Co-reporter:Mengge Zhou;Xiaoyao Ma;Jixue Sun;Guoyu Ding
Analytical and Bioanalytical Chemistry 2017 Volume 409( Issue 4) pp:1145-1157
Publication Date(Web):29 October 2016
DOI:10.1007/s00216-016-0043-6
Tropane alkaloids (TAs), rich in the plant of Physochlaina infundibularis Kuang, which is named Huashanshen (HSS) in China, showed good effects on types of spasms. However, no data were collected to explore the relationship between the specificity for muscarinic receptor subtypes and the structures of these TAs. To address this issue, an extracted ion chromatogram (EIC) strategy using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UPLC-Q/TOF-MS) based on the fragmentation behavior of the TA standards was established to rapidly capture the varied TAs from HSS. Based on the provided structural information of diagnostic ions or neutral loss, 29 TAs were efficiently profiled, especially some trace ingredients. In additional, via virtual validation combined with molecular dynamic simulation, approximately a dozen alkaloids were found with high selectivity for muscarinic receptors. In additional, N-acetyl convolicine was chosen for selectivity evaluation of M2 or M3 receptors through the use of a dual-luciferase reporter assay system at the cellular level and an ACh-induced constricted strip test in vitro. After summarizing the active fragments and the structure-activity relationship (SAR) information, a new modified TA that takes advantage of both the high affinity and high selectivity for M3 receptors was proposed and evaluated successfully. This study provided an effective approach for the discovery and design of natural products based on highly selective drugs by UPLC-Q/TOF-MS coupled with virtual calculation and biological evaluation.
Co-reporter:Wenjuan Liu;Guangcui Chu;Nianwei Chang;Xiaoyao Ma;Min Jiang
RSC Advances (2011-Present) 2017 vol. 7(Issue 64) pp:40418-40426
Publication Date(Web):2017/08/16
DOI:10.1039/C7RA06302D
Phillygenin (Phi) is one of the main chemical constituents of the fruit of Forsythia suspensa (Thunb.) Vahl. It has various bioactivities, including anti-inflammatory, anti-obesity and antipyretic activities. However, its exact targets and molecular mechanism are still poorly understood. Bioinformatics tools were used to explore the potential targets of Phi, and 8 predicted targets, 4 primary pathways (MAPK, PI3K-Akt, T-cell receptor and m-TOR signaling pathway) related to the inflammatory response, and Akt as an important node was mentioned. Moreover, a Phi alkylated molecular probe was synthesized and used to capture the target proteins Akt. Then Akt and its downstream signaling pathway were verified by molecular docking, intracellular enzyme activity evaluation, and accurate pathway analysis. The results indicated that Phi targets an allosteric inhibit pocket on Akt; reduces Akt phosphorylation; alleviates multiple inflammatory-associated downstream signal transduction pathways, including IKKα/β and NF-κB; and influences glucose metabolic parameters associated with the downstream GSK3β protein and glucose uptake. The results suggest that Phi could reduce inflammatory responses and influence glucose metabolic parameters by inhibiting Akt phosphorylation. Moreover, these findings suggest a potential application for Phi in respiratory and metabolic diseases therapy.
Co-reporter:Qingxin Cui;Yuanyuan Hou;Yanan Wang;Xu Li;Yang Liu
Journal of Nanobiotechnology 2017 Volume 15( Issue 1) pp:
Publication Date(Web):2017 December
DOI:10.1186/s12951-017-0263-8
Tracking targets of natural products is one of the most challenging issues in fields ranging from pharmacognosy to biomedicine. It is widely recognized that the biocompatible nanoparticle (NP) could function as a “key” that opens the target “lock”.We report a functionalized poly-lysine NP technique that can monitor the target protein of arctigenin (ATG) in vivo non-invasively. The NPs were synthesized, and their morphologies and surface chemical properties were characterized by transmission electron microscopy (TEM), laser particle size analysis and atomic force microscopy (AFM). In addition, we studied the localization of ATG at the level of the cell and the whole animal (zebrafish and mice). We demonstrated that fluorescent NPs could be ideal carriers in the development of a feasible method for target identification. The distributions of the target proteins were found to be consistent with the pharmacological action of ATG at the cellular and whole-organism levels.The results indicated that functionalized poly-lysine NPs could be valuable in the multimodal imaging of arctigenin.
