluster analysis and principal component analysis were used to classify samples and search significant ingredients. The results showed that ten batches of herb couple samples were classified into three groups, and six compounds were found for its better quality control.
Co-reporter:Wen Gao, Xin Dong, Rui Wang, Xin-Guang Liu, Ping Li and Hua Yang
RSC Advances 2016 vol. 6(Issue 66) pp:61418-61422
Publication Date(Web):16 Jun 2016
DOI:10.1039/C6RA10975F
Due to the low viscosity of carbon dioxide, supercritical-fluid chromatography (SFC) allows higher flow rates with lower back pressure. Generally, analytical SFC could provide much faster and more efficient separation compared to conventional chromatographic methods. However, the non-polar characteristic of supercritical carbon dioxide (scCO2) has limited the applicability of SFC in the separation of hydrophobic compounds. Ionic liquids (ILs), with attractive features including a wide range of solubility, miscibility, and low vapor pressure, may offer a possibility to improve SFC separation. In this work, an analytical SFC method using IL as mobile phase modifier was developed for the separation of six flavonoid aglycones, and 1-butyl-3-methyl imidazolium-based ILs were tested and compared as mobile phase additives. The results demonstrated the addition of ILs into SFC mobile phase could significantly improve the resolution of flavonoid aglycones, and the selectivity changed probably based on the hydrogen-bonding interaction.
Co-reporter:Shao-Teng Wang, Wen Gao, Ya-Xi Fan, Xin-Guang Liu, Ke Liu, Yuan Du, Ling-Li Wang, Hui-Jun Li, Ping Li and Hua Yang
RSC Advances 2016 vol. 6(Issue 33) pp:27320-27328
Publication Date(Web):02 Mar 2016
DOI:10.1039/C6RA00687F
Bistort Rhizoma (BR) is consumed as a traditional medicinal herb in China, but there reports on its bioactive components are scarce. In the present study, a fingerprinting analysis was built in 35 batches of BR from various sources, and five major compounds, the content of total phenols (TP) as well as total extractable tannins (TET) were measured. The antioxidant properties were examined by three methods, and then the correlation coefficients between antioxidant activity and content of BR properties were evaluated. The results showed that the content of TP was significantly correlated with the herbal antioxidant activity (R2 = 0.5501 in the DPPH assay; R2 = 0.6387 in the ABTS assay and R2 = 0.6722 in the FRAP assay) but the five major components are not, TET also partly influenced the herbal efficiency. In conclusion, the properties responsible for the antioxidant activity of BR are phenols, and the active constituents may be some trace phenols in BR, which need further study to unravel the truly active compounds.
Co-reporter:Xin Dong, Rui Wang, Xu Zhou, Ping Li, Hua Yang
Journal of Chromatography B 2016 Volume 1026() pp:15-26
Publication Date(Web):15 July 2016
DOI:10.1016/j.jchromb.2015.11.048
Traditional Chinese medicines (TCMs) are gaining more and more attentions all over the world. The focus of TCMs researches were gradually shifted from chemical research to the combination study of chemical and life sciences. However, obtaining precise information of TCMs process in vivo or in vitro is still a bottleneck in bioanalysis of TCMs for their chemical composition complexity. This paper reviewed the recent analytical methods especially mass spectrometry technology in the bioanalysis of TCMs, and data processing techniques in the qualitative and quantitative analyses of metabolite of TCMs. Additionally, the difficulties encountered in the analyzing biological samples in TCMs and the solutions to these problems have been mentioned.
Co-reporter:Chang-Jiang-Sheng Lai, Ting Tan, Su-Ling Zeng, Lin-Ru Xu, Lian-Wen Qi, E-Hu Liu and Ping Li
Green Chemistry 2015 vol. 17(Issue 4) pp:2580-2586
Publication Date(Web):19 Feb 2015
DOI:10.1039/C5GC00091B
Comprehensive quantification of a specific class of natural products using liquid chromatography tandem mass spectrometry with greener approaches is of significant importance. In this study, a green protocol was proposed for determining analogues with a small number of reference standards. The protocol was conducted by combining the chromatography-based fraction collection, the calibration curve matrix materials and the identical enzymatic hydrolysate. This strategy can accurately calibrate the concentration ratios and successfully calculate the relative response factors (F) of the specific protopanoxadiol-type ginsenosides. The recoveries of all fractions were monitored by the modified non-standard recovery evaluation strategy rather than the commonly used method with total reference standards. Coupled with the calculated F, the quantitative analysis of multi-components with a single marker was successfully applied to simultaneously determine the analogues of interest in Sanqi (Panax notoginseng) extract. The use of snailase for quantification of ginsenosides especially in application to the commercially unavailable ginsenoside Rd isomer was reported for the first time. By consuming less solvent and fewer reference standards, the developed green protocol facilitates its application to other analogues in herbal and food samples by utilizing suitable enzymes or derivative techniques.
Co-reporter:Hui-Peng Song, Jun Chen, Jia-Ying Hong, Haiping Hao, Lian-Wen Qi, Jun Lu, Yu Fu, Bin Wu, Hua Yang and Ping Li
Chemical Communications 2015 vol. 51(Issue 8) pp:1494-1497
Publication Date(Web):02 Dec 2014
DOI:10.1039/C4CC08728C
A novel strategy of ultrafiltration LC-MS and in silico molecular docking was proposed to discover high-quality enzyme inhibitors from herbal medicines. Using this strategy, two compounds were predicted and finally demonstrated as potent xanthine oxidase inhibitors, whose in vitro IC50 values were lower than that of a positive control allopurinol.
Co-reporter:Ru-Zhou Guo, Xin-Guang Liu, Wen Gao, Xin Dong, Salvatore Fanali, Ping Li, Hua Yang
Journal of Chromatography A 2015 Volume 1422() pp:147-154
Publication Date(Web):27 November 2015
DOI:10.1016/j.chroma.2015.10.008
•Separation of Ginkgo biloba extract was conducted by comprehensive LC × LC.•Screening antioxidant by comprehensive LC × LC–pre-column DPPH was proposed.•Sixteen compounds were firstly reported of free radical scavenging activity.•Performance of comprehensive LC × LC was compared with conventional HPLC.Recently, screening of bioactive compounds by on-line ultra-high performance liquid chromatography (UHPLC) draws increasing attentions for the advantages of rapidity and intuition. Nevertheless, most on-line methods were limited to the shortcoming like low resolution and peak capacity, which could interfere the active ingredient identification. Comprehensive two-dimensional UHPLC (LC × LC) has revealed to be a powerful tool to separate complex mixtures. Herein, a strategy based on LC × LC analysis coupled with pre-column 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay was proposed to screen the antioxidants from the extract of Ginkgo biloba (EGB). A total of 61 compounds were identified in EGB, and 25 of them showed appreciable radical scavenging capacity. This work may offer pharmacodynamics base for further research about EGB, also the strategy is likelihood to be applied in screening antioxidant in other herbal medicine.
Co-reporter:Li-Chao Wang, Wen-Song Zhang, Qun Liu, Jia Li, Raphael N. Alolga, Kang Liu, Bao-Lin Liu, Ping Li, Lian-Wen Qi
Journal of Functional Foods 2015 Volume 16() pp:20-27
Publication Date(Web):June 2015
DOI:10.1016/j.jff.2015.04.018
•RGSE attenuated acute myocardial infarction-induced heart injury.•RGSE inhibited oxidative stress and apoptosis during AMI in vivo and in vitro.•TXNIP-NLRP3-IL-1β signaling, for the first time, was revealed to be activated in AMI, promoting cell death in the end.•RGSE exerted cardioprotection by suppression of TXNIP/NLRP3 activation during ER stress.•Ginsenosides Rh4, Rg5, Rk3, and Rk1 were the most abundant, and notoginsenoside T5/isomer was first identified in steamed notoginseng.Panax notoginseng possesses therapeutic potential for cardiovascular disorders. Endoplasmic reticulum (ER) stress plays a critical role in cardiovascular disease. Here we show that a rare ginsenoside-standardized extract (RGSE) from notoginseng exerts cardioprotection via attenuating ER stress-associated apoptosis. A total of 18 ginsenosides with a concentration of 816.75 mg/g were characterized and quantified in RGSE. Ginsenoside Rh4, Rg5, Rk3, and Rk1 were the most abundant. RGSE at 50 and 100 mg/kg significantly attenuated myocardial infarction and improved cardiac function in acute myocardial infarction (AMI). RGSE also prevented myocardial ischemia-induced oxidative stress and apoptosis in vivo and in vitro. In addition, RGSE decreased the overexpression of GRP78, GRP94 and CHOP, and inhibited the phosphorylation of PERK and IRE1α in ER stress. Further investigation revealed that RGSE effectually inactivated TXNIP/NLRP3 signaling pathway in AMI hearts. These observations suggest a novel role for notoginseng in regulating AMI.
Co-reporter:Peipei Wang, Lei Zhang, Jian Yao, Yikang Shi, Ping Li, Kan Ding
Carbohydrate Polymers 2015 Volume 121() pp:328-335
Publication Date(Web):5 May 2015
DOI:10.1016/j.carbpol.2014.11.073
•A novel arabinogalactan RN1 was purified from flowers of Panax notoginseng.•Backbone of RN1 is 1,6-linked Galp, attached by a branch with 1,3-linked Galp at C-3.•RN1 could inhibit angiogenesis by disrupting BMP2/ Smad1/5/8/Id1 signalingAngiogenesis plays an essential role in tumor development. Blocking angiogenesis in tumor has become a promising tactic in limiting cancer progression. Here, an arabinogalactan polysaccharide, RN1 was isolated from flowers of Panax notoginseng. Its structure was determined to possess a backbone of 1,6-linked Galp branched at C3 by side 1,3-linked Galp, with branches attached at position O-3 of it. The branches mainly contained 1,5-linked, 1,3,5-linked, terminal Arabinose and terminal Galactose. RN1 could inhibit microvessel formation in the BxPC-3 pancreatic cancer cell xenograft tumor in nude mice. The antiangiogenesis assay showed that RN1 could reduce the migratory activity of endothelial cells and their ability of tube formation on matrigel, but no effect on endothelial cells growth. Further studies revealed that RN1 could inhibit BMP2/Smad1/5/8/Id1 signaling. All those data indicated the RN1 had an antiangiogenic effect via BMP2 signaling and could be a potential novel inhibitor of angiogenesis.
Co-reporter:Pan Zhao, Li Duan, Long Guo, Li-Li Dou, Xin Dong, Ping Zhou, Ping Li, E-Hu Liu
Food Chemistry 2015 Volume 173() pp:54-60
Publication Date(Web):15 April 2015
DOI:10.1016/j.foodchem.2014.10.010
•TLC and HPLC–QTOF/MS techniques were employed to compare the chemical profiles of CWT and CML.•A HPLC–DAD method was developed for quantification of eight major compounds in CWT and CML.•The antioxidant activity of CWT and CML were compared by DPPH, FRAP and ABTS assay.•The results indicated that there were significant chemical differences between CWT and CML, and CWT had higher antioxidant activity than CML.Citri Fructus (CF), the mature fruit of Citrus wilsonii Tanaka (CWT) or Citrus medica L. (CML), is an important citrus by-product with health promoting and nutritive properties. The present study compares the chemical and biological differences of CWT and CML. Thin layer chromatography and high performance liquid chromatography, coupled with quadrupole time-of-flight tandem mass spectrometry techniques, were employed to compare the chemical profiles of CWT and CML. A total of 25 compounds were identified and the results indicated that there were significant differences in chemical composition between the two CF species. The quantitative results obtained by HPLC coupled with diode array detector method demonstrated that naringin was present in the highest amounts in CWT, whilst nomilin was the most dominant constituent in CML. It was also found that CWT had significantly higher free radical-scavenging activity than CML.
Co-reporter:Lei Zhang;Kai Song;Ling Zhou;Zhishen Xie;Ping Zhou;Yiming Zhao;Yue Han;Xiaojun Xu
Journal of Cellular Biochemistry 2015 Volume 116( Issue 6) pp:1101-1112
Publication Date(Web):
DOI:10.1002/jcb.25066
ABSTRACT
Heparan sulfate (HS) are complex polysaccharides that reside on the plasma membrane of almost all mammalian cells, and play an important role in physiological and pathological conditions. Heparan sulfate D-glucosamine 3-O-sulfotransferase 3B1 (HS3ST3B1) participates in the last biosynthetic steps of HS and transfers sulfate to the 3-O-position of glucosamine residues to yield mature sugar chains. To date very few biological processes or proteins have been described that are modulated by HS3ST3B1. In this study, we observed that HS3ST3B1 positively contributed to acute myeloid leukemia (AML) progression in vitro and in vivo, and these activities were associated with an induction of the proangiogenic factor VEGF expression and shedding. Moreover, the effects of HS3ST3B1 on VEGF release can be attenuated after treatment of heparanase inhibitor suramin, which prevented VEGF secretion and subsequently blocked VEGF-induced activation of ERK and AKT, suggesting that 3-O-sulfation of HS by HS3ST3B1 facilitated VEGF shedding; the effects of HS3ST3B1 on activation of ERK and AKT can also be blocked by VEGFR inhibitor axitinib, suggestive of a relationship between 3-O-sulfation of HS and VEGF-activated signaling pathways. Taken together, our findings support that VEGF is an important functional target of HS3ST3B1 and provide a new mechanism of HS3ST3B1 in AML. J. Cell. Biochem. 116: 1101–1112, 2015. © 2014 Wiley Periodicals, Inc.
Co-reporter:Long Guo, Li Duan, Xin Dong, Li-Li Dou, Ping Zhou, Ping Li, E.-Hu Liu
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 107() pp:473-479
Publication Date(Web):25 March 2015
DOI:10.1016/j.jpba.2015.01.043
•The metabolic profile of miltirone was characterized in detail.•HPLC/Q-TOF-MS coupled with MDF method was developed for metabolites identification.•Total 15 metabolites and 6 metabolic pathways were found in vivo.Miltirone is one of the bioactive diterpene quinones isolated from Salvia miltiorrhiza Bunge. This compound has been found to possess significant anticancer, antibacterial, antioxidant, and anti-inflammatory activities. However, the metabolic fate of miltirone remains unknown. In order to explore whether miltirone is extensively metabolized, we investigated the metabolites of miltirone in plasma, bile, urine, and feces samples following oral and intravenous administration to the rats. By using high performance liquid chromatography/quadrupole time-of-flight mass spectrometry (HPLC/Q-TOF-MS) coupled with mass detect filter (MDF) method, a total of 15 metabolites were identified from the biosamples. Both phase I and phase II metabolites were observed in the metabolic profile and the metabolic pathways involved in reduction, oxidation, monohydroxylation, dihydroxylation, glucuronidation and sulfation. The results indicated that hepatocyte metabolism was the major route of clearance for the parent compound. The present study provided valuable information for better understanding of the efficacy and safety of miltirone.
Co-reporter:Chang-jiang-sheng Lai, Ting Tan, Su-ling Zeng, Xin Dong, E-Hu Liu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 102() pp:150-156
Publication Date(Web):5 January 2015
DOI:10.1016/j.jpba.2014.09.004
•The MPC–HPLC–ESI-MS was developed and applied for relative quantification of Sanqi saponins.•It was applied for coeluent interference evaluation.•The normalized relative quantification data of ginsenosides were obtained.Relative quantification of multi-components in complex mixture is significantly affected by the ionization variance caused by mobile phase composition in high-performance liquid chromatography with electrospray ionization mass spectrometry (HPLC–ESI-MS) analyses. The normalization methods for eliminating the variance are still less investigated. Herein, the mobile-phase compensation (MPC) method was applied to overcome the above problem. The developed method was firstly used for convenient evaluation of the coeluent interference and subsequently applied for relative quantification of the identified multi-components in Panax notoginseng (Sanqi) samples. The good linearity, precision and low limit of quantification of targeted analytes confirmed that the MPC–HPLC–ESI-MS method in gradient elution could achieve the isocratic test results compared with classical HPLC–ESI-MS. The established method was used for relative quantification of the minor Sanqi saponins by their detected peak areas divided by that of ginsenoside Rd. The results demonstrated the potential of the newly developed method for obtaining the normalized data shared in different laboratories.The MPC–HPLC–ESI-MS method and its application.
Co-reporter:Yu Zhang, Long Guo, Li Duan, Xin Dong, Ping Zhou, E.-Hu Liu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 102() pp:110-118
Publication Date(Web):5 January 2015
DOI:10.1016/j.jpba.2014.09.006
•A HPLC–MS/MS was established for determination of 16 phenolic compounds in Spatholobi Caulis.•PCA was performed to compare and discriminate the samples.•The results would be helpful in evaluation of the quality of Spatholobi Caulis.Spatholobi Caulis, the vine stem of Spatholobus suberectus Dunn, has been widely used in traditional Chinese and folk medicines for treatment of irregular menstruation, blood deficiency and rheumatalgia in clinic. In this study, an accurate and reliable high performance liquid chromatography/electrospray ionization triple quadrupole mass spectrometry (HPLC–MS/MS) method was established for simultaneous determination of 16 phenolic bioactive constituents, including five flavanols, seven isoflavonoids, three flavanones and one chalcone in Spatholobi Caulis. The method validation results exhibited that the developed method had desirable specificity, linearity, precision and accuracy. The quantitative analysis results showed that flavanols were the abundant constituents in Spatholobi Caulis. Moreover, principal component analysis (PCA) was performed to assess the quality variation of samples collected from different regions in China. The PCA results indicated the quantitative analysis based on HPLC–MS/MS is a feasible method for quality assessment and control of Spatholobi Caulis.
