Identification of Chemical Constituents in Chaishi Tuire Granules by UPLC-ESI-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To rapidly identify the chemical constituents of Chaishi Tuire granules by ultra-performance liquid chromatography-electrospray/quadrupole time-of-flight tandem mass spectrometry （UPLC-ESI-Q-TOF-MS/MS）. Method:Chromatographic separation was conducted on a Phenomenex^® Luna omega C₁₈ column （2.1 mm×100 mm， 1.6 μm） with 0.1% formic acid aqueous solution （A）-acetonitrile （B） as the mobile phases for gradient elution （0-20 min， 5%-40%B； 20-40 min， 40%-95%B； 40-43 min， 95%B）， the flow rate was set at 0.3 mL·min^-1. MS data were collected in positive and negative ion modes， the scanning range was <italic>m</italic>/<italic>z</italic> 150-1 500 and electrospray ionization （ESI） was employed. The chemical constituents of Chaishi Tuire granules were identified by comparing with the retention time and the mass data of the reference substances， as well as the accurate mass， MS/MS fragment ions， mass spectrometry databases （PubChem， MassBank， ChemicalBook and others） and related literature. Result:A total of 85 chemical constituents were identified， including 28 flavonoids， 24 phenylpropanoids， 11 terpenoids， 10 alkaloids， 4 quinones， and 8 others. Among them， 19 constituents derived from Lonicerae Japonicae Flos， 14 constituents derived from Scutellariae Radix， 10 constituents derived from Isatidis Radix， 9 constituents derived from Taraxaci Herba， 9 constituents derived from Forsythiae Fructus， 4 constituents derived from Bupleuri Radix， 4 constituents derived from Anemarrhenae Rhizoma， and 4 constituents derived from Rhei Radix et Rhizoma. Conclusion:Chaishi Tuire granules is rich in phytochemicals， which are derived from many of traditional Chinese medicines. This study can lay a foundation for the quality control， material basis and <italic>in vivo</italic> metabolic analysis of this preparation.

The Adaptational Regulation Mechanism of the Apoptosis in Aged Rats’ Striatum in Response to Regular Aerobic Exercise Based on Carbonylation Proteomics / 中国生物化学与分子生物学报

This paper investigated the effects of regular aerobic exercise on protein oxidative stress and apoptosis in aging rat striatum, and further analyzed its target proteins and mechanism based on differential carbonylation proteomics. Totally 24 specific pathogen-free (SPF) 23-month-old male Sprague-Dawley (SD) rats were randomly divided into aged sedentary control group (Con-SED, n = 12) and aged regular aerobic exercise runner group (Aero-EXE, n = 12). The medium intensity of regular aerobic exercise model: The intensity of maximum oxygen consumption (VO

Chemical constituents of Shuangshen Pingfei Granules by UPLC-ESI-Q-TOF-MS/MS / 中草药

Objective: A novel ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was applied to establish a method to recognize and classify the main chemical constituents of Shuangshen Pingfei Granules accurately and rapidly. Methods: ACQUITY UPLC® BEH C18 chromatographic column (100 mm × 2.1 mm, 1.7 μm) was employed to UPLC analysis with the mobile phase consisting of acetonitrile-0.1% formic acid aqueous solution. ESI ion source was used to ensure the data collected in positive and negative ion mode. The chemical components of Shuangshen Pingfei Granules were identified by comparing with the retention time and the mass data of the reference substance, and consulting literature reports and mass spectrometry database. Results: A total of 63 chemical components were identified, including 24 terpenoids, seven phenolic acids, six tanshinones, 14 flavonoids and 12 other classes. Conclusion: The qualitative method established in this study could be used to rapidly and accurately identify the main chemical constituents of Shuangshen Pingfei Granules, and lay a foundation for the further analysis of effective ingredients in vivo, pharmacodynamic material basis and quality control research.