Co-reporter:Guoyu Ding;Yanshuai Wang;Aina Liu;Yuanyuan Hou;Tiejun Zhang;Changxiao Liu
RSC Advances (2011-Present) 2017 vol. 7(Issue 36) pp:22034-22044
Publication Date(Web):2017/04/19
DOI:10.1039/C6RA28152D
Because of the poor discovery rate of relatively effective components and elusive component-effect correlation, formulating a quality control system for Chinese herbal medicines (CHMs) has been a great challenge for quality management. In this paper, the concept of the quality marker (Q-marker) was used, and a set of integrated strategies to improve the chemical markers of the Q-markers was introduced. Two often confused CHMs, Lonicera japonica flos (LJF) and Lonicera flos (LF), which are of the same genus but different species, are illustrated to quickly evaluate their potency. Ultra-performance liquid chromatography-quadrupole/time-of-flight (UPLC/Q-TOF) with partial least squares-discriminant analysis (PLS-DA) was used to screen the chemical markers for their herbal origin identification; then, a bioactive-guided evaluation method was performed to detect the Q-markers. As a result, four NF-κB inhibitors were proposed to be representative Q-markers for the anti-inflammatories: 3-O-caffeoylquinic acid (CA), 3,5-O-dicaffeoylquinic acid (3,5-diCQA), `iamarin, and vogeloside. After the chemometrics study, near-infrared spectroscopy (NIRS) based on the distinctive wavenumber points from the Q-markers was developed for its distinction and determination capabilities by optimum siPLS-CARS analysis (OPSC). Then, the back propagating-artificial neutral network (BP-ANN) algorithm was used to clarify the non-linear relationship between the Q-markers and their integral anti-inflammation effect. Finally, convenient and reliable fast quantitative analysis and holistic bioactivity assessment patterns were established by NIRS for the quality management of honeysuckle buds. The integrated Q-marker screen and NIRS assessment strategy was suitable for a fast quality evaluation of herbal medicines and was applied to the quality control of botanical functional foods.
Co-reporter:ZengYong Wang, Qian Wang, Man Zhang, XueYan Hu, GuoYu Ding, Min Jiang, Gang Bai
Biomedicine & Pharmacotherapy 2017 Volume 90(Volume 90) pp:
Publication Date(Web):1 June 2017
DOI:10.1016/j.biopha.2017.03.058
Cimicifuga rhizomes (CR) are used in the treatment of respiratory and cardiovascular diseases in traditional Chinese medicine, but their key effective components and mechanism of action have not yet been reported. In this study, the cardiac, antipyretic and sudorific effects of CR were evaluated using the toad heart failure in vitro model and mice fever and sweating in vivo models. Moreover, the UPLC/Q-TOF-MS-integrated β2-AR luciferase reporter gene assay system was used to screen the bioactive ingredients from CR extract, and the activity of this ingredient were verified using the above-mentioned in vitro and vivo models. Our results showed that CR had anti-heart failure, antipyretic and sweating effects, which could be antagonized by propranolol. On the other hand, cimicifugamide was screened as β2-AR agonist from CR and cimicifugamide could activate β1, 2-ARs more significantly than β3-AR in β-ARs selectivity assessment. The results not only revealed the key effective components and mechanism of CR in traditional use but also supplied a characteristic complementary ingredient for quality control of CR.Download high-res image (165KB)Download full-size image
Co-reporter:Nianwei Chang, Yanmei Li, Mengge Zhou, Jie Gao, Yuanyuan Hou, Min Jiang, Gang Bai
Biomedicine & Pharmacotherapy 2017 Volume 87(Volume 87) pp:
Publication Date(Web):1 March 2017
DOI:10.1016/j.biopha.2017.01.022
BackgroundCirsium setosum (CS) is the aboveground part of Cephalanoplos segetum Kitam. Although it has been used as a hemostatic treatment for thousands of years and is still in use today, the mechanism of CS on regulating ARs is still not clear.PurposeIn this study, we aimed to clarify the mechanism of CS on regulating ARs.MethodsWe developed a simple method based on UPLC/Q-TOF MS combined adrenergic receptor dual-luciferase reporter assay systems for the rapid determination of active constituents in CS. The mechanism of tyramine, the main active component for regulating ARs, was further investigated by an in vitro norepinephrine biotransformation test and in vivo vaso activity tests.ResultsTwo phenethylamine ARs regulators (tyramine and N-methyltyramine) in CS were characterized, and it was found that tyramine could induce vasoconstriction via regulation of α1-ARs by mediating norepinephrine synthesis.ConclusionThe hemostatic effect of CS is associated with tyramine and N-methyltyramine, via regulation of α1-ARs, and the mechanism of tyramine is related to mediating norepinephrine synthesis by enzyme catalysis.Download high-res image (123KB)Download full-size image
Co-reporter:Guoyu Ding, Baiqing Li, Yanqi Han, Aina Liu, Jingru Zhang, Jiamin Peng, Min Jiang, Yuanyuan Hou, Gang Bai
Journal of Pharmaceutical and Biomedical Analysis 2016 Volume 131() pp:391-399
Publication Date(Web):30 November 2016
DOI:10.1016/j.jpba.2016.09.008
•Boxplot analysis combined with the interval LODs theory was used to screen out the Q-marker ingredients for NIR analysis.•Machine learning approach was firstly introduced to solve the nonlinear spectrum-activity relationship.•A rapid bioactivity-based evaluation system based on near infrared spectroscopy for quality control of Flos Chrysanthemi was developed.For quality control of herbal medicines or functional foods, integral activity evaluation has become more popular in recent studies. The majority of researchers focus on the relationship between chromatography/mass spectroscopy and bioactivity, but the connection with spectrum-activity is easily ignored. In this paper, the near infrared reflection spectra (NIRS) of Flos Chrysanthemi samples were collected as a representative spectrum technology, and corresponding anti-inflammation activities were utilized to illustrate the spectrum-activity study. HPLC/Q-TOF-MS identification and heat map clustering were used to select the quality markers (Q-marker) from five cultivars of Flos Chrysanthemi. Using boxplot analysis and the interval limits of detection (LODs) theory, six crucial markers, namely, chlorogenic acid, 3,5-dicaffeoylquinic acid, 1,5-dicaffeoylquinic acid, luteoloside, apigenin-7-O-β-d-glucoside, and luteolin-7-O-6-malonylglucoside were screened out. Then partial least squares regression (PLSR) calibration models combined with synergy interval partial least squares (siPLS) and 12 different spectral pretreatment methods were developed for the parameters optimization of these Q-markers in Flos Chrysanthemi powder. After comparing the relationship between Q-marker contents and anti-inflammation activity via three machine learning approaches and PLSR, back-propagation neural network (BP-ANN) displayed a more excellent non-linear fitting effect, as its R for new batches reached 0.89. These results indicated that the integrated NIRS and bioactive strategy was suitable for fast quality management in Flos Chrysanthemi, and also applied to other botanical food quality control.