Co-reporter:Jun Lu, Hui-Peng Song, Ping Li, Ping Zhou, Xin Dong, Jun Chen
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 109() pp:85-90
Publication Date(Web):10 May 2015
DOI:10.1016/j.jpba.2015.02.020
•19 compounds were screened out as direct thrombin inhibitors.•Three compounds were discovered to inhibit thrombin for the first time.•Docking results gave a reasonable explanation for thrombin–ligand interaction.•The global screening method was universal for other enzymes.Thrombin plays a significant role in thromboembolic disease. In this work, a peak fractionation approach combined with an activity assay method was used to screen direct thrombin inhibitors from Radix Salviae Miltiorrhizae (RSM), a famous herbal remedy for the treatment of cardiovascular diseases in China. A total of 91 fractions were collected from the RSM extract, and 19 fractions out of them showed thrombin inhibitory effects with dose–effect relationship. Among them, three compounds were unambiguously identified as 15, 16-dihydrotanshinone I, cryptotanshinone and tanshinone IIA with IC50 values of 29.39, 81.11 and 66.60 μM, respectively. The three compounds were reported with direct thrombin inhibition activities for the first time and their ligand–thrombin interactions were explored by a molecular docking research. These results may contribute to explain the medical benefit of RSM for the prevention and treatment of cardiovascular diseases.
Co-reporter:Chang-Jiang-Sheng Lai, Ting Tan, Su-Ling Zeng, Lian-Wen Qi, Xin-Guang Liu, Xin Dong, Ping Li, E-Hu Liu
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 109() pp:184-191
Publication Date(Web):10 May 2015
DOI:10.1016/j.jpba.2015.02.028
•The segment and exposure strategy increased the MS detection efficiency.•The five-point screening approach was developed for screening ginsenosides.•The visual isotopic ion technique was used for verifying the screened ginsenosides.•Two hundred thirty four saponins including 67 new ones were identified from Panax notoginseng.The aim of this study was to develop a convenient method without pretreatments for nontarget discovery of interested compounds. The segment and exposure strategy, coupled with two mass spectrometer data acquisition methods was firstly proposed for screening the saponins in extract of Panax notoginseng (Sanqi) via high-performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry (HPLC-QTOF/MS). By gradually removing certain major or moderate interference compounds, the developed segment and exposure strategy could significantly improve the detection efficiency for trace compounds. Moreover, the newly developed five-point screening approach based on a modified mass defect filter strategy and the visual isotopic ion technique was verified to be efficient and reliable in picking out the interested precursor ions. In total, 234 ginsenosides including 67 potential new ones were characterized or tentatively identified from the extract of Sanqi. Particularly, some unusual compounds containing the branched glycosyl group or new substituted acyl groups were firstly reported. The proposed integrated strategy held a strong promise for analyses of the complex mixtures.Graphical abstract
Co-reporter:Xin-Guang Liu, Si-Qi Wu, Ping Li, Hua Yang
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 113() pp:212-225
Publication Date(Web):10 September 2015
DOI:10.1016/j.jpba.2015.03.006
•The ginkgo flavonoid related articles (from 2009 to 2014) were reviewed.•Chemical composition and routine analysis of ginkgo flavonoid were summarized.•Evaluation criterion of ginkgo flavonoid purification was discussed.•Direct and indirect quantitative methods of ginkgo flavonoid were compared.Flavonoids are the main active constituents in Ginkgo biloba L., which have been suggested to have broad-spectrum free-radical scavenging activities. This review summarizes the recent advances in the chemical analysis of the flavonoids in G. biloba and its finished products (from 2009 to 2014), including chemical composition, sample preparation, separation, detection and different quality criteria. More than 70 kinds of flavonoids have been identified in this plant. In this review, various analytical approaches as well as their chromatographic conditions have been described, and their advantages/disadvantages are also compared. Quantitative analyses of Ginkgo flavonoids applied by most pharmacopeias start with an acidic hydrolysis followed by determination of the resulting aglycones using HPLC. But increasing direct assay of individual flavonol glycosides found that many adulterated products were still qualified by the present tests. To obtain an authentic and applicable analytical approach for quality evaluation of Ginkgo and its finished products, related suggestions and opinions in the recent publications are mainly discussed in this review. This discussion on chemical analyses of Ginkgo flavonoids will also be found as a significant guide for widely varied natural flavonoids.
Co-reporter:Hua Yang, Wen Gao, Lei Liu, Ke Liu, E-Hu Liu, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 115() pp:10-19
Publication Date(Web):10 November 2015
DOI:10.1016/j.jpba.2015.06.021
•A strategy was proposed to discover chemical markers for herbal quality control.•Chromatographic profile of 3 processed aconite herbs were performed.•A compound/species-based classifer was established by probabilistic neural network.•Contribution of each compound toward species classification was calculated.•Characteristic markers were obtained from 26-batch samples covering 3 aconite herbs.Most Aconitum species, also known as aconite, are extremely poisonous, so it must be identified carefully. Differentiation of Aconitum species is challenging because of their similar appearance and chemical components. In this study, a universal strategy to discover chemical markers was developed for effective authentication of three commonly used aconite roots. The major procedures include: (1) chemical profiling and structural assignment of herbs by liquid chromatography with mass spectrometry (LC-MS), (2) quantification of major components by LC-MS, (3) probabilistic neural network (PNN) model to calculate contributions of components toward species classification, (4) discovery of minimized number of chemical markers for quality control. The MS fragmentation pathways of diester-, monoester-, and alkyloyamine-diterpenoid alkaloids were compared. Using these rules, 42 aconite alkaloids were identified in aconite roots. Subsequently, 11 characteristic compounds were quantified. A component–species modeling by PNN was then established combining the 11 analytes and 26-batch samples from three aconite species. The contribution of each analyte to species classification was calculated. Selection of fuziline, benzoylhypaconine, and talatizamine, or a combination of more compounds based on a contribution order, can be used for successful categorization of the three aconite species. Collectively, the proposed strategy is beneficial to selection of rational chemical markers for the species classification and quality control of herbal medicines.
Co-reporter:Meng-Ning Li, Xin Dong, Wen Gao, Xin-Guang Liu, Rui Wang, Ping Li, Hua Yang
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 114() pp:376-389
Publication Date(Web):10 October 2015
DOI:10.1016/j.jpba.2015.05.030
•UHPLC-QTOF MS method was developed for global identification of Qi-Fu-Yin.•A total of 156 compounds were characterized by UHPLC-QTOF MS.•UHPLC-QQQ MS with a fast polarity switching mode was applied to determine 26 major compounds.•The performance of UHPLC-QQQ MS was compared with the conventional method.Qi-Fu-Yin (QFY), a classical traditional Chinese medicine formula, is proven to have significant neuroprotective effects by modern pharmacological studies. However, the chemical constituents of QFY have not been fully explored. In this study, an ultra-high performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-QTOF MS) was developed for comprehensive analysis of chemical constituents in QFY. By using characteristic ions and fragmentation rules, a reliable identification of 156 compounds was described here, including 69 triterpene saponins, 23 oligosaccharide esters, 22 flavanoids, 9 alkaloids, 9 phenolic acids, 8 phthalides, 7 phenylethanoid glycosides, 3 xanthones, 3 sesquiterpene lactones, 2 ionones and 1 iridoid glycoside. Twenty-six major compounds were then determined in a single run by UHPLC coupled with triple quadrupole tandem mass spectrometry (QQQ MS) with fast positive/negative polarity switching. It allows for the acquisition of MS data in both ionization modes from a single run. The proposed method was then validated in terms of linearity, accuracy, precision and recovery. The overall recoveries for 26 analytes ranged from 91.35% to 109.58%, with RSDs ranging from 0.82% to 4.83%. In addition, the content of 26 analytes in QFY prepared by five batches of herbal materials was also analyzed. These results demonstrated that our present method was effective and reliable for comprehensive quality evaluation of QFY. Meanwhile, the study might provide the chemical evidence for revealing the material basis of its therapeutic effects.
Co-reporter:Ting-Wen Yang, Chao Zhao, Yong Fan, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 114() pp:280-287
Publication Date(Web):10 October 2015
DOI:10.1016/j.jpba.2015.05.028
•The effect of UV wavelength and standard solution concentrations on RRF was studied.•A peak area ratio-wavelength curve was given to help choose UV wavelength.•Line regression equation and one single point methods to calculate RRF were compared.•Suggestions on concentrations were given when using one single point method.Single standard to determine multi-components (SSDMC) is a practical pattern for quality evaluation of herbal medicines (HMs). However, it remains challenging because of potential inconsistency of relative response factors (RRF) on different instruments. In this work, the effects of two key roles, i.e., ultraviolet (UV) wavelength and standard solution concentrations, on reproducibility of RRF were investigated. The effect of UV wavelength on reproducibility of RRF was studied by plotting the relationship of the peak area ratios (internal standard vs analyte) to wavelengths. The preferable wavelength should be set at the flat parts of the curve. Optimized 300 nm produced a 0.38% RSD for emodin/emodin-8-O-β-D-glucopyranoside on five instruments, much lower than 2.80% obtained from the maximum wavelength at 290 nm. Next, the effects of standard solution concentrations of emodin on its response factor (RF) were investigated. For one single point method, low concentration less than 49b/k resulted in significant variations in RF. For emodin, when the concentration is higher than 7.00 μg mL−1, a low standard deviation (SD) value at 0.13 was obtained, while lower than 7.00 μg mL−1, a high SD at 3.71 was obtained. The developed SSDMC method was then applied to determination of target components in 10 Polygonum cuspidatum samples and showed comparable accuracy to conventional calibration methods with deviation less than 1%.
Co-reporter:Jin-Yi Wan, Yong Fan, Qing-Tao Yu, Ya-Zhong Ge, Chen-Pu Yan, Raphael N. Alolga, Ping Li, Zhong-Hua Ma, Lian-Wen Qi
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 107() pp:89-97
Publication Date(Web):25 March 2015
DOI:10.1016/j.jpba.2014.11.014
•Malonyl-ginsenosides in ginseng products were characterized by LC–Q-TOF/MS.•Twelve malonyl ginsenosides were rapidly screened by the characterized ions.•Contents of 17 amino acids and polysaccharides in ginseng products were determined.•Arginine, glutamic acid, and aspartic acid were abundant in ginseng products.•Optimal time for the accumulation of ingredients was suggested to be 5–6 years.Many analytical methods have been developed to characterize ginsenosides in ginseng. Relatively less attention has been paid to the malonyl ginsenosides, amino acids and polysaccharides in various processing ginsengs. In this study, malonyl ginsenosides were characterized by LC–Q-TOF/MS. In positive mode, the most abundant ions at m/z 425.38 were observed corresponding to the protopanoxadiol-type ginsenosides. A rich diagnostic ion at 835.48 was shown representing the malonyl ginsenosides with at least two glucosides. Twelve malonyl ginsenosides were rapidly screened using 835.48–835.49 to restructure ion chromatograms. In negative mode, besides the high deprotonated ion, a neutral loss of 44 Da (CO2) was found. High-energy collision-induced dissociation at 50 V produced the most abundant product ion [M−H-malonyl]− by a neutral loss of 86 Da. Determination of 17 common amino acids was performed on an automatic amino acid analyzer. Arginine, glutamic acid, and aspartic acid were abundant. The contents of amino acids were 9.1% in fresh ginseng and 3.1% in black ginseng. Phenol-sulfuric acid method was applied to analysis of polysaccharides. The contents of polysaccharides were 29.1% in fresh ginseng and 11.1% in black ginseng. The optimal growth age for the accumulation of constituents was supposed to be 5–6 years. In conclusion, the contents of malonyl ginsenosides, amino acids, and polysaccharides, based on decreasing order, ranked as follows: fresh ginseng > frozen ginseng > white ginseng > stoved ginseng > red ginseng > black ginseng. Processing should be paid more attention for the quality control of ginseng products.
Co-reporter:Feng-Zhen Wang, Zhi-Shen Xie, Lei Xing, Bing-Feng Zhang, Jia-Liang Zhang, Peng-Fei Cui, Jian-Bin Qiao, Kun Shi, Chong-Su Cho, Myung-Haing Cho, Xiaojun Xu, Ping Li, Hu-Lin Jiang
Biomaterials 2015 73() pp: 149-159
Publication Date(Web):
DOI:10.1016/j.biomaterials.2015.09.013
Co-reporter:Zhi-Ying Fan, Xin-Guang Liu, Ru-Zhou Guo, Xin Dong, Wen Gao, Ping Li, Hua Yang
Journal of Chromatography B 2015 Volume 988() pp:1-7
Publication Date(Web):15 April 2015
DOI:10.1016/j.jchromb.2015.02.015
•A simple and sensitive UHPLC-MS/MS method was developed to determinate ginkgolide K in rat plasma and tissues.•The method was fully validated and successfully applied to pharmacokinetic study of ginkgolide K.•Systemic exposure to ginkgolide K in rats was investigated.•Ginkgolide K rapidly eliminated in 4 h and accumulated mostly in liver and kidney.Ginkgolide K (GK), a derivative compound of ginkgolide B, has been recently isolated from the leaves of Ginkgo biloba. It is a powerful natural platelet activate factor (PAF) antagonist, and also has obvious protect effects for cerebral ischemia. However, no reports have been described for the pharmacokinetic study of GK. In this study, a simple, sensitive and reliable ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method has been developed for the determination of GK in rat plasma and tissues. Biological samples were pretreated by an efficient liquid–liquid extraction with ethyl acetate. The chromatographic separation was achieved on an Agilent ZORBAX SB-Aq column (4.6 mm × 50 mm, 1.8 μm) with a mobile phase of 0.5% aqueous formic acid (A)–menthol (B). Quantitation was carried out on a triple quadruple mass spectrometry using positive electrospray ionization in multiple reaction monitoring mode. Diazepam was used as internal standard (IS). The ion transitions monitored were set at m/z 407.10 → 389.20 and m/z 285.08 → 193.10 for GK and IS, respectively. The developed method was fully validated and successfully applied to the pharmacokinetics and tissue distribution study of GK after intravenous administration. The current results have indicated that pharmacokinetic parameters of GK vary in a dose-dependent manner with rapid elimination in 4 h. The major distribution tissues of GK in rats were liver and kidney. This study would provide critical information to promote the future study of GK.
Co-reporter:Xin-Guang Liu, Lian-Wen Qi, Zhi-Ying Fan, Xin Dong, Ru-Zhou Guo, Feng-Chang Lou, Salvatore Fanali, Ping Li, Hua Yang
Journal of Chromatography A 2015 1388() pp: 251-258
Publication Date(Web):
DOI:10.1016/j.chroma.2015.02.031
Co-reporter:Hui Zhang, Li-Ping Luo, Hui-Peng Song, Hai-Ping Hao, Ping Zhou, Lian-Wen Qi, Ping Li, Jun Chen
Journal of Chromatography A 2014 Volume 1326() pp:47-55
Publication Date(Web):24 January 2014
DOI:10.1016/j.chroma.2013.12.042
•A universal HRPF strategy was developed for discovery of bioactive compounds.•This strategy realized high-purity peak fraction preparation in one HPLC run.•HRPF combined with a luciferase reporter gene assay to screen Nrf2 activators.•20 diterpene quinones were screened out and unambiguously identified.•16 compounds were reported to possess novel Nrf2 activation effect.Generation of a high-purity fraction library for efficiently screening active compounds from natural products is challenging because of their chemical diversity and complex matrices. In this work, a strategy combining high-resolution peak fractionation (HRPF) with a cell-based assay was proposed for target screening of bioactive constituents from natural products. In this approach, peak fractionation was conducted under chromatographic conditions optimized for high-resolution separation of the natural product extract. The HRPF approach was automatically performed according to the predefinition of certain peaks based on their retention times from a reference chromatographic profile. The corresponding HRPF database was collected with a parallel mass spectrometer to ensure purity and characterize the structures of compounds in the various fractions. Using this approach, a set of 75 peak fractions on the microgram scale was generated from 4 mg of the extract of Salvia miltiorrhiza. After screening by an ARE-luciferase reporter gene assay, 20 diterpene quinones were selected and identified, and 16 of these compounds were reported to possess novel Nrf2 activation activity. Compared with conventional fixed-time interval fractionation, the HRPF approach could significantly improve the efficiency of bioactive compound discovery and facilitate the uncovering of minor active components.
Co-reporter:Zi-Qi Shi, De-Fang Song, Ruo-Qian Li, Hua Yang, Lian-Wen Qi, Gui-Zhong Xin, Ding-Qian Wang, Hui-Peng Song, Jun Chen, Haiping Hao, Ping Li
Journal of Chromatography A 2014 Volume 1345() pp:78-85
Publication Date(Web):6 June 2014
DOI:10.1016/j.chroma.2014.04.015
•Qualitative and quantitative analysis of Lycoridis Radiatae Bulbus were performed.•Effective combinatorial markers of Lycoridis Radiatae Bulbus were characterized.•Galanthamine and ungerimine were proved synergy in AChE inhibitory in vitro.•An approach to identifying suitable markers for quality control of HMs was proposed.Quality standardization of herbal medicines (HMs) is an important task with great challenges. Selection of abundant compounds as markers is currently a major approach for the quality control of HMs; however, such marker compounds are irrelevant to the bioactivities in many cases. Taking Lycoridis Radiatae Bulbus (LRB) as an example, we proposed a universal strategy to identify the effective combinatorial markers (ECMs) that are representative of the bioactivities of HMs, and took them as chemical markers for quality standardization. Fingerprinting and quantification were employed to find out the common components in various batches of medicines. The contribution of each common compound to the overall bioactivity was determined through fingerprint–bioactivity modeling, which based on the absolute quantification of each compound and the acetylcholinesterase (AChE) inhibitory activity of LRB. Two most effective compounds, ungerimine and galanthamine, were therefore proposed as ECMs. Interestingly, these two compounds could synergistically inhibit AChE. This approach demonstrated its strong advantage of the bioactivity relevant quality assessment when compared with conventional methods. And the success of applying this ECMs-based method to the quality assessment of unknown LRB samples proved that our approach was reliable and reproducible. In conclusion, this approach is not only useful for the bioactivity relevant quality control of HMs but also helpful for the discovery of ECMs as new drug candidates.