Composition differences of polyphenolic compounds in fruits of Lycium ruthenicum Murr. and Lycium barbarum L / 上海交通大学学报（医学版）

Objective: Both Lycium barbarum L. (LB) and Lycium ruthenicum Murr. (LR) belong to the Lycium genus of Solanaceae family but their fruits have significant phenotypic differences in terms of color and shapes. This study is aimed to investigate the inter-species difference of their fruit polyphenol composition. Methods: The polyphenol composition of fresh and dried fruits of two Lycium species were comprehensively analyzed using ultra-high performance liquid chromatography-electrospray ionization-quadrupole-time of flight mass spectrometry (UHPLC-ESIQ- TOF-MS). Results: 35 polyphenolic compounds were detected and quantified in the fresh and dried Lycium fruits including phenolic acids, anthocyanins and phenolamides (i.e., the amide-adducts of polyphenolic acids and polyamines). For both species, the dried fruits had more polyphenolic compounds than their fresh fruits with significant inter-species difference; for both fresh and dried fruits, LR had more polyphenolic compounds than LB. Conclusion: There are significant inter-species differences between these two Lycium in both fresh and dried fruits. These results offer essential information for further understanding of the biological functions of these two Lycium fruits as phytomedicines and functional food.

Chemical constituent analysis for seeds of Celosia argentea by UPLC-ESI-Q-TOF-MS / 中国中药杂志

This Paper aimed to analyze and identify the chemical constituents from the seeds of Celosia argentea by UPLC-ESI-Q-TOF-MS. The analysis was performed on an ACQUITY HSS T3 reverse phase column(2.1 mm ×100 mm, 1.8 μm). The mobile phase consisting of 0.1% formic acid acetonitrile and 0.1% aqueous formic acid was used for gradient elution, and the flow rate was 0.4 mL·min~(-1). Mass spectrometry was applied for the qualitative analysis under positive and negative ionization modes and ESI ion source. Data was analyzed by Masslynx 4.1 software, literatures in SciFinder database, and standards. A total of 49 compounds, including 14 triterpenoids, 17 flavonoids, 11 cyclic peptides, 2 phenols, 2 organic acids, and 3 steroids were putatively identified. Among them, 19 compounds were firstly reported from this species. In-depth chemical constituent analysis for the seeds of C. argentea were accomplished here, and the findings could lay a good foundation for its quality control and clarifying the material basis of its efficacy.

Rapid identification of constituents of Urtica hyperborea using UPLC-ESI-Q-TOF-MS/MS method / 中国中药杂志

This paper deals with the application of ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry(UPLC-ESI-Q-TOF-MS/MS) method to rapidly determine and analyze the chemical constituents of methanol extract of Urtica hyperborea. We employed UPLC YMC-Triart C18(2. 1 mm×100 mm,1. 9 μm) column to UPLC analysis with acetonitrile-water(containing 0. 4% formic acid) in gradient as mobile phase. The flow rate was 0. 3 m L·min-1 gradient elution and column temperature was 30℃; the injection volume was 4 μL. ESI ion source was used to ensure the data collected in anegative ion mode. The chemical components of U. hyperborea were identified through retention time,exact relative molecular mass,cleavage fragments of MS/MS and reported data.The results indicated that a total of 31 compounds were identified,including 8 flavonoids,14 phenolic compounds,8 phenylpropanoids(4 coumarins and 4 lignans),and 1 steroidal compound,13 of which were confirmed by comparison. The UPLC-ESI-Q-TOF-MS/MS method could rapid identify the chemical components of U. hyperborea. The above compounds were discovered in U. hyperborea for the first time,which could provide theoretical foundation for further research on the basis of the pharmacodynamics of U. hyperborea.

Identification and characterization of major constituents in Juglans mandshurica using ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-ESI-Q-TOF/MS) / 中国天然药物

The present study was designed to identify and characterize the major constituents in Juglans mandshurica Maxim. A simple, efficient and sensitive ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-ESI-Q-TOF/MS) method was established and validated under positive and negative ion modes. The separation was performed on a Waters ACQUITY UPLC BEH C column (2.1 mm × 100 mm, 1.7 μm) by gradient elution with a mobile phase (Phase A: 0.1% aqueous formic acid solution, Phase B: 0.1% formic acid acetonitrile solution). A total of 165 compounds were rapidly selected by Targeted and Non-Targeted Peak Finding approaches, and then tentatively identifled by comparing with reference substances or inferred through mass spectrometry fragment ion analysis and literature data. These compounds included 68 naphthalenequinones, 20 diarylheptanoids, 29 flavonoids, 20 triterpenes, and 28 phenolic acids. In conclusion, the present study provided an effective approach to identifying components in complex matrices of herbal medicines such as Juglans mandshurica Maxim.