Co-reporter:Guoyu Ding, Yan Nie, Yuanyuan Hou, Zenghui Liu, Aina Liu, Jiamin Peng, Min Jiang, Gang Bai
Journal of Pharmaceutical and Biomedical Analysis 2015 Volume 114() pp:462-470
Publication Date(Web):10 October 2015
DOI:10.1016/j.jpba.2015.06.029
•An integrated technique of UPLC–Q/Tof and NIRS for quality control of TCM was proposed.•Four THPECs were firstly identified as potential markers to distinguish the authenticity of the Chinese Eaglewood.•With siPLS algorithm, the optimal waveband variables of NIR were targeted on the four THPECs.•The integrated technique presented a superior classification result than NIR full spectra and HPLC fingerprints.The application of near infrared spectroscopy (NIRS) in traditional Chinese medicine (TCM) has usually been limited by its blindness to qualitative or quantitative multivariate analysis and because its chemical significance is easily ignored. Here, an integrated technology of ultra-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry (UPLC–Q/Tof) and NIRS was proposed to set up a systematic quality control of Chinese Eaglewood (CE). UPLC–Q/Tof combined with principle compound analysis (PCA) was used to identify the marker ingredients of CE. Four types of highly oxidized 5,6,7,8-tetrahydro-2-(2-phenylethyl) chromones (THPECs) were then identified as potential markers to distinguish the authenticity of the CE. Based on the marker ingredients, the definite wavenumber intervals and spectral pretreatment pattern of NIRS were selected to act as an alternative evaluation technology directed against CE powder samples. Calibration equations were developed from the contents of the four markers, as detected by high performance liquid chromatography (HPLC) and NIRS data, using synergy interval partial least squares (siPLS) algorithm with Leave-One-Out (LOO) cross-validation. Using siPLS regression, satisfactory calibration statistics were obtained for the prediction of the marker ingredients. The correlation coefficient (r) between the predicted and reference results for the test set was used as an evaluation parameter for the models (r > 0.9). Hierarchical cluster analysis and partial least squares-discriminant analysis (PLS-DA) were applied to further analyze the quantitative results from NIRS. From this systematic method, 50 CE samples were divided into superior, qualified, unqualified, and fake samples, displaying a more elaborate division than PLS-DA, which is only based on whole NIR spectra or HPLC. This tandem technique of UPLC–Q/Tof and NIRS assessment presented in this work can be used as a rapid evaluation approach for the quality control of complicated herbal medicines.
Co-reporter:Liqiang Wang, Qingxin Cui, Yuanyuan Hou, Fang Bai, Jixue Sun, Xiaofang Cao, Pi Liu, Min Jiang, Gang Bai
Journal of Chromatography A 2013 Volume 1319() pp:88-96
Publication Date(Web):6 December 2013
DOI:10.1016/j.chroma.2013.10.035
•UPLC/Q-TOF-MS and virtual docking were integrated to identify inhibitors of multiple human α-glucosidases.•The system solved the issue of trace quantities limiting the biological study.•Distinct selectivity of acarviostatins was clarified based on structural biology.•Acarviostatin I0-1 was revealed to be strong inhibitor of MGAM-N.We propose a strategy that integrates ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) and virtual docking to identify inhibitors of multiple human α-glucosidases. UPLC yielded AIB656, an acarviostatin-containing complex, which was analyzed by Q-TOF-MS to acquire structural information and was tested for inhibition of N-terminal (MGAM-N), C-terminal (MGAM-C) catalytic domain of maltase-glucoamylase, and human pancreatic α-amylase (HPA). A systematic computational study was performed to evaluate the inhibition activity for 51 identified acarviostatins with various sizes, including trace or difficult-to-prepare ingredients. We evaluated the selectivities of three α-glucosidases to acarviostatins and revealed the strong inhibition of MGAM-N by acarviostatin I0-1. The high accuracy of the dual-screening was validated using enzyme inhibition assays, and docking was suggested as a possible mechanism for the strong inhibition of MGAM-N by acarviostatin I0-1 and of MGAM-C by acarbose (acarviostatin I01). No compound in AIB656 was suitable for inhibiting all three α-glucosidases. Compared with conventional chromatographic separation and inhibitory activity detection, integrating UPLC/Q-TOF-MS identification and virtual validation was more convenient and more reliable. This strategy clearly demonstrates that MS data-based fingerprinting is a meaningful tool not only in identifying constituents in complex matrix but also in directly screening for powerful trace ingredients in natural products.