Co-reporter:Li Duan, Long Guo, Ke Liu, E.-Hu Liu, Ping Li
Journal of Chromatography A 2014 Volume 1339() pp:118-127
Publication Date(Web):25 April 2014
DOI:10.1016/j.chroma.2014.02.091
•A novel strategy to characterize and classify seven Citrus herbs based on chromatographic analysis and chemometric methods is developed.•GA-SVM is introduced into classify and predict herbal medicines.•The obtained classifier shows good prediction performance and the overall prediction accuracy reaches 96.88%.•A total of 75 compounds are identified or tentatively characterized in Citrus herbs.•Ten marker compounds are simultaneous quantified within 18 min in seven Citrus herbs.Citrus herbs have been widely used in traditional medicine and cuisine in China and other countries since the ancient time. However, the authentication and quality control of Citrus herbs has always been a challenging task due to their similar morphological characteristics and the diversity of the multi-components existed in the complicated matrix. In the present investigation, we developed a novel strategy to characterize and classify seven Citrus herbs based on chromatographic analysis and chemometric methods. Firstly, the chemical constituents in seven Citrus herbs were globally characterized by liquid chromatography combined with quadrupole time-of-flight mass spectrometry (LC–QTOF-MS). Based on their retention time, UV spectra and MS fragmentation behavior, a total of 75 compounds were identified or tentatively characterized in these herbal medicines. Secondly, a segmental monitoring method based on LC-variable wavelength detection was developed for simultaneous quantification of ten marker compounds in these Citrus herbs. Thirdly, based on the contents of the ten analytes, genetic algorithm optimized support vector machines (GA-SVM) was employed to differentiate and classify the 64 samples covering these seven herbs. The obtained classifier showed good prediction performance and the overall prediction accuracy reached 96.88%. The proposed strategy is expected to provide new insight for authentication and quality control of traditional herbs.
Co-reporter:Long Guo, Li Duan, Ke Liu, E-Hu Liu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2014 Volume 95() pp:220-228
Publication Date(Web):July 2014
DOI:10.1016/j.jpba.2014.03.009
•A RRLC-ESI-MSn method was developed for quantification of 19 compounds in Tripterygium herbs.•Chemometrics methods were performed to compare and discriminate the samples.•The results would be helpful in comparison and evaluation of the quality of Tripterygium herbs.Tripterygium wilfordii (T. wilfordii) and Tripterygium hypoglaucum (T. hypoglaucum), two commonly used Chinese herbal medicines derived from Tripterygium genus, have been widely used for the treatment of rheumatoid arthritis and other related inflammatory diseases in clinical therapy. In the present study, a rapid resolution liquid chromatography/electrospray ionization tandem mass spectrometry (RRLC-ESI-MSn) method has been developed and validated for simultaneous determination of 19 bioactive compounds including four catechins, three sesquiterpene alkaloids, four diterpenoids, and eight triterpenoids in these two similar herbs. The method validation results indicated that the developed method had desirable specificity, linearity, precision and accuracy. Quantitative analysis results showed that there were significant differences in the content of different types of compounds in T. wilfordii and T. hypoglaucum. Moreover, chemometrics methods such as one-way ANOVA, principal component analysis (PCA) and hierarchical clustering analysis (HCA) were performed to compare and discriminate the two Tripterygium herbs based on the quantitative data of analytes, and it was proven straightforward and reliable to differentiate T. wilfordii and T. hypoglaucum samples from different origins. In conclusion, simultaneous quantification of multiple-active component by RRLC-ESI-MSn coupled with chemometrics analysis could be a well-acceptable strategy to compare and evaluate the quality of T. wilfordii and T. hypoglaucum.
Co-reporter:Hua Yang, Lei Liu, Wen Gao, Ke Liu, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2014 Volume 92() pp:13-21
Publication Date(Web):15 April 2014
DOI:10.1016/j.jpba.2013.12.041
•HPLC–QTOF MS method was developed for comprehensive analysis of Shenfu injection.•A total of 44 compounds were characterized by HPLC–QTOF MS.•Twenty-four major alkaloids and ginsenosides were quantified by HPLC–MS.•Similarity analyses for Shenfu injection demonstrated high quality consistency.Shenfu injection (SFI) is a widely used Chinese herbal formulation for cardiac diseases prepared from red ginseng and processed aconite root. Clinical observations and pharmacological effects on SFI have been well investigated. Chemical analysis and quality control studies of this formulation, however, are relatively limited, especially regarding toxic aconite alkaloids. In this work, a high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (HPLC–QTOF MS) method was applied to comprehensive analysis of constituents in SFI. Highly sensitive MS allows direct analysis of injections without additional sample pretreatment required. Using diagnostic ions and fragmentation rules, we identified 23 trace diterpene alkaloids, nineteen ginseng saponins, one panaxytriol, and one 5-hydroxymethylfurfural in SFI. A LC–MS method with selected ion monitoring was then used to quantify 24 major alkaloids and ginsenosides. The method was validated in terms of linearity, accuracy and precision. Especially, the limits of quantification were low to 0.4–18 ng/mL for diterpene alkaloids. The total concentrations of saponins and alkaloids were about 676–742 μg/mL and 3–7 μg/mL in five batches of SFI samples, respectively. Finally, cosine ratio and euclidean distance were introduced to evaluate the batch-to-batch reproducibility of SFI samples, and the results demonstrated high quality consistency. Global identification and quantification of complex constituents based on LC–MS promises wide applications in quality control and batch monitoring for herbal products.
Co-reporter:Xin-Guang Liu, Hua Yang, Xiao-Lan Cheng, Lei Liu, Yong Qin, Qi Wang, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2014 Volume 97() pp:123-128
Publication Date(Web):25 August 2014
DOI:10.1016/j.jpba.2014.04.027
•A MSPD method for extraction and clean-up of ginkgo components.•UHPLC–QQQ-MS for simultaneous analysis of 18 ginkgo components.•Direct quantification of 10 flavonol glycosides without hydrolysis.•Analysis of the 12 Ginkgo biloba tablets available in the global market.Analysis and quality control of Ginkgo biloba have been comprehensively studied. However, little attention has been devoted to the simultaneous extraction and analysis of flavonols and terpene trilactones, especially for direct quantification of flavonol glycosides. This work described a rapid strategy for one-step extraction and quantification of the components. A matrix solid phase dispersion (MSPD) method was designed for the extraction of ginkgo ingredients and compared with the heat-reflux and ultrasonic extraction methods. An ultra-high performance liquid chromatography (UHPLC)–tandem-triple-quadrupole-mass spectrometry (QQQ-MS) method was developed for detection of the 18 components, including 10 original flavonol glycosides, 3 aglycones, and 5 lactones. Subsequently, the proposed strategy was used for the analysis of 12 G. biloba tablets. Results showed that MSPD produced comparable extraction efficiency but consumed less time and required lower solvent volumes compared with conventional methods. Without hydrolysis, the concentration detected was much closer to the original in the sample. The total flavonol glycoside contents in ginkgo tablets ranged from 3.59 to 125.21 μg mg−1, and the terpene trilactone varied from 3.45 to 57.8 μg mg−1 among different manufacturers. In conclusion, the proposed MSPD and UHPLC–QQQ-MS is rapid and sensitive in providing comprehensive profile of chemical constituents especially the genuine flavonol glycosides for improved quality control of ginkgo products.
Co-reporter:Li Duan, Long Guo, Li−Li Dou, Ke-Yun Yu, E-Hu Liu, and Ping Li
Journal of Agricultural and Food Chemistry 2014 Volume 62(Issue 46) pp:11122-11129
Publication Date(Web):October 22, 2014
DOI:10.1021/jf5036355
Citrus grandis ‘Tomentosa’ (CGT) is particularly cultivated in China and widely used in health foods. In this study, the chemical profiles of different parts of CGT were comprehensively compared by rapid resolution liquid chromatography coupled to electrospray ionization quadrupole time-of-flight mass spectrometry method. A total of 22 compounds were identified and two C-glucosyl flavones were found for the first time in CGT. Four main constituents (rhiofolin, naringin, meranzin hydrate, and isoimperatorin) in different parts of CGT were simultaneously determined. Overall, the contents of the four main compounds decreased with the ripening process. In parallel, the antioxidant activities of their extracts were also evaluated by three assays (2,2′-azinobis(3-ethylbenzthiazolinesulfonic acid) diammonium salt, 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power), and the results indicated a similar tendency: small fruit > flower ∼ medium fruit > large fruit > leaf ∼ branch. The results obtained in the present work may provide useful information for future utilization of CGT.
Co-reporter:Wen-Yan Hu, Xue-Jun Kang, Chi Zhang, Jun Yang, Rui Ling, E.-Hu Liu, Ping Li
Journal of Chromatography B 2014 Volume 957() pp:7-13
Publication Date(Web):15 April 2014
DOI:10.1016/j.jchromb.2014.02.036
•A method based on packed-fiber solid-phase extraction (PF SPE) has been established for the determination of three estrogenic stilbenes in dairy products.•Samples were pretreated with PF SPE using an electrospun nanofiber cartridge before analysis by RRLC–MS/MS.•Parameters affecting the efficiency of PF SPE, including pH and salt concentration, were investigated and optimized.•The developed method was successfully applied for the determination of stilbenes in 69 milk samples.•The method has proved to be sensitive and cost-effective in stilbene detection.A sensitive analytical method based on packed-fiber solid-phase extraction and high performance liquid chromatography–tandem mass spectrometry (PF SPE-HPLC–MS/MS) has been developed for determination of three synthetic stilbenes in milk. The stilbenes are extracted with acetonitrile, using sodium chloride, and purified with PF SPE using a cartridge containing electrospun polystyrene nanofibers. Parameters affecting the efficiency of PF SPE, such as pH and amount of salt, were optimized. Under optimal conditions, the limits of detection and quantification were 5–13 pg/g and 15–37 pg/g, respectively. Absolute recoveries varied between 60% and 85% at three different levels. The method was successfully applied for the determination of estrogenic stilbenes in a total of 69 milk samples. The method is sensitive and cost-effective in stilbene detection, and has potential in quality control of dairy products.
Co-reporter:Peng Liu;Hua Yang;Fang Long;Hai-Ping Hao;Xiaojun Xu
Pharmaceutical Research 2014 Volume 31( Issue 7) pp:1788-1800
Publication Date(Web):2014 July
DOI:10.1007/s11095-013-1283-1
To identify bioactive equivalent combinatorial components (BECCs) in herbal medicines. The exact composition of effective components in herbal medicines is often elusive due to the lack of adequate screening methodology. Herein, we propose a hypothesis that BECCs accounting for the whole efficacy of original herbal medicines could be discovered from a complex mixture of constituents.We developed a bioactive equivalence oriented feedback screening method and applied it to discover the BECCs from an herbal preparation Cardiotonic Pill (CP). The operations include chemical profiling of CP, followed by an iterative loop of determining, collecting and evaluating candidate BECCs.A combination of 18 compounds was identified as BECCs from CP, which accounts for 15.0% (w/w) of original CP. We have demonstrated that the BECCs were as effective as CP in cell models and in a rat model of myocardial infarction.This work answers the key question of which are real bioactive components for CP that have been used in clinic for many years, and provides a promising approach for discovering BECCs from herbal medicines. More importantly, the BECCs could be extended to improve quality control of herbal products and inspire an herbal medicines based discovery of combinatorial therapeutics.
Co-reporter:Hui-Peng Song, Hui Zhang, Yu Fu, Hua-yan Mo, Mu Zhang, Jun Chen, Ping Li
Journal of Chromatography B 2014 Volume 961() pp:56-61
Publication Date(Web):15 June 2014
DOI:10.1016/j.jchromb.2014.05.001
•Ultrafiltration LC–MS with enzyme channel blocking was used to screen XOD inhibitors.•Febuxostat was used as an enzyme channel blocker to reduce false positives.•Some flavonoids in Flos Chrysanthemum were screened out as XOD inhibitors.In this study, a new method based on ultrafiltration liquid chromatography-mass spectrometry (UF-LC–MS) combined with enzyme channel blocking (ECB) was developed to discover bioactive components from herbal medicines. Xanthine oxidase (XOD), a critical enzyme for treating gout, was employed as the target protein for screening. By comparing chromatographic profiles of the compounds binding to XOD before and after the ECB experiment, the selective ligands could be distinguished from the non-selective binders. In this experiment, febuxostat bound to the channel entering into the active site of the enzyme and prevented potential ligands from binding. Finally, four compounds, namely, luteolin-7-O-glucoside, apigenin-7-O-glucoside, luteolin and apigenin were screened and identified as the candidate XOD inhibitors based on the ultrafiltration chromatogram of Flos Chrysanthemum, a famous traditional Chinese medicine used in many prescriptions for gout treatment. To verify the compounds screened further, a microplate method was applied to evaluate their enzyme inhibitory activities. The IC50 values of the above 4 compounds were 23.61, 38.80, 1.54 and 1.96 μM, respectively. The structure-function relationship was also estimated according to the in vitro assay. The results were in favor of the hypothesis that the Flos Chrysanthemum extract might be used for gout treatment by inhibiting XOD.
Co-reporter:Long Guo, Li Duan, Xin Dong, Li-Li Dou, Ping Zhou, E-Hu Liu, Ping Li
Journal of Chromatography B 2014 Volume 973() pp:33-38
Publication Date(Web):15 December 2014
DOI:10.1016/j.jchromb.2014.10.008
•A simple and sensitive LC–MS/MS method was developed for the determination of miltirone in rat plasma.•The method was fully validated and successfully applied to pharmacokinetic study of miltirone.•The absolute bioavailability of miltirone was approximately 3.4%.A rapid and sensitive liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed and validated for the quantification of miltirone concentration in rat plasma. The cytotoxic activity of miltirone was firstly evaluated by the MTT assay and compared with other tanshinones. Quantification was carried out on an Agilent triple quadrupole LC–MS system using multiple reaction monitoring (MRM) mode in positive mode. After simple protein precipitation with acetonitrile, the chromatographic separation of miltirone was achieved by using a Waters Symmetry C18 analytical column (2.1 mm × 100 mm, 3.5 μm) with a mobile phase of acetonitrile (A)–water (B) (75:25, v/v) containing 0.5% formic acid. The monitored transitions were set at m/z 283.1 → 223.1 and m/z 361.0 → 232.9 for miltirone and IS, respectively. The calibration curve was linear over the concentration range of 0.5–200 ng/mL with lower limit of quantification of 0.5 ng/mL. The intra- and inter-day accuracy and precision of miltirone were both within acceptable limits. The developed method was successfully applied to a pharmacokinetic study of following oral administration of 20, 40, 60 mg/kg and an intravenous administration of 0.5 mg/kg to rats. The results indicated that miltirone had linear pharmacokinetic properties within the tested dosage range and was poorly absorbed with an absolute bioavailability of approximately 3.4%.
Co-reporter:Jin-Yi Wan, Peng Liu, Huai-You Wang, Lian-Wen Qi, Chong-Zhi Wang, Ping Li, Chun-Su Yuan
Journal of Chromatography A 2013 Volume 1286() pp:83-92
Publication Date(Web):19 April 2013
DOI:10.1016/j.chroma.2013.02.053
American ginseng is a widely used natural product. Ginseng products are usually taken orally, and human intestinal microflora may metabolize ginsenosides. Existing publications report the metabolite fates of ginsenosides. However, investigations on the comprehensive metabolic profile of American ginseng extract are absent because of the chemical complexity and limitation of analytical methods. In this work, we studied the biotransformation and metabolic profile of American ginseng extract by human intestinal microflora. Human fecal microflora was prepared from a healthy Chinese man and then anaerobically incubated with American ginseng sample at 37 °C for 24 h. A rapid and simple liquid–liquid extraction method was used for sample pretreatment. A highly sensitive and selective liquid chromatography/quadrupole time-of-flight mass spectrometry (LC–Q-TOF-MS) method was used to characterize ginsenosides and related metabolites in the reaction samples. The LC–Q-TOF-MS provides superior data quality and advanced analytical capabilities for profiling, identifying, and characterizing complex metabolites in matrix-based biological samples. A total of 25 metabolites were detected, 13 of which were undoubtedly assigned by comparison with reference compounds, and 12 others were tentatively identified. The three most abundant metabolites are 20S-ginsenoside Rg3, ginsenoside F2 and compound K. The main metabolic pathways of ginseng saponins are deglycosylation reactions by intestinal microflora through stepwise cleavage of sugar moieties. Subsequent dehydration reactions also occur. Protopanaxadiol- and oleanane-type triterpenoids are easy to metabolize. The intestinal microbiota may play an important role in mediating the metabolism bioactivity of American ginseng.Highlights► The metabolism of American ginseng extract was studied by human fecal microflora. ► LC–Q-TOF-MS was used for profiling and characterizing complex metabolites. ► Twenty-five ginseng saponin metabolites were identified. ► Ginsenoside Rg3, F2 and compound K are the three most abundant metabolites. ► The main metabolic pathways are deglycosylation and subsequent dehydration.
Co-reporter:Jue Chen, Jun Cao, Wen Gao, Lian-Wen Qi and Ping Li
Analyst 2013 vol. 138(Issue 20) pp:5933-5941
Publication Date(Web):24 Jul 2013
DOI:10.1039/C3AN00957B
Ionic liquids (ILs) have numerous chemical applications as environmentally green solvents that are extending into microemulsion applications. In this work, a novel benign IL-in-water microemulsion system modified by an IL surfactant has been proposed for simultaneous extraction of hydrophilic and lipophilic constituents from Flos Chrysanthemi (Chrysanthemum morifolium). Constituents were analyzed by rapid-resolution liquid chromatography coupled with quadrupole time-of-flight mass spectrometry. A mixture-design approach was used to optimize the IL surfactant and the IL oil phase in the microemulsion system. Microemulsions consisting of 6.0% 1-dodecyl-3-methylimidazolium hydrogen sulfate, 0.1% 1-vinyl-3-methylimidazolium hexafluorophosphate and 93.9% water offered the acceptable extract efficiency that are comparable to or even better than conventional volatile organic solvents. This assay was fully validated with respect to the linearity of response (r2 > 0.999 over two orders of magnitude), precision (intra-RSD < 0.49 and inter-day RSD < 2.21), and accuracy (recoveries ranging from 93.73% to 101.84%). The proposed IL-in-water microemulsion method provided an environmentally friendly alternative for efficient extraction of compounds from Flos Chrysanthemi and could be extended to complex environmental and pharmaceutical samples.
Co-reporter:Xiao-Lan Cheng, Lian-Wen Qi, Qi Wang, Xin-Guang Liu, Besma Boubertakh, Jin-Yi Wan, E-Hu Liu and Ping Li
Analyst 2013 vol. 138(Issue 8) pp:2279-2288
Publication Date(Web):29 Jan 2013
DOI:10.1039/C3AN36732K
In this work, a rapid and simple method based on matrix solid-phase dispersion (MSPD) and ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed. Guge Fengtong preparation (GGFT), a traditional Chinese herbal medicine, was investigated for validation, and eight major constituents were determined including four saponins (protodioscin, protogracillin, pseudoprotodioscin and dioscin) and four gingerols (6-gingerol, 8-gingerol, 10-gingerol and 6-shogaol). Response surface methodology and desirability function were employed to optimize the extraction conditions, such as dispersant, dispersant/sample ratio, solvent concentration, and elution volume, of MSPD. Results showed that MSPD using C18 (1.75 g) as the dispersant material and methanol (89%, v/v) as the eluting solvent (12.00 mL) resulted in a high extraction efficiency. MSPD extraction had the advantages of combining extraction and clean-up in a single step, was less time consuming and required lower solvent volumes compared with conventional methods. Quantification of chemical compounds from GGFT preparations were performed using UPLC-MS/MS in multiple-reaction monitoring mode. The proposed method afforded a low limit of detection ranging from 0.02 to 0.40 ng for saponins and gingerols. For all the analytes, recoveries ranged from 80.9% to 103% and repeatabilities were acceptable with relative standard deviations of less than 6.81%. The proposed MSPD-UPLC-MS/MS method was successfully utilized to analyze five batches of GGFTs, and the results demonstrated that this method is simple, efficient and has potential to be applied for the quality control of herbal preparations.
Co-reporter:G Li, T Zhou, L Liu, J Chen, Z Zhao, Y Peng, P Li and N Gao
Blood Cancer Journal 2013 3(4) pp:e108
Publication Date(Web):2013-04-01
DOI:10.1038/bcj.2013.7
Ezrin links the actin filaments with the cell membrane and has a functional role in the apoptotic process. It appears clear that ezrin is directly associated with Fas, leading to activation of caspase cascade and cell death. However, the exact role of ezrin in ursolic acid (UA)-induced apoptosis remains unclear. In this study, we show for the first time that UA induces apoptosis in both transformed and primary leukemia cells through dephosphorylation/downregulation of ezrin, association and polarized colocalization of Fas and ezrin, as well as formation of death-inducing signaling complex. These events are dependent on Rho-ROCK1 signaling pathway. Knockdown of ezrin enhanced cell death mediated by UA, whereas overexpression of ezrin attenuated UA-induced apoptosis. Our in vivo study also showed that UA-mediated inhibition of tumor growth of mouse leukemia xenograft model is in association with the dephosphorylation/downregulation of ezrin. Such findings suggest that the cytoskeletal protein ezrin may represent an attractive target for UA-mediated lethality in human leukemia cells.
Co-reporter:Yu Fu, Jun Chen, Yan-Jing Li, Yun-Feng Zheng, Ping Li
Food Chemistry 2013 Volume 141(Issue 2) pp:1063-1071
Publication Date(Web):15 November 2013
DOI:10.1016/j.foodchem.2013.03.089
•Six flavonoids were isolated from two Glycyrrhiza species.•Some of the isolates displayed potent antioxidant and/or anti-inflammatory activity.•The antioxidant and/or anti-inflammatory activities of three flavonoids were first reported.•These isolated flavonoids were quantified in four Glycyrrhiza species.Licorice, the roots and rhizomes of several Glycyrrhiza species (Leguminosae), is an important natural sweetening agent and a widely used herbal medicine. In this work, six flavonoids, 5-(1,1-dimethylallyl)-3,4,4′-trihydroxy-2-methoxychalcone (1), licochalcone B (2), licochalcone A (3), echinatin (4), glycycoumarin (5) and glyurallin B (6), were isolated from the extracts of licorice (Glycyrrhiza inflata and Glycyrrhiza uralensis). Their structures were elucidated using various spectroscopic methods. To our knowledge, compound 1 was isolated from natural plants for the first time. All the isolates were tested by antioxidant and anti-inflammatory assays. Compounds 2, 4 and 5 showed strong scavenging activity toward the ABTS+ radical, and compounds 1, 2, 3, 5 and 6 exhibited potent inhibition of lipid peroxidation in rat liver microsomes compared with the reference controls. Compounds 1–4 dose-dependently inhibited LPS induced reactive oxygen species (ROS) production in RAW 264.7 cells. Furthermore, compounds 1–5 were demonstrated to inhibit the production of nitric oxide (NO), interleukin-6 (IL-6) and prostaglandin E2 (PGE2) in LPS-induced macrophage cells. Moreover, the contents of the six compounds, in different Glycyrrhiza species, were quantified by HPLC–MS.
Co-reporter:Wen Gao, Hua Yang, Lian-Wen Qi, E-Hu Liu, Mei-Ting Ren, Yu-Ting Yan, Jun Chen, Ping Li
Journal of Chromatography A 2012 Volume 1245() pp:109-116
Publication Date(Web):6 July 2012
DOI:10.1016/j.chroma.2012.05.027
Plant-based medicines become increasingly popular over the world. Authentication of herbal raw materials is important to ensure their safety and efficacy. Some herbs belonging to closely related species but differing in medicinal properties are difficult to be identified because of similar morphological and microscopic characteristics. Chromatographic fingerprinting is an alternative method to distinguish them. Existing approaches do not allow a comprehensive analysis for herbal authentication. We have now developed a strategy consisting of (1) full metabolic profiling of herbal medicines by rapid resolution liquid chromatography (RRLC) combined with quadrupole time-of-flight mass spectrometry (QTOF MS), (2) global analysis of non-targeted compounds by molecular feature extraction algorithm, (3) multivariate statistical analysis for classification and prediction, and (4) marker compounds characterization. This approach has provided a fast and unbiased comparative multivariate analysis of the metabolite composition of 33-batch samples covering seven Lonicera species. Individual metabolic profiles are performed at the level of molecular fragments without prior structural assignment. In the entire set, the obtained classifier for seven Lonicera species flower buds showed good prediction performance and a total of 82 statistically different components were rapidly obtained by the strategy. The elemental compositions of discriminative metabolites were characterized by the accurate mass measurement of the pseudomolecular ions and their chemical types were assigned by the MS/MS spectra. The high-resolution, comprehensive and unbiased strategy for metabolite data analysis presented here is powerful and opens the new direction of authentication in herbal analysis.Highlights► A comprehensive metabolomic strategy was developed for herbal authentication. ► Non-targeted compounds were analyzed in minutes by molecular feature extraction. ► Multivariate statistical analysis showed good prediction performance. ► Totally 82 markers were obtained from 33-batch samples covering 7 Lonicera species.
Co-reporter:Ying Liu, Xiao-Wei Shi, E-Hu Liu, Long-Sheng Sheng, Lian-Wen Qi, Ping Li
Journal of Chromatography A 2012 Volume 1254() pp:43-50
Publication Date(Web):7 September 2012
DOI:10.1016/j.chroma.2012.07.020
Various analytical technologies have been developed for quantitative determination of marker compounds in herbal medicines (HMs). One important issue is matrix effects that must be addressed in method validation for different detections. Unlike biological fluids, blank matrix samples for calibration are usually unavailable for HMs. In this work, practical approaches for minimizing matrix effects in HMs analysis were proposed. The matrix effects in quantitative analysis of five saponins from Panax notoginseng were assessed using high-performance liquid chromatography (HPLC). Matrix components were found to interfere with the ionization of target analytes when mass spectrometry (MS) detection were employed. To compensate the matrix signal suppression/enhancement, two matrix-matched methods, standard addition method with the target-knockout extract and standard superposition method with a HM extract were developed and tested in this work. The results showed that the standard superposition method is simple and practical for overcoming matrix effects for quantitative analysis of HMs. Moreover, the interference components were observed to interfere with light scattering of target analytes when evaporative light scattering detection (ELSD) was utilized for quantitative analysis of HMs but was not indicated when Ultraviolet detection (UV) were employed. Thus, the issue of interference effects should be addressed and minimized for quantitative HPLC–ELSD and HPLC–MS methodologies for quality control of HMs.Highlights► Accurate quantification is important for quality control of herbal medicines. ► Blank matrix sample is usually unavailable for herbal medicines. ► Standard superposition method was developed for quantitative determination for HMs. ► Matrix components interfered with the ionization and light scattering of analytes. ► Standard superposition method is simple and practical for overcoming matrix effects.
Co-reporter:Cui-Li Li, Jiang Ma, Li Zheng, Hui-Jun Li, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2012 Volume 71() pp:71-78
Publication Date(Web):December 2012
DOI:10.1016/j.jpba.2012.07.031
The emodin-involved hepatotoxicity has been gaining increasing attention. The purpose of the present study was to evaluate the cytotoxicity of emodin on cultured human liver cells (L-02) and predict the possible relation between its cytotoxicity and cellular toxicokinetics. Cell viability and cell damage were assessed by Cell Counting Kit-8 (CCK-8) assay and phase-contrast microscopy, respectively. Cytotoxicity tests demonstrated a concentration- and time-dependent toxic effect of emodin on L-02 cells. Furthermore, emodin at concentration of 30 μM led to a significant apoptosis in a time-dependent manner supported by the morphological changes of drug-treated cells. In addition, to elucidate the toxicokinetic characteristics of emodin, a highly sensitive and selective liquid chromatography–mass spectrometry (LC–MS) method was employed and validated for detecting the dynamic alteration of emodin in cells and cell culture media. The proposed method appeared to be suitable for the analysis of emodin with desirable linearity (r2 > 0.99), and satisfying precision being less than 8.7%. The range of recoveries of this method was 90.2–101.9%. The preliminary cellular toxicokinetic study revealed a time-dependent intracellular accumulation of emodin, which was consistent with its in vitro toxic effects. These findings confirmed the cytotoxicity of emodin against L-02 cells and displayed the cytotoxic manner of emodin in terms of its cellular uptake and accumulation in L-02 cells.
Co-reporter:Yu Fu, Wen Gao, Jun-jie Yu, Jun Chen, Hui-jun Li, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2012 Volumes 64–65() pp:64-71
Publication Date(Web):May–June 2012
DOI:10.1016/j.jpba.2012.02.006
Baccharane glycosides represent a group of rare saponins in plant kingdom and have been regarded as chemical marker for quality control of Impatientis Semen. Based on the structural skeleton, the baccharane glycosides were classified into three types: hosenkol A, hosenkol B and hosenkol C type. In this study, a rapid-resolution liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (RRLC/ESI-Q-TOF MS/MS) was performed to investigate the fragmentation behaviours of baccharane glycosides from Impatientis Semen. In full scan mass spectrum, the accurate determination of molecular formula was obtained by the predominant ion [M+COO]− in negative mode. In the MS/MS spectrum, fragmentation reactions of the [M+H]+ acquired in positive mode were recorded to provide abundant structural information on the aglycone and glycosyl moieties. The characteristic ion for hosenkol A and hosenkol B type glycosides was at m/z 399, while for hosenkol C type glycosides the diagnostic ion was at m/z 381. Neutral losses of monosaccharide, disaccharide, H2O and C3H4 were observed for stepwise structural characterization. As a result, 19 compounds including 9 target saponins and 10 non-target saponins were rapidly screened out in ethanol extract of Impatientis Semen, and 5 of them were found to be novel baccharane glycosides.
Co-reporter:Jiang Ma, Lian-Wen Qi, Hui-Jun Li, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2012 Volume 62() pp:155-161
Publication Date(Web):25 March 2012
DOI:10.1016/j.jpba.2011.11.001
Radix Polygoni Multiflori, Rhizoma et Radix Polygoni Cuspidati, and Radix et Rhizoma Rhei are the most frequently used traditional Chinese medicines in the family Polygonaceae. The three herbal medicines (HMs) contain similar types of compounds. In Chinese Pharmacopoeia, five high-performance liquid chromatography (HPLC) coupled with ultraviolet (UV) detection methods have been employed for their quality control. The aim of this study was to develop a simple and conventional strategy, segmental monitoring based on variable wavelength detection (VWD), for simultaneous quantification of phenolic acids, flavonoids, stilbenes and anthraquinones in the three chemically analogous HMs. Compared with the commonly used HPLC-diode array detection (DAD), the proposed method afforded desirable performance on linearity, precision and sensitivity. Additionally, a quadrupole time-of-flight mass spectrometry (QTOF-MS) was applied to the structural confirmation of analytes from complex matrices. Based on the chemical profiles and contents of the analyses, 27 samples from three HMs were well classified using the principal component analysis. The results of this study demonstrated the potential and applicability of segmental monitoring strategy based on VWD for comprehensive quality control of HMs.
Co-reporter:Rong-Hua Ma, Jie Yang, Lian-Wen Qi, Gui-Zhong Xin, Chong-Zhi Wang, Chun-Su Yuan, Xiao-Dong Wen, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2012 Volume 61() pp:22-29
Publication Date(Web):5 March 2012
DOI:10.1016/j.jpba.2011.11.014
Spinosin, a major bioactive herbal ingredient isolated from Semen Ziziphi Spinosae, plays an important role in sedation and hypnosis. However, the pharmacokinetic behavior of spinosin in special sites has not been reported. Microdialysis (MD) technique, as a continuous, realtime monitoring sampling technique, is very suitable for the evaluation of the disposition of diverse drugs. To obtain more useful information on spinosin, an in vivo microdialysis sampling technique with High Performance Liquid Chromatography–mass spectrograph (HPLC–MS) method was developed to investigate the pharmacokinetics of spinosin and its interaction with cyclosporin A (CsA) in the brain, blood and bile of rats. The method was validated in terms of selectivity, linearity and sensitivity, and showed advantages in monitoring the pharmacokinetic behavior of drugs. The results revealed that CsA has obvious effects on the pharmacokinetic process of spinosin. When co-administered, the area under the curve (AUC) of spinosin in blood, bile and brain increased from 205.70 to 673.51 mg min/L, 7.77 × 104 to 1.25 × 105 mg min/L, and 2.09 to 5.58 mg min/L, respectively. The t1/2 values of spinosin in blood, bile and brain also changed from 48.07 to 95.04 min, from 97.20 to 152.21 and from 42.18 to 73.83 min, respectively. These results demonstrated that the CsA decreased the efflux of spinosin through the inhibition of P-glycoprotein (P-gp) efflux transporter and it might be used as a group of P-gp substrate. Other transporters or pathways may also be involved in the metabolism of spinosin.Highlights► Analysis of hepatobiliary excretion and brain metabolism is important to understanding drug metabolism. ► Technical difficulties limited spinosin analysis in rat blood, bile and brain. ► A microdialysis technique with HPLC–MS method was developed to investigate the pharmacokinetics of unbound spinosin in the brain, blood and bile dialysate of rats, this method can be conveniently and sensitively applied to monitor the pharmacokinetic behavior of drugs.
Co-reporter:Xiao-Dong Wen, E-Hu Liu, Jie Yang, Chang-Yin Li, Wen Gao, Lian-Wen Qi, Chong-Zhi Wang, Chun-Su Yuan, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2012 Volumes 67–68() pp:114-122
Publication Date(Web):August–September 2012
DOI:10.1016/j.jpba.2012.04.026
In the present study, rapid resolution liquid chromatography was coupled with quadrupole time-of-flight tandem mass spectrometry (RRLC–Q-TOF–MS) to identify the absorbed components and metabolites in rat urine after oral administration of Buyang Huanwu decoction (BYHWD). After oral administration of BYHWD, urine samples were collected and pretreated by solid phase extraction. The mass measurements were accurate within 5 ppm of error for all the protonated molecules, and subsequent fragment ions offered higher quality structural information for interpretation of the fragmentation pathways of various compounds. A total of 50 compounds were detected in rat urine samples within 20 min, including 12 parent compounds and 38 metabolites. Except for three prototype components (Hydroxysafflor yellow A, Paeoniflorin, and Amygdalin), the metabolites identified mainly came from Radix Astragali, Radix Angelicae Sinensis, and Rhizoma Chuanxiong. The results indicated that glucuronidation and sulfation were the major metabolic pathways of isoflavonoids, while glutathione conjugation, glucuronidation and sulfation were the main metabolic pathways of phthalides. No saponin-related metabolites were detected. The present study provided important structural information on the metabolism of BYHWD. Furthermore, the results of this work have demonstrated the feasibility of the RRLC/ESI-Q-TOF–MS approach for rapid and reliable characterization of metabolites from herbal medicines.