Co-reporter:Qingxin Cui, Yuanyuan Hou, Jie Hou, Pengwei Pan, Lu-Yuan Li, Gang Bai, and Guoan Luo
Biomacromolecules 2013 Volume 14(Issue 1) pp:
Publication Date(Web):December 5, 2012
DOI:10.1021/bm301477z
Functionalized alkynyl polyvinyl alcohol magnetic microspheres (PVA MMs) were developed for the specific enrichment of sialic acid-rich glycoproteins by click chemistry. The capture capability for proteins was evaluated through a novel dual-labeled bovine serum albumin (BSA) that utilizes fluorescence resonance energy transfer (FRET). The PVA MM parameters, including the size and coverage of functionalized groups, were optimized by response surface methodology. The optimal parameters obtained were 1.25–6.31 μm in size and 48.53–73.05% in coverage. Then, the optimal PVA MMs were synthesized, and the morphology and surface chemical properties were characterized by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FT-IR). To capture glycoproteins from the cell surface, a bioorthogonal chemical method was applied to metabolically label them with an azide group. The functionalized alkynyl PVA MMs showed a high specificity and strong binding capability for glycoproteins through a [3 + 2] cycloaddition reaction. The results indicated that the functionalized alkynyl PVA MMs could be applied to the enrichment of cell glycoproteins, and the merits of the MMs suggested an attractive and potential way to facilitate glycoprotein research.
Co-reporter:Yuanyuan Hou, Xuelin Cao, Linyi Dong, Liqiang Wang, Binfeng Cheng, Qian Shi, Xiaodong Luo, Gang Bai
Journal of Chromatography A 2012 Volume 1227() pp:203-209
Publication Date(Web):2 March 2012
DOI:10.1016/j.chroma.2012.01.008
Although chromatographic fingerprinting combined with chemometrics, is a rational method for the quality control of traditional Chinese medicine (TCM), chemometrics cannot fully explore the relationship between chemical information and the efficacy of the potential activity. In the present work, a cell-based β2 adrenergic receptor (β2AR) agonist functional evaluation model coupled with high-performance liquid chromatography was developed to screen the potential β2AR agonist components in the alkaloidal extract of Alstonia scholaris leaves. Using a liquid chromatography with ion trap time-of-flight mass spectrometry (LCMS-IT-TOF) system, the potential bioactive compounds in the prescription were identified and deduced based on the mass spectrometric fragmentation patterns, tandem mass spectrometry (MS/MS) data, and relevant literature. Several new β2AR agonists of indole alkaloids were successfully found, and their activities were confirmed through an in vivo relaxant test on guinea pig tracheal muscles. The developed method is rapid and reliable compared with conventional fingerprinting and showed high sensitivity and resolution for the identification of β2AR agonists in TCM prescriptions. This strategy clearly demonstrates that bioactivity-integrated fingerprinting is a powerful tool not only in screening and identifying potential lead compounds and in determining the therapeutic material basis of Chinese herbal prescriptions, but also in supplying suitable chemical markers for their quality control.Highlights► We model a bioactivity-based LCMS-IT-TOF system for β2AR agonist identification. ► Two novel indole alkaloid-type β2AR agonists were characterized from A. scholaris extract. ► Bioactivity-based LCMS-IT-TOF system has advantages over conventional methods. ► This strategy is suitable for identifying bioactive constituents in complex chemical systems.