Co-reporter:Gui-Zhong Xin, Liu Cao, Zi-Qi Shi, Hui-Jun Li, Xiao-Dong Wen, Jun Chen, Lian-Wen Qi, Ping Li
Journal of Chromatography B 2012 Volume 899() pp:127-134
Publication Date(Web):15 June 2012
DOI:10.1016/j.jchromb.2012.05.012
Sample pretreatment is a key step in bioanalytical process because of possible interference and matrix effects in mass spectrometry analysis. In this work, a novel strategy towards high speed and sensitivity was developed combining in vivo microdialysis (MD) sampling, turbulent-flow chromatography (TFC), and liquid chromatography–mass spectrometry (LC–MS). The procedures of cleanup, preconcentration, and separation were completed on-line in one step within 10 min. During the MD optimization procedure, 1% hydroxypropyl-β-cyclodextrin (HP-β-CD) was used to improve the relative recovery of the analyte. Untreated MD samples were directly injected, and a TFC precolumn was flushed for 1 min with aqueous phase of 4 mL/min flow rate to desalt and concentrate biosamples. The retained analytes were then back-flushed by a switching valve onto a fast LC column (4.6 mm × 50 mm, 1.8 μm) for separation. Another diverter valve was employed to prevent the HP-β-CD that interferes with the ESI process from entering the MS. Puqietinone, a lipophilic alkaloid from Fritillaria puqiensis, was used as a case for validation. Results showed that the limit of quantification for puqietinone was 0.10 ng/mL, and good linearity (R2 = 0.9993) was maintained over the range of 1.02–200.02 ng/mL. Accuracy and precision were satisfactory within the range of the standard curve. This approach was able to effectively eliminate the influences of matrix effect and carry-over as the injection volume increased up to 20 μL. The developed method was successfully applied to pharmacokinetic study of puqietinone after intravenous administration to rat. Results demonstrate the potential of using MD with TFC-LC/MS for in vivo monitoring experiments.Highlights► Microdialysis sampling and turbulent-flow chromatography were coupled to LC/MS. ► The system is able to achieve online desalting and concentrating within 1 min. ► Influences of matrix effect and carry-over were eliminated effectively. ► Lifetime of extraction column is much extended. ► The temporal resolution of sampling points is improved by using MD technology.
Co-reporter:Hui Zhang, Shao-Qing Wang, Ying Liu, Li-Ping Luo, Peng Liu, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2012 70() pp: 169-177
Publication Date(Web):
DOI:10.1016/j.jpba.2012.06.025
Co-reporter:Xiao-Lan Cheng, Qun Liu, Yong-Bo Peng, Lian-Wen Qi, Ping Li
Food Chemistry 2011 Volume 129(Issue 4) pp:1785-1792
Publication Date(Web):15 December 2011
DOI:10.1016/j.foodchem.2011.06.026
Ginger, from the rhizome of Zingiber officinale Rosco (Zingiberaceae), is a common condiment for foods and beverages. In this work, we tested a hypothesis that a steaming process affects the chemical profile and anticancer potential of ginger. An HPLC method with TOF/MS and DAD was developed to analyse the chemical constituents in ginger. The antiproliferative effect of fresh, dried and steamed gingers was evaluated using human Hela cancer cells. The results showed that the antiproliferative effect of steamed ginger at 120 °C for 4 h was approximately 1.5- and 2-fold higher than that of dried and fresh ginger, respectively. Twenty-two components were characterised in the steamed ginger. The decreased concentration of gingerols and increased levels of shogaols contributed to the improved anticancer potential of the steamed ginger. This study elucidated the relationship of the heating process with the constituents and anticancer activity, and developed an optimised processed ginger extract for chemoprevention.Highlights► The constituents in treated and untreated ginger were compared by HPLC-DAD–TOF/MS. ► The steaming process could enhance the anticancer effects of ginger. ► The increased level of shogoals contributed to the improved anticancer potential.
Co-reporter:Xiao-Lan Cheng, Jin-Yi Wan, Ping Li, Lian-Wen Qi
Journal of Chromatography A 2011 Volume 1218(Issue 34) pp:5774-5786
Publication Date(Web):26 August 2011
DOI:10.1016/j.chroma.2011.06.091
Spatholobus suberectus is a widely used herb in traditional medicine for the treatment of blood stasis syndrome and related diseases. In this work, a potential ultrasonic/microwave assisted extraction (UMAE) method was developed for efficient sample pretreatment, and a diagnostic ion filtering strategy with liquid chromatography–quadrupole time-of-flight mass spectrometry (LC–Q-TOF-MS) was established for rapid characterization of flavonoids in S. suberectus. The factors of UMAE influencing the extraction yield of flavonoids of S. suberectus were evaluated. The optimal conditions were determined as: microwave power of 300 W, extraction time of 450 s, 70% methanol as extraction solvent, solvent to solid ratio of 20 mL/g, ultrasound power of 50 W, extraction temperature of 80 °C, and one extraction cycle. Compared with commonly used extraction methods, UMAE showed higher efficiency and shorter extraction time for sample preparation. Subsequently, the major diagnostic ions and fragmentation pathways of flavonoids in Q-TOF-MS were summarized with available reference compounds. Using a new diagnostic ion filtering strategy, a rapid screening and identification of thirty-eight compounds was achieved in real S. suberectus samples. The results of this study clearly demonstrate the potential of UMAE for efficient extraction and LC–Q-TOF-MS for rapid and sensitive structural elucidation of flavonoids in S. suberectus, and open perspectives for similar studies on other medicinal herbs.
Co-reporter:Yan-Jing Li, Jun Chen, Ying Li, Qin Li, Yun-Feng Zheng, Yu Fu, Ping Li
Journal of Chromatography A 2011 Volume 1218(Issue 45) pp:8181-8191
Publication Date(Web):11 November 2011
DOI:10.1016/j.chroma.2011.09.030
Licorice, derived from the dried roots and rhizomes of several species of genus Glycyrrhiza L. (Leguminosae family), has been traditionally used in herbal medicine for over 4000 years. In recent years, the interest in antioxidative constituents in licorice has greatly increased. In this work, a new method based on 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) spiking test combined with HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF MS/MS) analysis was developed to screen and identify the antioxidants in licorice. The results of the method validation indicated that the developed method was reliable and repeatable. Compared with DPPH on-line method, the HPLC–Q-TOF MS/MS method combined with DPPH spiking test offered much higher sensitivity and resolution. Using this method, 35 radical scavengers were screened from four Glycyrrhiza species (G. inflata, G. glabra, G. pallidiflora and G. uralensis), and 21 of them were unambiguously or tentatively identified by HPLC–Q-TOF MS/MS. Among the 21 identified flavonoids, 10 compounds had been reported to possess antioxidative activities in the previous studies, and the radical scavenging activities of the other 11 compounds were reported for the first time. The effects of six purified flavonoids on DPPH radical and lipid peroxidation were evaluated for validation of the developed method. The results indicated that HPLC–Q-TOF MS/MS coupled with DPPH treatment is an efficient and powerful method to discover the potential antioxidative compounds from the complex natural product mixtures. In this study, the identified components with free radical scavenging activity, would help to explain the therapeutic benefit of licorice in the treatment of human disease associated with oxidative stress.Highlights► DPPH spiking test combined with HPLC–Q-TOF MS/MS to screen antioxidants. ► Higher sensitivity and resolution compared with DPPH on-line method. ► 21 radical scavengers in licorice were identified and 11 of them were first reported.
Co-reporter:Jun Cao, Ping Li, Ling Yi
Journal of Chromatography A 2011 Volume 1218(Issue 52) pp:9428-9434
Publication Date(Web):30 December 2011
DOI:10.1016/j.chroma.2011.11.013
A new CE system using ionic liquids coated multi-walled carbon nanotubes (ILs-MWNTs) as pseudostationary phase was developed for the simultaneous determination of four flavonoids, four phenolic acids and two saponins. Several parameters affecting the separation were studied, including the choice of ILs, ILs-MWNTs concentration, the respective use of ILs and MWNTs, buffer pH, SDS concentration and borate content. Results revealed that the addition of ILs-MWNTs in running electrolytes enhanced the separation of target compounds compared to conventional micelle because the surface of carbon nanotubes interacted favorably with the analytes. Under the optimum conditions, a baseline separation was achieved for these analytes within 11 min in a 41.5 cm of effective length fused-silica capillary. At a voltage of 28.0 kV, the separation was carried out in a 10 mM borate buffer (pH 9.0) containing 100 mM SDS, 6% propanol and 4 μg mL−1 ILs-MWNTs. All calibration curves showed good linearity (r2 > 0.9990) within the test ranges. The intra- and inter-day precisions as determined from standard solutions were below 3.30% and 6.23%, respectively. The recoveries for ten compounds were found to range from 85.5 to 101.8%. The method was successfully applied for the determination of three types of compounds in Qishenyiqi dropping pills. Our experimental results indicated that the proposed method offered new opportunities for the analysis of complex samples.Highlights► A novel CE system based on ILs-MWNTs was reported. ► The pseudostationary phase interacted favorably with analytes. ► The effect of ILs, MWNTs, pH, SDS and borate concentrations was investigated. ► The method was then applied to a determination of Qishenyiqi dropping pills.
Co-reporter:E-Hu Liu;Qun Liu;Chu Chu
Journal of Separation Science 2011 Volume 34( Issue 19) pp:2566-2575
Publication Date(Web):
DOI:10.1002/jssc.201100254
Abstract
A fast high-performance liquid chromatography (HPLC) method with diode-array detection (DAD) and time-of-flight mass spectrometry (TOF/MS) has been developed for the analysis of multi-constituent in Yinhuang granules, a well-known combined herbal remedy prepared from the extract mixtures of Flos Lonicerae and Radix Scutellariae. The fast HPLC analysis was performed on an Agilent ZorBax SB-C18 column (4.6×50 mm, 1.8 μm) and 0.2% aqueous formic acid and acetonitrile was the optimum mobile phase for gradient elution in 17 min, which is five times faster than the performance of conventional columns packed with 5.0 μm particles. With various fragmentor voltages in TOF/MS, accurate mass measurements (<5 ppm error) for molecular ions and characteristic fragment ions represented reliable identification criteria for different constituents. A total of 28 compounds, including nine phenolic acids, three iridoid glycosides and nine saponins from Flos Lonicerae and seven flavonoids from Radix Scutellariae, were identified or tentatively characterized in the extract of Yinhuang granules. The established fast HPLC-DAD-TOF/MS method turns out to be useful and efficient for quality control of this commonly used Chinese herbal preparation.
Co-reporter:Hui-Jun Li, Jun-Jie Yu, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2011 54(4) pp: 674-680
Publication Date(Web):
DOI:10.1016/j.jpba.2010.10.014
Co-reporter:Chang-Yin Li, Lian-Wen Qi, Ping Li
Journal of Pharmaceutical and Biomedical Analysis 2011 55(1) pp: 146-160
Publication Date(Web):
DOI:10.1016/j.jpba.2010.12.034
Co-reporter:Jian-Liang Zhou, Gui-Zhong Xin, Zi-Qi Shi, Mei-Ting Ren, Lian-Wen Qi, Hui-Jun Li, Ping Li
Journal of Chromatography A 2010 Volume 1217(Issue 45) pp:7109-7122
Publication Date(Web):5 November 2010
DOI:10.1016/j.chroma.2010.09.019
Liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (LC/ESI-QTOF-MS/MS) was performed to study the fragmentation behaviors of steroidal alkaloids from Fritillaria species, the antitussive and expectorant herbs widely used in traditional Chinese medicine. We propose, herein, a strategy that combining key diagnostic fragment ions and the relative abundances and amounts of major fragment ions (the ions exceeding 10% in abundance) to distinguish different sub-classes of Fritillaria alkaloids (FAs). It was found that hydrogen rearrangement and induction effects result in ring cleavage of the basic skeletons occurred in the MS/MS process and produced characteristic fragment ions, which are useful for structural elucidation. This method was finally used to investigate the primary steroidal alkaloids in the extracts of eight major Fritillaria species. As a result, 41 steroidal alkaloids (29 cevanine type, 1 jervine type, 6 veratramine type and 5 secosolanidine type alkaloids) were selectively identified in these Fritillaria species. Twenty-six compounds were unambiguously identified by comparing with the reference compounds and 15 compounds were tentatively identified or deduced according to their MS/MS data. Logical fragmentation pathways for different types of FAs have been proposed and are useful for the identification of these types of steroidal alkaloids in natural products especially when there are no reference compounds available.
Co-reporter:Ying Liu, Jian-Liang Zhou, Peng Liu, Shi Sun, Ping Li
Journal of Chromatography A 2010 Volume 1217(Issue 32) pp:5239-5245
Publication Date(Web):6 August 2010
DOI:10.1016/j.chroma.2010.06.039
A strategy based on chemical markers’ fishing and knockout has been proposed for holistic activity and interaction evaluation of the bioactive components in herbal medicines (HMs). It was devised to screen bioactive-compound group that represents the efficacy of HM, estimate the bioactivity contribution of each component and elucidate the interactions of multi-components. This strategy was accomplished through the following steps: (1) screen out the chemical markers (target peaks) in a HM fingerprint using online two-dimensional turbulent flow chromatography/liquid chromatography–mass spectrometry technique, (2) fish target peaks and knockout any interested peak, and (3) evaluate the bioactivities of fishing and knockout portions. After comparison of the bioactivities of samples containing different target peaks, the efficacy of target-peak group, bioactivity contribution of each compound, and the interactions of multi-components are elucidated. Using Acetylcholinesterase (AChE) and Bulbs of Lycoris radiata (L. Herit.) Herb. (BLR) as the experimental materials, four target peaks were screened out as the AChE binders. By target peaks’ fishing and knockout, combined with activity evaluation, we observed that the bioactivity of the four-peak mixture is similar with the global bioactivity of BLR extract, and there are significant suppressive actions among these four target peaks. These results indicate that this proposed strategy is a useful approach for holistic screening of bioactive-compound group and elucidation of the multi-component interactions in HM.
Co-reporter:Gui-Zhong Xin, Jian-Liang Zhou, Lian-Wen Qi, Chang-Yin Li, Peng Liu, Hui-Jun Li, Xiao-dong Wen, Ping Li
Journal of Chromatography B 2010 Volume 878(3–4) pp:435-441
Publication Date(Web):1 February 2010
DOI:10.1016/j.jchromb.2009.12.027
A method based on the on-line turbulent-flow chromatography and fast high-performance liquid chromatography/mass spectrometry (TFC–LC/MS) was developed for sensitive and high throughput pharmacokinetic study of traditional Chinese medicines (TCMs). In this method, an on-line extraction column (Waters Oasis HLB) and a fast HPLC column with sub-2 μm particle size (Agilent Zorbax StableBond-C18, 4.6 mm × 50 mm, 1.8 μm) in a column-switching set-up were utilized. HLB is a reversed-phase extraction column with hydrophilic–lipophilic balanced copolymer (2.1 mm × 20 mm, 25 μm particle size), which will exhibit some turbulent-flow properties at a high-flow rate. The method combines the speed and robustness of turbulent-flow extraction and the sensitivity and separation efficiency of fast HPLC–MS to analyze multiple and trace constituents of TCMs in plasma matrix. This method was successfully applied for pharmacokinetic study of verticine, verticinone and isoverticine, the chemical markers of Fritillaria thunbergii, after oral administration of total steroidal alkaloids extract of F. thunbergii to rats. Each plasma sample was analyzed within 7 min. The method demonstrated good linearity (R > 0.999) ranged from 0.505 to 96.0 ng/mL with satisfactory accuracy and precision, and the lower limit of quantifications of verticine, verticinone and isoverticine were estimated to be 0.120, 0.595 and 0.505 ng/mL, respectively. These results indicate that the proposed method is fast, sensitive, and feasible for pharmacokinetic study of TCMs.
Co-reporter:Jun Cao, Ling Yi, Ping Li, Yan-xu Chang
Journal of Chromatography A 2009 Volume 1216(Issue 29) pp:5608-5613
Publication Date(Web):17 July 2009
DOI:10.1016/j.chroma.2009.05.060
To separate and detect neutral solutes in nonionic microemulsion electrokinetic chromatography (MEEKC), a novel method was developed, combining complex formation and acetonitrile (ACN) sweeping. In this report, dynamic borate complexation and on-line sweeping occurred simultaneously during a run. The operating parameters which affected the performance of analyte sweeping in nonionic MEEKC were examined in terms of borate complexation, ACN content, Brij-35 concentration and sample plug length. In addition, the validation of the method included tests of the limit of detection, reproducibility and sensitivity enhancement. 60–110-Fold of magnitude improvement in detection sensitivity for model compounds (ginsenoside Rf, ginsenoside Rb2, ginsenoside Re) using Brij-35 microemulsion was demonstrated. Furthermore, the method was applied to the determination of glucosides in the plant extract.
Co-reporter:Hui-Jun Li, Yan Jiang, Ping Li
Journal of Chromatography A 2009 Volume 1216(Issue 11) pp:2142-2149
Publication Date(Web):13 March 2009
DOI:10.1016/j.chroma.2008.03.093
Bulbus Fritillariae (BF), referred to the bulbs of several Fritillaria species (Liliaceae), is a commonly used antitussive and expectorant herb in traditional Chinese medicine (TCM). Due to the complexity of BF botanical origin in the herbal markets, it is urgently needed to develop a reliable method for species identification. Previous studies based on morphological and histological as well as molecular biological techniques have respective limitations. For the purpose of finding a possible discriminant method among Fritillaria species, 27 steroidal alkaloids in 17 Fritillaria species and 12 BF-containing compound formulas were identified and characterized by a high-performance liquid chromatography with mass spectrometry (HPLC–MS) method. The estimated relative composition of steroidal alkaloids was used to carry out a chemotaxonomical study on Fritillaria species by means of hierarchial cluster analysis. In addition, the characteristic occurring patterns of the examined bases were compared in an effort to distinguish the botanical origin of BF-containing compound formulas. The results demonstrated that the qualitative and quantitative differences in steroidal alkaloids were useful not only for chemotaxonomy in some medicinal Fritillaria species but also for species identification in compound formulas. The described method has important implications in quality control of BF-containing TCM preparations, allowing for the prevention of BF confusion, and also revealing the possible presence of adulteration.
Co-reporter:Jian-Liang Zhou, Jing-Jing An, Ping Li, Hui-Jun Li, Yan Jiang, Jie-Fei Cheng
Journal of Chromatography A 2009 Volume 1216(Issue 12) pp:2394-2403
Publication Date(Web):20 March 2009
DOI:10.1016/j.chroma.2009.01.010
We present herein a novel bioseparation/chemical analysis strategy for protein–ligand screening and affinity ranking in compound mixtures, designed to increase screening rates and improve sensitivity and ruggedness in performance. The strategy is carried out by combining on-line two-dimensional turbulent flow chromatography (2D-TFC) with liquid chromatography–mass spectrometry (LC–MS), and accomplished through the following steps: (1) a reversed-phase TFC stage to separate the protein/ligand complex from the unbound free molecules, (2) an on-line dissociation process to release the bound ligands from the complexes, and (3) a second mixed-mode cation-exchange/reversed-phase TFC stage to trap the bound ligands and to remove the proteins and salts, followed by LC–MS analysis for identification and determination of the binding affinities. The technique can implement an ultra-fast isolation of protein/ligand complex with the retention time of a complex peak in about 5 s, and on-line prepare the “clean” sample to be directly compatible with the LC–MS analysis. The improvement in performance of this 2D-TFC/LC–MS approach over the conventional approach has been demonstrated by determining affinity-selected ligands of the target proteins acetylcholinesterase and butyrylcholinesterase from a small library with known binding affinities and a steroidal alkaloid library composed of structurally similar compounds. Our results show that 2D-TFC/LC–MS is a generic and efficient tool for high-throughput screening of ligands with low-to-high binding affinities, and structure-activity relationship evaluation.
Co-reporter:Xiao-Jie Gu;Nan Yao;Shi-Hui Qian;You-Bin Li
Helvetica Chimica Acta 2009 Volume 92( Issue 1) pp:88-97
Publication Date(Web):
DOI:10.1002/hlca.200800186
Co-reporter:Wei Wang, Chang-Yin Li, Xiao-Dong Wen, Ping Li, Lian-Wen Qi
Journal of Pharmaceutical and Biomedical Analysis 2009 49(4) pp: 1070-1074
Publication Date(Web):
DOI:10.1016/j.jpba.2009.01.020
Co-reporter:Wei Wang, Chang-Yin Li, Xiao-Dong Wen, Ping Li, Lian-Wen Qi
Journal of Chromatography B 2009 Volume 877(8–9) pp:671-679
Publication Date(Web):15 March 2009
DOI:10.1016/j.jchromb.2009.01.021
A rapid high-performance liquid chromatography–mass spectrometry (HPLC–MS) method was developed and validated for simultaneous quantification of 6-gingerol, 8-gingerol, 10-gingerol and 6-shogaol in rat plasma after oral administration of ginger oleoresin. Plasma samples extracted with a liquid–liquid extraction procedure were separated on an Agilent Zorbax StableBond-C18 column (4.6 mm × 50 mm, 1.8 μm) and detected by MS with electrospray ionization interface in positive selective ion monitoring (SIM) mode. Calibration curves (1/x2 weighted) offered satisfactory linearity (r2 > 0.995) in a wide linear range (0.0104–13.0 μg/mL for 6-gingerol, 0.00357–4.46 μg/mL for 8-gingerol, 0.00920–11.5 μg/mL for 10-gingerol and 0.00738–9.22 μg/mL for 6-shogaol). The lower limit of quantification (LLOQ) was in a range of 3.57–10.4 ng/mL. The analytes and internal standard can be baseline separated within 6 min. Inter- and intra-day assay variation was less than 15%. This developed method was successfully applied to pharmacokinetic studies of ginger oleoresin after oral administration to rats. Glucuronide of 6-gingerol was determined after β-glucuronidase hydrolysis for more information, and the intestinal glucuronidation was further confirmed by comparison of plasma samples of hepatic portal vein and femoral vein.
Co-reporter:Qian Wu, Xiao-Dong Wen, Lian-Wen Qi, Wei Wang, Ling Yi, Zhi-Ming Bi, Ping Li
Journal of Chromatography B 2009 Volume 877(8–9) pp:751-756
Publication Date(Web):15 March 2009
DOI:10.1016/j.jchromb.2009.02.001
Harpagoside, a major bioactive iridoid glucoside in genus Scrophularia, has been widely used in clinical practice for the treatment of pain in the joints and lower back for its neuroprotective and anti-inflammation activities. To investigate the pharmacokinetics and hepatobiliary excretion, an in vivo microdialysis method coupled with high performance liquid chromatography was developed to monitor the concentration of harpagoside in blood and bile. The harpagoside bile-to-blood distribution ratio (AUCbile/AUCblood) up to 986.28 ± 78.46 significantly decreased to 6.41 ± 0.56 or 221.20 ± 18.92 after co-administration of cyclosporin A or verapamil. The results indicated that harpagoside went through concentrative elimination from the bile which was probably regulated by P-glucoprotein, providing possible clinical trials of co-administration of transporter inhibitors to decrease drug efflux, thus to enhance the curative effects.
Co-reporter:Chun-Yun Chen, Lian-Wen Qi, Ling Yi, Ping Li, Xiao-Dong Wen
Journal of Chromatography B 2009 Volume 877(Issue 3) pp:159-165
Publication Date(Web):15 January 2009
DOI:10.1016/j.jchromb.2008.11.043
A liquid chromatography-electrospray ionization–mass spectrometry method has been developed and validated for identification and quantification of four major bioactive saponins in rat plasma after oral administration of extraction of saponins from Flos Lonicerae, i.e., macranthoidin B, macranthoidin A, dipsacoside B, and macranthoside B. Plasma samples were extracted with solid-phase extraction, separated on a Shim-pack CLC-ODS column and detected by MS in negative selective ion monitoring mode. Calibration curves offered linear ranges of two orders of magnitude with r2 > 0.999. The method showed the low limit quantification of 7.72, 6.06, 7.16, and 1.43 ng/mL for macranthoidin B, macranthoidin A, dipsacoside B, and macranthoside B, respectively. The inter- and intra-CV precision (R.S.D.) were all within 10% and accuracy (% bias) ranged from −10 to 10%. The overall recovery was more than 70%. This developed method was subsequently successfully applied to pharmacokinetic profiles of the four saponins in rats. After oral administration of extraction of saponins in rats, the concentration–time course was found to be the double peaks of curve.
Co-reporter:Lian-Wen Qi, Jun Cao, Ping Li, Yu-Xia Wang
Journal of Pharmaceutical and Biomedical Analysis 2009 49(2) pp: 502-507
Publication Date(Web):
DOI:10.1016/j.jpba.2008.10.026
Co-reporter:Xiao-Dong Wen, Lian-Wen Qi, Bin Li, Ping Li, Ling Yi, Ya-Qiong Wang, E-Hu Liu, Xiao-Lin Yang
Journal of Pharmaceutical and Biomedical Analysis 2009 50(1) pp: 100-105
Publication Date(Web):
DOI:10.1016/j.jpba.2009.03.038
Co-reporter:Jian-Liang Zhou, Ping Li, Hui-Jun Li, Yan Jiang, Mei-Ting Ren, Ying Liu
Journal of Chromatography A 2008 Volume 1177(Issue 1) pp:126-137
Publication Date(Web):4 January 2008
DOI:10.1016/j.chroma.2007.11.030
Steroidal alkaloids are naturally occurring nitrogen-containing compounds in many edible or medicinal plants, such as potato, tomato, Fritillaria and American hellebore, which possess a variety of toxicological and pharmacological effects on humans. The aim of this study is to explore the potential of liquid chromatography/electrospray ionization time-of-flight mass spectrometry (LC/ESI-TOF-MS) method in the determination of these important alkaloids in plant matrices. The application of this method has been proven through 26 naturally occurring steroidal alkaloids in Fritillaria species. Accurate mass measurements within 4 ppm error were obtained for all the alkaloids detected out of various plant matrices, which allowed an unequivocal identification of the target steroidal alkaloids. The bunching factor for mass spectrometer, an important parameter significantly affecting the precision and accuracy of quantitative method, was firstly optimized in this work and satisfactory precision and linearity were achieved by the optimization of that parameter. The ranges of RSD values of intra-day and inter-day variability for all alkaloids were decreased remarkably from 41.8–159% and 13.2–140% to 0.32–7.98% and 2.37–16.1%, respectively, when the value of bunching factor was optimized from 1 to 3. Linearity of response more than two orders of magnitude was also demonstrated (regression coefficient >0.99). The LC/TOF-MS detection method offered improvements to the sensitivity, compared with previously applied LC (or GC) methods, with limits of detection down to 0.0014–0.0335 μg/ml. The results in this paper illustrate the robustness and applicability of LC/TOF-MS for steroidal alkaloids analysis in plant samples. In addition, relative quantitative determination of steroidal alkaloid with one popular analyte verticinone which is commercially available was also investigated in order to break through the choke point of lack of standards in phytochemical analysis. The accuracies of relative quantitative method for steroidal alkaloids determinations with verticinone were 90.6–110.0% (average 98.5%) suggesting that it is feasible to quantify steroidal alkaloids by the proposed relative quantitative determination method within acceptable errors.
Co-reporter:Dan Tang;Hui-Jun Li;Jun Chen;Chao-Wei Guo
Journal of Separation Science 2008 Volume 31( Issue 20) pp:3519-3526
Publication Date(Web):
DOI:10.1002/jssc.200800173
Abstract
A rapid and simple method has been developed for the screening and identification of natural antioxidants of Flos Lonicerae Japonicae (FLJ), derived from the flower buds of Lonicera japonica. The hypothesis is that upon reaction with 1,1-diphenyl-2-picrylhydrazyl (DPPH), the peak areas (PAs) of compounds with potential antioxidant effects in the HPLC chromatograms will be significantly reduced or disappeared, and the identity confirmation could be achieved by HPLC-DAD-TOF/MS hyphenated technique. Using the proposed approach, about 14 compounds in the FLJ extract were found to possess a potential antioxidant activity. They were identified as chlorogenic acid (1), 1-O-caffeoylquinic acid (1-O-CQA, 2), caffeic acid (4), 4-O-CQA (5), rutin (7), isoquercitrin (8), luteolin-7-O-glucoside (9), lonicerin (10), 4,5-O-dicaffeoylquinic acid (4,5-O-diCQA, 11), 3,5-O-diCQA (12), 1,3-O-diCQA (13), 3,4-O-diCQA (14), 1,4-O-diCQA (16), and luteolin (17). In addition, the free radical scavenging capacities of the available identified compounds were also investigated by HPLC assay. The results indicated that the compounds with PAs significantly decreasing were natural antioxidants, whereas those with PAs not changing presented no activities, which accordingly indicated that this newly proposed method could be widely applied for rapid screening and identification of natural antioxidants from complex matrices including Chinese herbal medicines.
Co-reporter:Lian-Wen Qi;Qing-Tao Yu;Ling Yi;Mei-Ting Ren;Xiao-Dong Wen;Yu-Xia Wang
Journal of Separation Science 2008 Volume 31( Issue 1) pp:97-106
Publication Date(Web):
DOI:10.1002/jssc.200700286
Abstract
An improved quality control method was developed to simultaneously determine 15 major constituents (eight flavonoids and seven saponins) in various Radix Astragali preparations, using SPE for pretreatment of samples, HPLC with diode-array and evaporative light scattering detectors (DAD-ELSD) for quantification in one run, and HPLC-ESI-TOF/MS for definite identification of compounds in preparations. Optimum separations were obtained with a ZORBAX C18 column, using a gradient elution with 0.3% aqueous formic acid and ACN. This established method was fully validated with respect to linearity, precision, repeatability, and accuracy, and was successfully applied to quantify the 15 compounds in 19 commercial samples, including 3 dosage forms, i. e., oral solution, injection, concentrated granule, and its processed products of Radix Astragali. The results demonstrated that many factors might result in significant differences in quality of the final preparations, including crude drugs, pretreatment processes, manufacturing procedure, storage conditions, etc. Then the developed method provided a reasonable and powerful manner to ensure the efficacy, safety, and batch-to-batch uniformity of Radix Astragali products by standardizing each procedure, and thus should be proposed as quality control for the clinical use and modernization of herbal preparations.
Co-reporter:Jing Zhao;Qing-Tao Yu, ;Peng Zhou;Yuan-Jie Zhang ;Wei Wang
Journal of Separation Science 2008 Volume 31( Issue 2) pp:255-261
Publication Date(Web):
DOI:10.1002/jssc.200700379
Abstract
A capillary HPLC (cHPLC) coupled with diode array detection (DAD) and MS method was developed for the simultaneously qualitative and quantitative determination of nine components, namely vanillic acid, calycosin-7-O-β-D-glucoside, (6αR,11αR)-9,10-dimethoxypterocarpan-3-O-β-D-glucoside, ononin, calycosin, (3R)-2′-hydroxy-3′,4′-dimethoxyisoflavan-7-O-β-D-glucoside, isoliquiritigenin, formononetin, (3R)-8,2′-dihydroxy-7,4′-dimethoxyisoflavan, in Radix Hedysari (Hongqi) and Radix Astragali (Huangqi). Simultaneous separation of these nine compounds was achieved on a Zorbax C18 microcolumn (5 μm, 150×0.3 mm). The mobile phase consisted of (A) 0.3% aqueous formic acid and (B) ACN with a gradient elution. The identification of nine compounds in both Hongqi and Huangqi was confirmed by TOF-MS. All calibration curves showed good linearity (R2 >0.998) within test ranges. This method showed good repeatability for the quantification of these nine components in Hongqi and Huangqi with intra- and inter-day variations of less than 1.89 and 3.13%, respectively. The validated method was successfully applied to quantify nine investigated components in eighteen samples of Hongqi and Huangqi. Hierarchical cluster analysis of 18 samples was performed using the peak area of nine analytes on cHPLC chromatograms. The result showed that Hongqi and Huangqi are significantly different, though the two species of Astragalus are very similar.
Co-reporter:Li Xia;Hai-Ling Liu, ;Jian-Liang Zhou;Lian-Wen Qi;Ling Yi ;Jun Chen
Journal of Separation Science 2008 Volume 31( Issue 18) pp:3156-3169
Publication Date(Web):
DOI:10.1002/jssc.200800327
Abstract
By fully optimizing the separation and analytical conditions, a rapid-resolution LC (RRLC, 1.8 μm particle size) coupled with TOF-MS method was developed and validated for multiple compounds analysis of traditional Chinese medicinal compound preparations (TCMCPs). As a typical example, determination of Compound Danshen preparations (CDPs, mainly comprising Salvia miltiorrhiza and Panax notoginseng), was improved on three groups of bioactive constituents (phenolic acids, diterpenoids, and saponins). LODs down to 1.2 pg, permitting very good separation of 20 analytes and 3 internal standards (ISs) in 17 min, represented an approximate five-fold reduction of time compared to conventional HPLC. Unequivocal identification of target compounds in CDPs was based on accurate mass measurement of ions, and comparison of retention performance with available standards. Quantitation was carried out using the narrow window extracted ion chromatograms (XICs) of each compound with an interference-free baseline (using a ±0.02 Da window) yielding good linearity of response (r2 >0.9940), and excellent precision of retention time (tR) both interday (RSD, 0.01–0.19%) and intraday (RSD, 0.02–1.08%). The acceptable recoveries obtained were in the range of 77.69–113.8%. The results demonstrated the robustness and applicability of RRLC-TOF-MS for multicomponents analysis in TCMCPs with diverse chemical compositions and properties.
Co-reporter:Juan Li;Hui-Jun Li;Zhi-Ming Bi;Yue Song;Yan-Jing Li
Journal of Separation Science 2007 Volume 30(Issue 6) pp:843-850
Publication Date(Web):20 MAR 2007
DOI:10.1002/jssc.200600341
An HPLC with evaporative light scattering detection (ELSD) and ESI-MS was established for the simultaneous determination of eight triterpenoids in Radix Achyranthis Bidentatae. The optimal chromatographic conditions were achieved on a Zorbax C18 column by linear gradient elution with 0.08% v/v aqueous formic acid and ACN as the mobile phase at the flow rate of 0.8 mL/min. Temperature for the detector drift tube was set at 101°C and the nitrogen flow rate was 2.8 L/min. The identities of the analytes were accomplished by comparing retention times and mass data with those of reference compounds. The validation of the method included tests of linearity, sensitivity, repeatability, recovery, and stability. All the calibration curves of the eight triterpenoids showed good linear regression (R2 >0.997) within the test ranges. The method provides desirable repeatability with overall intra- and interday variations of less than 4.9%. The obtained recoveries varied between 93.6 and 98.1% while the RSDs were below 3.9% (n = 3). The analysis results indicate that the content of investigated triterpenoids in Radix Achyranthis Bidentatae from different locations was greatly diverse, and the triterpenoids could be used as chemical markers for the discrimination of genuine and ungenuine crude drugs.