Co-reporter:Peng-wei Pan;Qi Zhang;Jie Hou;Ze Liu;Fang Bai
Analytical and Bioanalytical Chemistry 2012 Volume 403( Issue 6) pp:1661-1670
Publication Date(Web):2012 June
DOI:10.1007/s00216-012-5989-4
Bioorthogonal chemistry refers to chemical reactions that can occur within a living system without altering native biochemical processes. Applications of this concept extend to studies on a group of biomolecules that includes glycans, proteins, and lipids. In this study, a strategy for isolating cell surface glycoproteins and based on bioorthogonal chemistry was employed to identify new cancer-related glycoproteins. A novel alkyne reagent containing one disulfide bond was synthesized for the enrichment of glycoproteins metabolized with peracetylated N-azidoacetylmannosamine, which was applied on three different cancer cell lines, and all isolated proteins were analyzed by high-performance liquid chromatography–tandem mass spectrometry. The strategy of purifying cell surface glycoproteins introduced in this article was shown to be reliable, and a total of 56 cell surface glycoproteins were identified. Neuronal cell adhesion molecule was found uniquely expressed in A549 lung adenocarcinoma, and its expression in non-small-cell lung carcinomas was detected by immunohistochemistry. Furthermore, a significant increase of neuronal cell adhesion molecule expression was identified in non-small-cell lung adenocarcinoma compared with adjacent noncancerous tissues, and could be a novel potential target and marker in cancer treatment and detection.
Co-reporter:Yuanyuan Hou, Xuelin Cao, Liqiang Wang, Binfeng Cheng, Linyi Dong, Xiaodong Luo, Gang Bai, Wenyuan Gao
Journal of Chromatography B 2012 Volume 908() pp:98-104
Publication Date(Web):1 November 2012
DOI:10.1016/j.jchromb.2012.10.004
Traditional Chinese medicines (TCMs) are generally considered complementary or alternative remedies in most Western countries. The constituents of TCMs are hard to define, and their efficacy is difficult to appraise. Thus, the development of suitable methods for evaluating the relationship between bioactivity and the chemical makeup of complex TCM mixtures remains a great challenge. In the present work, the bioactivity-integrated fingerprints of alkaloidal leaf extracts of Alstonia scholaris, a folk medicinal herb for chronic respiratory diseases, were established by ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF). This method was coupled with two dual-luciferase reporter assay systems to show nuclear factor-κB (NF-κB) inhibition and β2 adrenergic receptor (β2AR) activation. Using UPLC-Q/TOF, 18 potential candidates were identified according to unique mass spectrometric fragmentation. After in vitro biological evaluation, several indole alkaloids with anti-inflammatory and anti-asthmatic properties were found, including akuammidine, (E)-alstoscholarine, and (Z)-alstoscholarine. Compared with conventional fingerprints, the microfractionation based bioactivity-integrated fingerprints that contain both chemical and bioactivity details offer a more comprehensive understanding of the chemical makeup of plant materials. This strategy clearly demonstrated that dual bioactivity-integrated fingerprinting is a powerful tool for the improved screening and identification of potential dual-target lead compounds in complex herbal medicines.Highlights► A microfractionation bioactivity-based fingerprint of A. scholaris was established. ► This method was coupled with two dual-luciferase reporter assay systems. ► NF-κB inhibitors and β2AR agonists can be screened simultaneously in one process. ► Three dual-functional indole alkaloids with dual-target were found. ► The strategy is suitable for identifying dual-target compounds in complex systems.
Co-reporter:Jie Hou;Qi Zhang;Xia Li;Yu Tang;Mei-rong Cao;Fang Bai;Qian Shi;Cui-hong Yang;De-ling Kong
Journal of Biomedical Materials Research Part A 2011 Volume 99A( Issue 4) pp:684-689
Publication Date(Web):
DOI:10.1002/jbm.a.33187
Abstract
A novel folate fluorescent nanoconjugate was synthesized and used for detection of cancer cells overexpressing the folate receptor (FR). The folate conjugate (PCMS-NA-FA) was synthesized by conjugating folic acid (FA) and 4-ethylnyl-N-ethyl-1, 8-naphthalimide (NA) to the polychloromethylstyrene (PCMS) functionalized with azido group (PCMS-N3) through click reaction. The obtained conjugate had clear structure and could form PCMS-NA-FA nanoparticles with particle size around 86 nm in aqueous solution. Ability of PCMS-NA-FA targeting to cancerous cells was investigated by comparing the uptake of the nanoparticles by human adenocarcinoma HeLa cells and by non-FR expressing human lung carcinoma A549 cells. Specificity of the PCMS-NA-FA nanoparticles targeting on FRs was verified with cellular uptake inhibition assay, in which HeLa cells were incubated with both nanoconjugate and free FA. In addition, the specificity was also confirmed by the collocation of the immunofluorescence staining of anti-FR and the cellular uptake of the PCMS-NA-FA nanoparticles. Furthermore, the organ distribution of this folate nanoconjugate was studied on HeLa cell-bearing mice via frozen slicing, and the results showed that the folate nanoparticles were preferentially accumulated in the tumor site rather than other tissues, indicating the desired specificity for tumor targeting and imaging. All these findings suggested that this practical synthetic strategy can potentially facilitate the preparation of multifunctional imaging probe in biology and diagnosis of disease. © 2011 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2011.