Co-reporter:Jing Dou;Lian-Wen Qi;Zhi-Ming Bi;Yue Song
Journal of Separation Science 2007 Volume 30(Issue 7) pp:992-998
Publication Date(Web):11 APR 2007
DOI:10.1002/jssc.200600434
A liquid chromatography-mass spectrometry method is presented for the quantification of C21 steroids in the roots and rhizomes of Cynanchum paniculatum. Eight C21 steroids, including five steroidal aglycones and three steroidal glycosides, were simultaneously analyzed by liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry. The extracted ion current chromatograms were extracted from the total ion current chromatogram using characteristic ions produced by target compounds for peak determination. Chromatographic separation was achieved on a C18 reversed-phase column within 60 min, using an acetonitrile/water gradient. For comparision, six C. paniculatum samples from different locations were investigated by the established method, and the results indicated that the different geographical origin significantly influenced the C21 steroid composition. The method was observed to have the necessary sensitivity, selectivity, precision, and accuracy, and to be suitable for quality control of herbal medicines and their preparations.
Co-reporter:Xiao Qin Zhou, Zhi Ming Bi, Ping Li, Dan Tang, Hai Xia Cai
Chinese Chemical Letters 2007 Volume 18(Issue 10) pp:1221-1223
Publication Date(Web):October 2007
DOI:10.1016/j.cclet.2007.08.019
A new iridoid glycoside, 6′-O-sinapoylgeniposide, was isolated from Gardenia jasminoides Ellis and its structure was elucidated on the basis of 1D and 2D NMR, HR-ESI-MS techniques.
Co-reporter:Qing Tao Yu, Ping Li, Zhi Ming Bi, Jun Luo, Xiao Dan Gao
Chinese Chemical Letters 2007 Volume 18(Issue 5) pp:554-556
Publication Date(Web):May 2007
DOI:10.1016/j.cclet.2007.03.025
Two new saponins named mongholicoside A (1) and mongholicoside B (2) were isolated from the aerial part of Astragalus membranaceus var. mongholicus. Their structures were determined by 1D and 2D NMR, ESI-MS techniques and chemical methods.
Co-reporter:Ping Li;Shi-Hui Qian;Jian-Feng Zhang;Xiao-Jie Gu;You-Bin Li
Helvetica Chimica Acta 2007 Volume 90(Issue 1) pp:72-78
Publication Date(Web):26 JAN 2007
DOI:10.1002/hlca.200790023
Three new oleanane-skeleton triterpenoid saponins, 3β,4β,16α-17-carboxy-16,24-dihydroxy-28-norolean-12-en-3-yl 4-O-β-D-xylopyranosyl-β-D-glucopyranosiduronic acid (1), (3β,4β,16α)-17-carboxy-16,24-dihydroxy-28-norolean-12-en-3-yl β-D-glucopyranosiduronic acid methyl ester (2), and (3β,4β)-24-hydroxy-16-oxo-28-norolean-12-en-3-yl 4-O-β-D-xylopyranosyl-β-D-glucopyranosiduronic acid (3), together with eight known constituents, i.e., the oleanane-type triterpenoids 4–6, and the ursane-type triterpenoids 7–11, were isolated from the spikes of Prunella vulgaris. The new structures were established by means of detailed spectroscopic analysis (IR, HR-ESI-MS, 1D- and 2D-NMR experiments). Compounds 1–3 were tested for their inhibition activity against the growth of tumor cell lines; only compound 3 displayed marginal inhibition activity.
Co-reporter:Li-Wei Wang;Hui-Jun Li;Ying-Jie Wei;Dan Tang;Ling Yi
Chromatographia 2007 Volume 66( Issue 5-6) pp:395-399
Publication Date(Web):2007 September
DOI:10.1365/s10337-007-0320-9
The objective of this research was to establish a simple, practical and efficient method for routine quantitative analysis of Erigeron breviscapus and its extract injection to control their qualities. The reversed phase high performance liquid chromatographic method was adopted to determine simultaneously the contents of two major classes of constituents namely phenolic acids and flavonoids, which were usually ignored in previous studies of E. breviscapus. Under the optimum conditions, three flavonoids including scutellarin, scutellarein and apigenin and four phenolic acids including caffeic acid, chlorogenic acid, 3,4-O-dicaffeoylquinic acid and 3,5-O-dicaffeoylquinic acid were successfully separated on a Zorbax SB-C18 column (250 × 4.6 mm I.D., 5.0 μm particle size) at 25 °C. Of the three flavonoids, scutellatin is a flavone glucuronide. The mobile phase was a mixture of acetonitrile and 1.0% (v/v) aqueous acetic acid employing gradient elution at a flow rate of 1.0 mL min−1 and the detection wavelength was set at 330 nm. Regression equations revealed good linear relationship between the peak areas of the analytes and their concentrations (r2> 0.9990). The relative standard deviations of retention time and peak area were less than 0.33 and 1.45%. The intra- and inter-day precisions as determined from sample solutions were below 1.66 and 2.35%. And the recoveries ranged from 96.5 to 101.8%. The proposed method has been successfully applied to the simultaneous quantification of two major classes of constituents in E. breviscapus and its extract injection for the first time.
Co-reporter:Duo Wang;Yue Song;Yun-Yun Bian;Jia Guan;Song-Lin Li
Journal of Separation Science 2006 Volume 29(Issue 13) pp:2012-2022
Publication Date(Web):28 JUL 2006
DOI:10.1002/jssc.200500486
A method has been developed for the qualitative and quantitative analysis of pharmacologically active astragalosides isolated from several species of the genus Astragalus by high performance liquid chromatography coupled with electrospray ionization time-of-flight mass spectrometry. Seven astragalosides in Radix Astragali and their commercial pharmaceutical preparations were analyzed using the developed method. The extracted ion current chromatograms were obtained from the total ion current chromatogram using the m/z of [M+Na]+ ions produced by target compounds for peak determination. The limits of detection and limits of quantification were in the range of 0.10–0.22 ng and 0.22–0.52 ng in full scan mode, respectively. All calibration curves showed good linear regression (r2 ⪈0.9965) within the test range. The overall intra- and inter-day precision was less than 2.86% for peak area and the accuracy was higher than 92.9% on using ginsenoside I as internal standard. The assay was successfully utilized to analyze the major biologically active astragalosides in six samples of Astragalus membranaceus (Fisch.) Bge var. mongholicus (Bge.) Hsiao. and eight commercial preparations. The overall results demonstrate that this method is simple, selective, and suitable for the quality control of Chinese medicine and their preparation in the low nanogram range.
Co-reporter:Hui-juan Liu, Hao-bin Hu, Chu Chu, Qin Li, Ping Li
Acta Pharmaceutica Sinica B (October 2011) Volume 1(Issue 3) pp:189-195
Publication Date(Web):October 2011
DOI:10.1016/j.apsb.2011.06.013
Co-reporter:Zhishen Xie, Xiaomeng Wan, Lingjun Zhong, Hua Yang, Ping Li, Xiaojun Xu
Journal of Functional Foods (April 2017) Volume 31() pp:
Publication Date(Web):April 2017
DOI:10.1016/j.jff.2017.01.040
•CA is a novel SREBP inhibitor.•CA stimulates the degradation of the mature form of SREBPs.•CA promotes the degradation of n-SREBPs via the 26S proteasome.•CA alleviated dyslipidemia induced by WD.•CA improved the insulin resistance induced by WD.Carnosic acid (CA), from rosemary, a food additive used for long history in western countries, reduces triglyceride, cholesterol and glucose levels. However, the detailed mechanisms contributing to these effects remain unclear. Here we investigate the potential mechanism of how CA regulates lipid metabolism. Using reporter cell lines, we found CA inhibited SREBPs activity, using radioactive isotope labeling, we found CA impeded de novo lipid synthesis, using gene silencing and inhibitors, we investigated the possible mechanism of how CA affects SREBPs stability. In hepatocytes, CA reduces the nuclear abundance of SREBPs and downregulates their target genes, thereby inhibiting the de novo biosynthesis of both fatty acids and cholesterol. Furthermore, in western type diet-induced obese mice, CA alleviates hyperlipidemia, fatty liver and insulin resistance by suppressing SREBPs target gene expression in the liver. Our results indicate that CA promotes the degradation of mature form of SREBPs via the 26S proteasome. To our knowledge, this study is the first to describe a small molecule that strongly reduces mature SREBPs.
Co-reporter:Hui-Peng Song, Hong Wang, Xiaoai Zhao, Ling He, Huailing Zhong, Si-Qi Wu, Ping Li, Hua Yang
Journal of Hazardous Materials (5 July 2017) Volume 333() pp:265-274
Publication Date(Web):5 July 2017
DOI:10.1016/j.jhazmat.2017.03.025
•Dynamic mass redistribution (DMR) profiling was proposed to characterize toxic herbs.•Qualitative DMR profiling has obvious advantages over the classic method.•Quantitative DMR profiling is powerful in discovery of toxic marker compounds.•The proposed method is high-throughput, cost-saving and easily operated.•DMR profiling is universal for discovery of most receptor-targeted toxicants.Natural products are becoming increasingly popular in multiple fields involving medicines, foods and beverages. However, due to the frequent occurrence of poisoning incidents, their toxicity and safety have caused a serious concern. Here we report a method of biosensor-based two-phase pharmacological profiling (BTPP) for discovery, monitor and control of receptor-targeted natural products. BTPP uses a resonant waveguide grating biosensor for label-free and non-invasive detection of intracellular dynamic mass redistribution (DMR), a phenomenon caused by protein relocalization after receptors receiving stimulation from toxicants. The method can not only facilitate the identification of hazardous materials but also quantify their bioactivity by EC50. As a proof of concept, the method was successfully applied to recognize Daturae Flos (DF), an herb that can antagonize muscarinic acetylcholine M2 receptor and further cause poisoning, from other easily confused species. BTPP combined with high performance liquid chromatography revealed that scopolamine and hyoscyamine in DF were the key marker compounds. Moreover, the method accurately picked out 2 M2 receptor antagonists from 25 natural compounds, displaying its potential in high-throughput screening. This study provides a systematic illustration about the establishment, applicability and advantages of BTPP, which contributes to the safety assessment of natural products in related fields.Download high-res image (151KB)Download full-size image
Co-reporter:Yong Fan, Yong Li, Yan Chen, Yi-Jing Zhao, ... Lian-Wen Qi
Journal of the American College of Cardiology (20 September 2016) Volume 68(Issue 12) pp:1281-1293
Publication Date(Web):20 September 2016
DOI:10.1016/j.jacc.2016.06.044
BackgroundPathogenesis and diagnostic biomarkers for diseases can be discovered by metabolomic profiling of human fluids. If the various types of coronary artery disease (CAD) can be accurately characterized by metabolomics, effective treatment may be targeted without using unnecessary therapies and resources.ObjectivesThe authors studied disturbed metabolic pathways to assess the diagnostic value of metabolomics-based biomarkers in different types of CAD.MethodsA cohort of 2,324 patients from 4 independent centers was studied. Patients underwent coronary angiography for suspected CAD. Groups were divided as follows: normal coronary artery (NCA), nonobstructive coronary atherosclerosis (NOCA), stable angina (SA), unstable angina (UA), and acute myocardial infarction (AMI). Plasma metabolomic profiles were determined by liquid chromatography–quadrupole time-of-flight mass spectrometry and were analyzed by multivariate statistics.ResultsWe made 12 cross-comparisons to and within CAD to characterize metabolic disturbances. We focused on comparisons of NOCA versus NCA, SA versus NOCA, UA versus SA, and AMI versus UA. Other comparisons were made, including SA versus NCA, UA versus NCA, AMI versus NCA, UA versus NOCA, AMI versus NOCA, AMI versus SA, significant CAD (SA/UA/AMI) versus nonsignificant CAD (NCA/NOCA), and acute coronary syndrome (UA/AMI) versus SA. A total of 89 differential metabolites were identified. The altered metabolic pathways included reduced phospholipid catabolism, increased amino acid metabolism, increased short-chain acylcarnitines, decrease in tricarboxylic acid cycle, and less biosynthesis of primary bile acid. For differential diagnosis, 12 panels of specific metabolomics-based biomarkers provided areas under the curve of 0.938 to 0.996 in the discovery phase (n = 1,086), predictive values of 89.2% to 96.0% in the test phase (n = 933), and 85.3% to 96.4% in the 3-center external sets (n = 305).ConclusionsPlasma metabolomics are powerful for characterizing metabolic disturbances. Differences in small-molecule metabolites may reflect underlying CAD and serve as biomarkers for CAD progression.
Co-reporter:Mu Zhang, Yanyan Wang, Fei Qian, Ping Li, Xiaojun Xu
Biochemical and Biophysical Research Communications (2 December 2016) Volume 481(Issues 1–2) pp:
Publication Date(Web):2 December 2016
DOI:10.1016/j.bbrc.2016.11.016
•Hypericin suppresses oligomeric amyloid β42-induced pro-inflammatory cytokines expression in BV2 cells.•Hypericin ameliorates learning and memory deficiency in an amyloid β injection mouse model of Alzheimer's disease.•Hypericin suppresses inflammation response in microglia through suppressing MKL1.Amyloid β (Aβ) provokes severe inflammation response in the central nervous system, which is a key risk factor for the progression of Alzheimer's disease (AD). Anti-inflammation medications shed light on treating AD. In this study, we found hypericin is a potent anti-AD constituent through anti-inflammation. Pretreatment with hypericin (5 μM and 15 μM) significantly suppresses oligomeric Aβ42 (oAβ42)-induced expression of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor α (TNF α) and inducible nitric oxide synthase (iNOS) and production of NO in microglia without cytotoxicity. We further found that hypericin ameliorates inflammatory response by suppressing MKL1, which is the essential cofactor of p65 during the transcription process. In an Aβ injection AD mouse model, animals orally administrated hypericin (50 mg/kg) for seven days significantly decreased pro-inflammatory cytokines expression and NO production in hippocampus, meanwhile, hypericin improved oAβ42-induced learning and memory impairment in mice in the Morris water maze test. Therefore, hypericin could be considered as a potential candidate for treating AD.
Co-reporter:Meijuan Xu, Jungang Yin, Liyan Xie, Jun Zhang, Chong Zou, Jiandong Zou, Fang Liu, Wenzheng Ju, Ping Li
Phytomedicine (15 September 2013) Volume 20(Issue 12) pp:1105-1111
Publication Date(Web):15 September 2013
DOI:10.1016/j.phymed.2013.05.004
Total astragalosides (TA) are the principal active constituents isolated from Radix Astragali, which has been extensively used in the traditional Chinese medicine for hundreds of years. However, few detailed pharmacokinetic studies about TA or its main component in human have been done to date. The aim of this study was to investigate the pharmacokinetic (PK) characteristics of astragaloside IV (AGS-IV), the primary ingredient of TA, and tolerance of TA after single- and multi-intravenous infusion of astragalosides injection (AI) in healthy Chinese volunteers. A LC–MS/MS assay was developed for AGS-IV determination in human plasma and urine, and the PK parameters were estimated using non-compartmental methods. The mean maximum plasma concentration (Cmax) values of AGS-IV were 2.12, 3.59, 3.71 and 5.17 μg ml−1 after single doses of 200, 300, 400 and 500 ml of AI, respectively. The corresponding mean values of area under the plasma concentration (AUC0−∞) were 4.38, 9.75, 13.59 and 18.22 μg h ml−1, respectively, and the mean values of elimination half-life (t1/2) were 2.14, 2.59, 2.62 and 2.69 h, respectively. In the repeated dose study, no significant difference was observed between the PK parameters, peak time (Tmax), t1/2 and AUC, of day 1 and day 7. Cumulative urinary excretion of AGS-IV was 3.91% within 24 h after administration of 500 ml AI. AI was safe and well tolerated, and the adverse events, such as raised total bilirubin and rash, were mild and resolved spontaneously. In summary, the pharmacokinetic properties of AGS-IV are based on linear pharmacokinetics over the doses ranging from 200 to 500 ml of AI. No accumulation of AGS-IV was observed after repeated administration of AI once daily. AI was safe and well tolerated in this study, although cases of transient adverse events were observed.Download high-res image (145KB)Download full-size image
Co-reporter:Liu Zhang, Hui Zhang, Xueyan Li, Bingjie Jia, Yuyu Yang, Ping Zhou, Ping Li, Jun Chen
Phytomedicine (15 December 2016) Volume 23(Issue 14) pp:1806-1813
Publication Date(Web):15 December 2016
DOI:10.1016/j.phymed.2016.11.003
BackgroundOxidized low-density lipoprotein (ox-LDL) is an underlying cause of endothelial dysfunction, which is an early event in the pathogenesis of atherosclerosis. In our previous study, we established an ARE-driven luciferase reporter system and screened out several potential Nrf2 activators from Salvia miltiorrhiza Bunge.PurposeSince miltirone showed the most potent ARE-driven luciferase activity, the aim of this study was to test the protective role of miltirone against oxidative stress in endothelial cell and to investigate the underlying mechanistic signaling pathways.Study Design/MethodIn the present study, miltirone increased the expression of nuclear translocation and transcriptional activities of NF-E2-related factor 2 (Nrf2), which led to augmented expression of antioxidant-response element (ARE)-dependent heme oxygenase-1 (HO-1) and NAD(P)H-quinone oxidoreductase 1 (NQO1). Inhibition of Nrf2/HO-1 by RNA interference abolished miltirone-induced cytoprotective effects against ox-LDL, which suggested that Nrf2 and the downstream expression of HO-1 are required for the functional effects of miltirone. Ox-LDL-stimulated mitogen-activated protein kinase activation, ROS production, and miltirone dramatically inhibited synthesis of ROS, as well as decreased SOD and glutathione S-transferase (GST) in human EA.hy926 endothelial cells.ResultsMiltirone-induced Nrf2 and HO-1 expression was related to mitogen-activated protein kinase (MAPK) pathways. The activation of MAPK was partially dependent on the phosphorylation of the c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) pathways, but not P38 MAPK signaling. However, miltirone-induced Nrf2/HO-1 expression can only be effectively blocked by JNK inhibitor SP600125.ConclusionOur findings reveal that miltirone exerts protective functions on endothelial cells in response to ox-LDL-induced oxidative stress, and does so via Nrf2/HO-1, which provides novel insights into the antioxidant capacity of miltirone.Download high-res image (286KB)Download full-size image
Co-reporter:Meijuan Xu, Haiping Hao, Lifeng Jiang, Fang Long, Yidan Wei, Hui Ji, Bingting Sun, Ying Peng, Guangji Wang, Wenzheng Ju, Ping Li
Phytomedicine (15 April 2015) Volume 22(Issue 4) pp:444-451
Publication Date(Web):15 April 2015
DOI:10.1016/j.phymed.2015.02.001
Background: Soluble epoxide hydrolase (sEH) has been demonstrated to be a key enzyme involved in the pathologic development of several cardiovascular diseases and inflammation, and inhibition of sEH is therefore very helpful or crucial for the treatment of ischemia-reperfusion injury, cardiac hypertrophy, hypertension and inflammation. Danshen, the dried root of Salvia miltiorrhiza (Fam. Labiatae), has been used for the treatment of cardiovascular and cerebrovascular diseases in China and other countries for hundreds of years. Recent studies indicated that Danshen and its preparations also have potential for the management of inflammation. However, little information is available about the possibility of Danshen and its components on sEH inhibition.Purpose and methods: Danshen extracts and its constituents were tested for sEH inhibition using its physiological substrate, 8,9-EET, based on a LC–MS/MS assay in this study.Results: Among the tested 15 compounds, tanshinone IIA and cryptotanshinone were found to be the potent (Ki = 0.87 μM) and medium (Ki = 6.7 μM) mixed-type inhibitors of sEH, respectively. Salvianolic acid C (Ki = 8.6 μM) was proved to be a moderate noncompetitive sEH inhibitor. In consistent with the inhibition results of the pure compounds, the 75% ethanol extract of Danshen (EE, IC50 = 86.5 μg/ml) which contained more tanshinone IIA and cryptotanshinone exhibited more potent inhibition on sEH than the water extract (WE, IC50 > 200 μg/ml) or 1 M NaHCO3 (BE, IC50 > 200 μg/ml) extract.Conclusion: These data indicated that using the ethanol fraction of Danshen and increasing the amounts of tanshinone IIA, cryptotanshinone and salvianolic acid C, especially the contents of tanshinone IIA in Danshen extract or preparations to enhance the inhibitory effects on sEH might be efficient ways to improve its cardiovascular protective and anti-inflammatory effects, and that herbal medicines could be an untapped reservoir for sEH-inhibition agents and developing sEH inhibitors from the cardiovascular protective and anti-inflammatory herbs is a promising approach.Download high-res image (178KB)Download full-size image
Co-reporter:Hui-Peng Song, Jun Chen, Jia-Ying Hong, Haiping Hao, Lian-Wen Qi, Jun Lu, Yu Fu, Bin Wu, Hua Yang and Ping Li
Chemical Communications 2015 - vol. 51(Issue 8) pp:NaN1497-1497
Publication Date(Web):2014/12/02
DOI:10.1039/C4CC08728C
A novel strategy of ultrafiltration LC-MS and in silico molecular docking was proposed to discover high-quality enzyme inhibitors from herbal medicines. Using this strategy, two compounds were predicted and finally demonstrated as potent xanthine oxidase inhibitors, whose in vitro IC50 values were lower than that of a positive control allopurinol.