Co-reporter:Hao Huang;Lanxin Ji;Shengyou Song;Jing Wang;Na Wei;Min Jiang;Guoan Luo
Phytochemical Analysis 2011 Volume 22( Issue 4) pp:330-338
Publication Date(Web):
DOI:10.1002/pca.1284
Abstract
Introduction – Xuebijing injection (XBJ) is a traditional Chinese herbal prescription widely used in the treatment of sepsis. This is the first report concerning the identification of XBJ constituents. In addition, to evaluate XBJ's quality, partial least square discrimination analysis (PLS-DA) was performed on chemical fingerprint data.
Objective – Establish an LC-MS method to identify the components in XBJ for the purpose of quality control.
Methodology – Compounds were separated by HPLC using a C18 column and gradient elution of acetonitrile–methanol (60:40, v/v) and water–acetic acid (100 : 0.5, v/v) in 80 min. HPLC equipped with diode array detector (DAD) coupled with time-of-flight (TOF) tandem mass spectrometry and HPLC electrospray ionisation (ESI) multi-stage tandem ion-trap mass spectrometry (IT-MSn) method was developed to analyse XBJ's major components. Both positive and negative ionisation modes were employed.
Results – Twenty-one compounds including amino acids, phenolic acids, flavonoid glycoside, terpene glycoside and phthalide were identified or tentatively characterised. Their retention times, UV and MS spectra were compared with those of authentic compounds or literature data. The score plot of PLS-DA clearly revealed variations among samples produced in different commercial batches.
Conclusion – The analytical method developed is highly effective for the discrimination and quality control of XBJ.
Co-reporter:Limei Ren;Xiaohong Qin;Xiaofang Cao;Lele Wang;Fang Bai
Protein & Cell 2011 Volume 2( Issue 10) pp:827-836
Publication Date(Web):2011 October
DOI:10.1007/s13238-011-1105-3
Human maltase-glucoamylase (MGAM) hydrolyzes linear alpha-1,4-linked oligosaccharide substrates, playing a crucial role in the production of glucose in the human lumen and acting as an efficient drug target for type 2 diabetes and obesity. The amino- and carboxyl-terminal portions of MGAM (MGAM-N and MGAM-C) carry out the same catalytic reaction but have different substrate specificities. In this study, we report crystal structures of MGAM-C alone at a resolution of 3.1 Å, and in complex with its inhibitor acarbose at a resolution of 2.9 Å. Structural studies, combined with biochemical analysis, revealed that a segment of 21 amino acids in the active site of MGAM-C forms additional sugar subsites (+ 2 and + 3 subsites), accounting for the preference for longer substrates of MAGM-C compared with that of MGAM-N. Moreover, we discovered that a single mutation of Trp1251 to tyrosine in MGAM-C imparts a novel catalytic ability to digest branched alpha-1,6-linked oligosaccharides. These results provide important information for understanding the substrate specificity of alphaglucosidases during the process of terminal starch digestion, and for designing more efficient drugs to control type 2 diabetes or obesity.
Co-reporter:Jing Wang;Na Wei;Hongzhi Zhao;Hao Huang;Min Jiang
Chromatographia 2010 Volume 72( Issue 5-6) pp:431-440
Publication Date(Web):2010 September
DOI:10.1365/s10337-010-1677-8
High-performance liquid chromatography coupled with diode-array detection and electrospray ionization quadrupole-time of flight mass spectrometry (ESI–Q–TOF MS) has been used for global chemome analysis of Qishenyiqi dropping pills. Under gradient conditions, two sub-chemomes extracted from Qishenyiqi dropping pills were analyzed on the same C18 reversed-phase column by use of different binary mobile phases. In the chromatogram obtained from sub-chemome I, 19 compounds with strong ultraviolet absorption at 260 nm were detected; 27 compounds with weak ultraviolet absorption at 203 nm were detected in the chromatogram obtained from sub-chemome II. All the compounds were identified by ESI–Q–TOF MS. This study furnished much more important information about the pharmacological activity of Qishenyiqi dropping pills than previous studies and laid a good foundation for further exploration of chemomics.
Co-reporter:Peng Geng, Gang Bai, Xiansheng Meng, Fang Bai, Guoan Luo
Journal of Pharmaceutical and Biomedical Analysis 2009 50(5) pp: 917-923
Publication Date(Web):
DOI:10.1016/j.jpba.2009.06.025
Co-reporter:Peng Geng, Xiansheng Meng, Gang Bai and Guoan Luo
Analytical Chemistry 2008 Volume 80(Issue 19) pp:7554
Publication Date(Web):August 23, 2008
DOI:10.1021/ac801117s
Profiling of acarviostatin family secondary metabolites secreted by Streptomyces coelicoflavus ZG0656 was performed by means of a rapid and facile procedure using ultraperformance liquid chromatography coupled with electrospray ionization mass spectrometry (UPLC/ESI-MS). The acarviostatins were separated on a C18 UPLC column with a series of acetonitrile−aqueous ammonia gradients. The target homologues were detected using the multiple reaction monitoring mode, and the chemical structures were confirmed by analyzing the diagnostic fragment ions in their MS/MS spectra. All six known reference acarviostatins (I03, II03, II13, II23, III03, IV03) were thus identified. In addition, at least 74 acarviostatin homologues, including 65 novel compounds, were characterized. Some of the features of the novel structures included having up to five acarviosine moieties, an acarviosine moiety at the reducing terminus, or an incomplete acarviosine moiety at the nonreducing terminus. This type of investigation may be useful for researchers who study secondary metabolomics in microorganisms and plants, especially those who perform metabolic profiling of aminooligosaccharides and other natural products with similar structures.