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![2-METHYL-2-PROPANYL 4-{[(6-CHLORO-3-PYRIDAZINYL)AMINO]METHYL}-1-P<WBR />IPERIDINECARBOXYLATE](http://img.cochemist.com/ccimg/37000/36948-76-2.png)
![2-METHYL-2-PROPANYL 4-{[(6-CHLORO-3-PYRIDAZINYL)AMINO]METHYL}-1-P<WBR />IPERIDINECARBOXYLATE](http://img.cochemist.com/ccimg/37000/36948-76-2_b.png)
![[1S-(1α,4aα,6α,7α,7aα)]-1-(β-D-glucopyranosyloxy)-1,4a,5,6,7,7a-hexahydro-6-hydroxy-7-methylcyclopenta[c]pyran-4-carboxylic acid](http://img.cochemist.com/ccimg/22300/22255-40-9.png)
![[1S-(1α,4aα,6α,7α,7aα)]-1-(β-D-glucopyranosyloxy)-1,4a,5,6,7,7a-hexahydro-6-hydroxy-7-methylcyclopenta[c]pyran-4-carboxylic acid](http://img.cochemist.com/ccimg/22300/22255-40-9_b.png)
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![Didymin with HPLC [14259-47-3]](http://img.cochemist.com/ccimg/14300/14259-47-3_b.png)
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![[PHENYL(PHENYLSULFANYL)PHOSPHORYL]BENZENE](http://img.cochemist.com/ccimg/5600/5510-78-1.png)
![[PHENYL(PHENYLSULFANYL)PHOSPHORYL]BENZENE](http://img.cochemist.com/ccimg/5600/5510-78-1_b.png)
![(e)-1-(2,4-dihydroxyphenyl)-3-[4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]prop-2-en-1-one](http://img.cochemist.com/ccimg/5100/5041-81-6.png)
![(e)-1-(2,4-dihydroxyphenyl)-3-[4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]prop-2-en-1-one](http://img.cochemist.com/ccimg/5100/5041-81-6_b.png)
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![Bicyclo[3.1.0]hex-2-ene, 2-methyl-5-(1-methylethyl)-](http://img.cochemist.com/ccimg/2900/2867-05-2.png)
![Bicyclo[3.1.0]hex-2-ene, 2-methyl-5-(1-methylethyl)-](http://img.cochemist.com/ccimg/2900/2867-05-2_b.png)
![Isoquinoline,1-[(3,4-dimethoxyphenyl)methyl]-1,2,3,4-tetrahydro-6,7-dimethoxy-2-methyl-,(1S)-](http://img.cochemist.com/ccimg/2700/2688-77-9.png)
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![5-hydroxy-3-(4-hydroxyphenyl)-6-methoxy-7-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/700/611-40-5_b.png)
![(2s)-7-hydroxy-2-[4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]-2,3-dihydrochromen-4-one](http://img.cochemist.com/ccimg/600/551-15-5.png)
![(2s)-7-hydroxy-2-[4-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]-2,3-dihydrochromen-4-one](http://img.cochemist.com/ccimg/600/551-15-5_b.png)
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![4H-1-Benzopyran-4-one,5,7-dihydroxy-8-[5-(5-hydroxy-7-methoxy-4-oxo-4H-1-benzopyran-2-yl)-2-methoxyphenyl]-2-(4-methoxyphenyl)-](http://img.cochemist.com/ccimg/600/521-34-6.png)
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![2-(3,4-dihydroxyphenyl)-3,5-dihydroxy-7-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/500/491-50-9_b.png)
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![9-(3-methylbut-2-enyloxy)-7H-furo[3,2-g]chromen-7-one](http://img.cochemist.com/ccimg/500/482-44-0_b.png)
![3-[(2S,3R,4R,5R,6S)-4,5-DIHYDROXY-6-METHYL-3-[(2S,3R,4S,5S,6R)-3,4,5-TRIHYDROXY-6-(HYDROXYMETHYL)OXAN-2-YL]OXYOXAN-2-YL]OXY-5,7-DIHYDROXY-2-(4-HYDROXYPHENYL)CHROMEN-4-ONE](/data/chemimg/513000/142451-65-8.png)
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![Phenanthro[1,2-b]furan-10,11-dione, 1-(hydroxymethyl)-6-methyl-](/data/chemimg/2084200/76829-01-1_b.png)
![5,7-dihydroxy-2-(4-hydroxyphenyl)-6-[(2s,3r,4r,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]-8-[(2s,3r,4s,5s)-3,4,5-trihydroxyoxan-2-yl]chromen-4-one](http://img.cochemist.com/ccimg/52000/51938-32-0.png)
![5,7-dihydroxy-2-(4-hydroxyphenyl)-6-[(2s,3r,4r,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]-8-[(2s,3r,4s,5s)-3,4,5-trihydroxyoxan-2-yl]chromen-4-one](http://img.cochemist.com/ccimg/52000/51938-32-0_b.png)
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![[(2r,3r,4s,5r,6s)-3,4,5,6-tetrakis[(3,4,5-trihydroxybenzoyl)oxy]oxan-2-yl]methyl 3,4,5-trihydroxybenzoate](http://img.cochemist.com/ccimg/15000/14937-32-7.png)
![[(2r,3r,4s,5r,6s)-3,4,5,6-tetrakis[(3,4,5-trihydroxybenzoyl)oxy]oxan-2-yl]methyl 3,4,5-trihydroxybenzoate](http://img.cochemist.com/ccimg/15000/14937-32-7_b.png)
![(7s,13as)-3,10-dimethoxy-7-methyl-6,8,13,13a-tetrahydro-5h-isoquinolino[2,1-b]isoquinolin-7-ium-2,11-diol](http://img.cochemist.com/ccimg/6900/6873-13-8.png)
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![Bis[1,3]benzodioxolo[5,6-a:4',5'-g]quinolizinium,6,7-dihydro-](http://img.cochemist.com/ccimg/3500/3486-66-6.png)
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![3,5,9-Trioxa-4-phosphapentacosan-1-aminium,4-hydroxy-N,N,N-trimethyl-10-oxo-7-[(1-oxohexadecyl)oxy]-, inner salt, 4-oxide](http://img.cochemist.com/ccimg/2700/2644-64-6.png)
![3,5,9-Trioxa-4-phosphapentacosan-1-aminium,4-hydroxy-N,N,N-trimethyl-10-oxo-7-[(1-oxohexadecyl)oxy]-, inner salt, 4-oxide](http://img.cochemist.com/ccimg/2700/2644-64-6_b.png)
![2-Propanol, 1-[(1-methylethyl)amino]-3-(1-naphthalenyloxy)-](/data/chemimg/935900/525-66-6.png)
![2-Propanol, 1-[(1-methylethyl)amino]-3-(1-naphthalenyloxy)-](/data/chemimg/935900/525-66-6_b.png)
![5-hydroxy-2-(4-methoxyphenyl)-7-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/500/480-36-4.png)
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![(1s,3r,4r,5r)-3,4-bis[[(e)-3-(3,4-dihydroxyphenyl)prop-2-enoyl]oxy]-1,5-dihydroxycyclohexane-1-carboxylic Acid](http://img.cochemist.com/ccimg/14600/14534-61-3_b.png)
![(5alpha,8xi,9xi,12alpha,13xi,14beta)-12-hydroxy-17-[(2S)-2-hydroxy-6-methylhept-5-en-2-yl]-4,4,5,10,14-pentamethylgonan-3-yl 2-O-(6-O-acetyl-beta-D-glucopyranosyl)-beta-D-glucopyranoside](http://img.cochemist.com/ccimg/194900/194861-70-6.png)
![(5alpha,8xi,9xi,12alpha,13xi,14beta)-12-hydroxy-17-[(2S)-2-hydroxy-6-methylhept-5-en-2-yl]-4,4,5,10,14-pentamethylgonan-3-yl 2-O-(6-O-acetyl-beta-D-glucopyranosyl)-beta-D-glucopyranoside](http://img.cochemist.com/ccimg/194900/194861-70-6_b.png)
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![(2r,3r)-2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-[(2s,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxy-2,3-dihydrochromen-4-one](http://img.cochemist.com/ccimg/29900/29838-67-3_b.png)
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![(S)-6-(Hydroxymethyl)-1,6-dimethyl-6,7,8,9-tetrahydrophenanthro[1,2-b]furan-10,11-dione](http://img.cochemist.com/ccimg/17400/17397-93-2.png)
![(S)-6-(Hydroxymethyl)-1,6-dimethyl-6,7,8,9-tetrahydrophenanthro[1,2-b]furan-10,11-dione](http://img.cochemist.com/ccimg/17400/17397-93-2_b.png)
![[(2r,3r,4r,5r,6r)-6-[2-(3,4-dihydroxyphenyl)ethoxy]-5-hydroxy-2-[[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]-4-[(2s,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-3-yl] (e)-3-(3,4-dihydroxyphenyl)prop-2-enoate](http://img.cochemist.com/ccimg/82900/82854-37-3.png)
![[(2r,3r,4r,5r,6r)-6-[2-(3,4-dihydroxyphenyl)ethoxy]-5-hydroxy-2-[[(2r,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxymethyl]-4-[(2s,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-3-yl] (e)-3-(3,4-dihydroxyphenyl)prop-2-enoate](http://img.cochemist.com/ccimg/82900/82854-37-3_b.png)
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![Dimethyl 3-oxo-3H-pyrrolo[2,1,5-de]quinolizine-1,2-dicarboxylate](http://img.cochemist.com/ccimg/94600/94596-27-7.png)
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![(4AR,6AR,6AS,6BR,8AR,9R,10S,12AR,14BS)-10-[(2S,4S,5S)-3-[(2S,3R,4R,5S,6S)-3,5-DIHYDROXY-6-METHYL-4-[(2S,3R,4S,5S,6R)-3,4,5-TRIHYDROXY-6-(HYDROXYMETHYL)OXAN-2-YL]OXYOXAN-2-YL]OXY-4,5-DIHYDROXYOXAN-2-YL]OXY-9-(HYDROXYMETHYL)-2,2,6A,6B,9,12A-HEXAMETHYL-1,3,4](http://img.cochemist.com/ccimg/128800/128730-82-5.png)
![(4AR,6AR,6AS,6BR,8AR,9R,10S,12AR,14BS)-10-[(2S,4S,5S)-3-[(2S,3R,4R,5S,6S)-3,5-DIHYDROXY-6-METHYL-4-[(2S,3R,4S,5S,6R)-3,4,5-TRIHYDROXY-6-(HYDROXYMETHYL)OXAN-2-YL]OXYOXAN-2-YL]OXY-4,5-DIHYDROXYOXAN-2-YL]OXY-9-(HYDROXYMETHYL)-2,2,6A,6B,9,12A-HEXAMETHYL-1,3,4](http://img.cochemist.com/ccimg/128800/128730-82-5_b.png)
![dimethyl (2S,3R,2'S,3'R,4'R)-4,4'-[(2Z)-4-oxobut-2-ene-1,3-diyl]bis[3-ethenyl-2-(beta-D-glucopyranosyloxy)-3,4-dihydro-2H-pyran-5-carboxylate]](http://img.cochemist.com/ccimg/82500/82474-97-3.png)
![dimethyl (2S,3R,2'S,3'R,4'R)-4,4'-[(2Z)-4-oxobut-2-ene-1,3-diyl]bis[3-ethenyl-2-(beta-D-glucopyranosyloxy)-3,4-dihydro-2H-pyran-5-carboxylate]](http://img.cochemist.com/ccimg/82500/82474-97-3_b.png)
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![5,7-dihydroxy-2-(4-hydroxyphenyl)-3-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/500/480-10-4.png)
![5,7-dihydroxy-2-(4-hydroxyphenyl)-3-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/500/480-10-4_b.png)
![5,7-dihydroxy-2-(4-hydroxyphenyl)-3-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/17700/17650-84-9.png)
![5,7-dihydroxy-2-(4-hydroxyphenyl)-3-[(2s,3r,4s,5s,6r)-3,4,5-trihydroxy-6-[[(2r,3r,4r,5r,6s)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxymethyl]oxan-2-yl]oxychromen-4-one](http://img.cochemist.com/ccimg/17700/17650-84-9_b.png)
![Benzenepropanoic acid, a-[[(2E)-3-[2-(carboxymethyl)-3,4-dihydroxyphenyl]-1-oxo-2-propen-1-yl]oxy]-3,4-dihydroxy-,(aR)-](http://img.cochemist.com/ccimg/143000/142998-47-8.png)
![Benzenepropanoic acid, a-[[(2E)-3-[2-(carboxymethyl)-3,4-dihydroxyphenyl]-1-oxo-2-propen-1-yl]oxy]-3,4-dihydroxy-,(aR)-](http://img.cochemist.com/ccimg/143000/142998-47-8_b.png)
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![4H-1-Benzopyran-4-one,7-[[2-O-(6-deoxy-a-L-mannopyranosyl)-b-D-glucopyranosyl]oxy]-2-(3,4-dihydroxyphenyl)-5-hydroxy-](http://img.cochemist.com/ccimg/25700/25694-72-8.png)
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![2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3-[(2R,3S,4R,5S,6S)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl]oxy-chromen-4-one](http://img.cochemist.com/ccimg/500/482-35-9.png)
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![Phenanthro[1,2-b]furan-10,11-dione, 1,2-dihydro-1,6-dimethyl-, (1R)-](/data/chemimg/1469400/87205-99-0.png)
![Phenanthro[1,2-b]furan-10,11-dione, 1,2-dihydro-1,6-dimethyl-, (1R)-](/data/chemimg/1469400/87205-99-0_b.png)
![(2s,3s)-4-[(e)-3-[(1r)-1-carboxy-2-(3,4-dihydroxyphenyl)ethoxy]-3-oxoprop-1-enyl]-2-(3,4-dihydroxyphenyl)-7-hydroxy-2,3-dihydro-1-benzofuran-3-carboxylic Acid](http://img.cochemist.com/ccimg/28900/28831-65-4.png)
![(2s,3s)-4-[(e)-3-[(1r)-1-carboxy-2-(3,4-dihydroxyphenyl)ethoxy]-3-oxoprop-1-enyl]-2-(3,4-dihydroxyphenyl)-7-hydroxy-2,3-dihydro-1-benzofuran-3-carboxylic Acid](http://img.cochemist.com/ccimg/28900/28831-65-4_b.png)