Co-reporter:Peng Geng, Feng Qiu, Yuanyuan Zhu, Gang Bai
Carbohydrate Research 2008 Volume 343(Issue 5) pp:882-892
Publication Date(Web):7 April 2008
DOI:10.1016/j.carres.2008.01.020
Four aminooligosaccharides were isolated and purified from the culture filtrate of Streptomyces coelicoflavus ZG0656. Their chemical structures were determined by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and two-dimensional nuclear magnetic resonance (NMR) spectroscopy. The names acarviostatins I03, II03, III03, and IV03 were given to the oligomers due to their acarviosin core structures. Acarviostatins III03 and IV03, which contain three and four acarviosin–glucose moieties, respectively, were identified as novel compounds. The four acarviostatins were all mixed noncompetitive inhibitors of porcine pancreatic α-amylase (PPA). The inhibition constants (Ki) for acarviostatins III03 and IV03 were 0.008 and 0.033 μM, respectively. Acarviostatin III03 is the most effective α-amylase inhibitor known to date, with a Ki value 260 times more potent than acarbose.
Co-reporter:Peng Geng, Gang Bai
Carbohydrate Research 2008 Volume 343(Issue 3) pp:470-476
Publication Date(Web):25 February 2008
DOI:10.1016/j.carres.2007.11.012
Two novel aminooligosaccharides were separated from the culture filtrate of Streptomyces coelicoflavus ZG0656. Their chemical structures were determined by electrospray ionization tandem mass spectrometry (ESI-MS/MS) and 2D nuclear magnetic resonance (NMR) spectroscopy. Because of their acarviosine core structures, the names acarviostatins II23 and II13 were given to the novel compounds. The two acarviostatins were both mixed noncompetitive inhibitors of porcine pancreatic α-amylase (PPA), with inhibition constants (Ki) of 0.009 μM (acarviostatin II23) and 0.010 μM (acarviostatin II13). Therefore, acarviostatin II23 and acarviostatin II13 are, respectively, 231 and 208 times more potent than acarbose.
Co-reporter:Yangsheng Yu;Chunqin Liu;Yang Li;Yongjie Jin
Frontiers in Biology 2007 Volume 2( Issue 4) pp:391-396
Publication Date(Web):2007 October
DOI:10.1007/s11515-007-0059-6
L-cysteine desulfhydrase (CD) plays an important role in L-cysteine decomposition. To identify the CD gene in Pseudomonas sp. TS1138 and investigate its effect on the L-cysteine biosynthetic pathway, the CD gene was cloned from Pseudomonas sp. TS1138 by polymerase chain reaction (PCR) method. The nucleotide sequence of CD gene was determined to be 1,215 bp, and its homology with other sequences encoding CD was analyzed. Then the CD gene was subcloned into pET-21a(+) vector and expressed in Escherichia coli (E. coli) by isopropyl-β-D-thiogalactopyranoside (IPTG) inducement. The recombinant CD was purified by Ni-NTA His-Bind resin, and its activity was identified by the CD activity staining. The enzymatic properties of the recombinant CD were characterized and its critical role involved in the L-cysteine biosynthetic pathway was also discussed.
Co-reporter:Yu Cao, Gang Bai, Jiaqi Chen, Wang Tian, Shenqi Wang, Wenbo Yang
Journal of Chromatography B 2006 Volume 833(Issue 2) pp:236-244
Publication Date(Web):3 April 2006
DOI:10.1016/j.jchromb.2006.02.001
Magnetic agarose microspheres (MAMS), magnetic cellulose microspheres (MCMS), and magnetic poly(vinyl alcohol) microspheres (MPVAMS) were prepared by various different preparation methods. MCMS coupled with anti-IFN α-2b monoclonal antibodies (mAb) were selected for the purification of interferon α-2b (IFN α-2b) after performance characterization among microspheres. Parameters of immunomagnetic separation (IMS), including binding mAb, elution behavior, and sample pretreatment conditions, were optimized to improve the purification efficiency of the separation of IFN α-2b by MCMS. Size-exclusion HPLC (HPSEC) showed that the IFN α-2b was purified from crude cell lysate had an overall purity of 92.9%, while immunological and biological assays showed an activity recovery of 88.5% and specific antiviral activity of 2.7 × 108 IU/mg. Identity and molecular mass of purified IFN α-2b were confirmed by western blot and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) analysis. This study illustrated the favorable separation media which combined desired properties for the development of magnetic separation of biological materials.
Co-reporter:Yuanyuan Hou, Yan Nie, Binfeng Cheng, Jin Tao, Xiaoyao Ma, Min Jiang, Jie Gao, Gang Bai
Acta Pharmaceutica Sinica B (May 2016) Volume 6(Issue 3) pp:212-221
Publication Date(Web):May 2016
DOI:10.1016/j.apsb.2016.03.002
Co-reporter:Xiaohong Qin, Limei Ren, Xue Yang, Fang Bai, Lele Wang, Peng Geng, Gang Bai, Yuequan Shen
Journal of Structural Biology (April 2011) Volume 174(Issue 1) pp:196-202
Publication Date(Web):1 April 2011
DOI:10.1016/j.jsb.2010.11.020
Human pancreatic α-amylase (HPA) catalyzes the hydrolysis of α-d-(1,4) glycosidic linkages in starch and is one of the major therapeutic targets for type II diabetes. Several acarviostatins isolated from Streptomyces coelicoflavus var. nankaiensis previously showed more potent inhibition of HPA than acarbose, which has been successfully used in clinical therapy. However, the molecular mechanisms by which acarviostatins inhibit HPA remains elusive. Here we determined crystal structures of HPA in complexes with a series of acarviostatin inhibitors (I03, II03, III03, and IV03). Structural analyses showed that acarviostatin I03 undergoes a series of hydrolysis and condensation reactions in the HPA active site, similar to acarbose, while acarviostatins II03, III03, and IV03 likely undergo only hydrolysis reactions. On the basis of structural analysis combined with kinetic assays, we demonstrate that the final modified product with seven sugar rings is best suited for occupying the full active site and shows the most efficient inhibition of HPA. Our high resolution structures reported here identify first time an interaction between an inhibitor and subsite-4 of the HPA active site, which we show makes a significant contribution to the inhibitory effect. Our results provide important information for the design of new drugs for the treatment of type II diabetes or obesity.
Co-reporter:Mengge Zhou, Xiaoyao Ma, Guoyu Ding, Zengyong Wang, Dan Liu, Yongling Tong, Hong Zhou, Jie Gao, Yuanyuan Hou, Min Jiang, Gang Bai
Journal of Chromatography B (15 January 2017) Volumes 1041–1042() pp:
Publication Date(Web):15 January 2017
DOI:10.1016/j.jchromb.2016.12.012
•UPLC-Q/TOF MS, PCA and the Hotmap were integrated to clarify the difference of chemical constituents from five BM species.•The antimuscarinic and anti-inflammatory effects of different BM species were firstly compared using our bio-assay.•ANN analysis were used to clarify the active ingredients for each bio-activity.•The emphasis on the bio-activities of various BM species could guild their use in TCM.Many species of Bulbus fritillariae are used as traditional medicines for thousands of years; however, their application is not standardized. To clarify the differences and homologies, the antimuscarinic and anti-inflammatory effects of five BM species were firstly tested and compared at cellular level. With an integrated strategy combining UPLC-Q/TOF MS, PCA and ANN analysis, the active ingredients among 28 different chemical markers were predicted and identified. SB and QB extracts showed the best antimuscarinic effects and several steroidal alkaloids, such as solanidine, contributed to this effects. However, ZB was superior to reduce the inflammatory response. Another five components were responsible by decreasing the expression of NF-κB, including puqiedine, zhepeiresinol, 2-monopalmitin, N-demethylpuqietinone, and isoverticine. More novelty, a new cluster of five BM species based on active ingredients as potential quality markers was depicted to illustrate their functions. These results of the study could make a reference for the medicinal application of BM species in clinic; and the integrated strategy provided an effective method to obtain the quality markers from medical herbs, which was helpful for the quality control of traditional medicinal products.
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![(S)-(1R,3S,5R,6S)-6-Hydroxy-8-methyl-8-azabicyclo[3.2.1]octan-3-yl 3-hydroxy-2-phenylpropanoate](http://img.cochemist.com/ccimg/55900/55869-99-3.png)
![(S)-(1R,3S,5R,6S)-6-Hydroxy-8-methyl-8-azabicyclo[3.2.1]octan-3-yl 3-hydroxy-2-phenylpropanoate](http://img.cochemist.com/ccimg/55900/55869-99-3_b.png)
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![(1R)-7-hydroxy-8-methyl-8-azabicyclo[3.2.1]octane-3,6-diyl (2E,2'E)bis(2-methylbut-2-enoate)](http://img.cochemist.com/ccimg/51300/51261-73-5_b.png)
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![8-azabicyclo[3.2.1]oct-3-yl 3,4-dimethoxybenzoate](http://img.cochemist.com/ccimg/600/537-30-4.png)
![8-azabicyclo[3.2.1]oct-3-yl 3,4-dimethoxybenzoate](http://img.cochemist.com/ccimg/600/537-30-4_b.png)
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