Identification and characterization of major constituents in Juglans mandshurica using ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-ESI-Q-TOF/MS) / 中国天然药物

The present study was designed to identify and characterize the major constituents in Juglans mandshurica Maxim. A simple, efficient and sensitive ultra performance liquid chromatography coupled with time-of-flight mass spectrometry (UPLC-ESI-Q-TOF/MS) method was established and validated under positive and negative ion modes. The separation was performed on a Waters ACQUITY UPLC BEH C column (2.1 mm × 100 mm, 1.7 μm) by gradient elution with a mobile phase (Phase A: 0.1% aqueous formic acid solution, Phase B: 0.1% formic acid acetonitrile solution). A total of 165 compounds were rapidly selected by Targeted and Non-Targeted Peak Finding approaches, and then tentatively identifled by comparing with reference substances or inferred through mass spectrometry fragment ion analysis and literature data. These compounds included 68 naphthalenequinones, 20 diarylheptanoids, 29 flavonoids, 20 triterpenes, and 28 phenolic acids. In conclusion, the present study provided an effective approach to identifying components in complex matrices of herbal medicines such as Juglans mandshurica Maxim.

Analysis on chemical constituents from Swertia mileensis by UPLC-ESI-Q-TOF-MS / 中草药

Objective: To analyze and identify the chemical constituents from Swertia mileensis by UPLC-ESI-Q-TOF-MS. Methods: The analysis was performed on an Acquity HSS T3 reverse phase column (100 mm × 2.1 mm, 1.8 μm). The mobile phase consisted of 0.1% formic acid acetonitrile and 0.1% formic acid, and was used for gradient elution, with the flow rate of 0.4 mL/min. Mass spectrometry was applied for the qualitative analysis under positive and negative ion modes and ESI ion source. Data was analyzed by Masslynx 4.1 software, SciFinder database, literatures, and standards. Results: Twenty-eight compounds, including 7 iridoids, 14 xanthones, 3 flavonoids, 2 triterpenes, and 2 phenols were identified from S. mileensis. Among them, vogeloside, 8-O-β-D- glueopyranosyl-(l→6)-β-D-glueopyranosyl-l,7-dihydroxy-3-ethoxyxanthone, 3-O-demethylswertipunicoside, and sweriyunnanlactone A were reported from this species for the first time. Conclusion: Using UPLC-ESI-Q-TOF-MS method the main chemical constituents from S. mileensis can be rapidly and accurately identified, which provides a new strategy for its quality control as well as a reference for clarifying the material basis of its efficacy.

Determination of chemical constituents in Lonicera maackii with HPLC-DAD-ESI-Q-TOF/MS / 中草药

Objective: To identify and determine the main chemical constituents of Lonicera maackii, and to provide data support for the development and utilization of L. maackii. Methods: The chemical constituents of L. maackii were identified by HPLC-DAD-ESI-Q-TOF/MS and the main chemical constituents were determined by HPLC-DAD. Results: A total of 31 chromatographic peaks were detected and 19 chemical constituents were identified from L. maackii, and the contents of the 10 compounds (logaric acid, chlorogenic acid, loganin, morroniside, Secoxyloganin, rutin, hyperoside, luteoloside, chlorogenic acid A, and chlorogenic acid C) were determined. Conclusion: The contents of the main chemical constituents in the flower buds of L. maackii were higher than those in the blooming stage and yellow flowering stage. The contents of main components in L. maackii were similar to those of Lonicerae Japonicae Flos. The results of this study provide data support for the development and utilization of L. maackii.

Metabolism of effective fractions of Polygonum capitatum in rat feces and bile / 中草药

Objective: To investigate the metabolite profile of effective fractions of Polygonum capitatum in rat feces and bile. Methods: In this study, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC- ESI-Q/TOF-MS) was applied to the identification of metabolites of P. capitatum extracts in rat bile and feces after ig administration. Mass defect filter and Metabolite Tools software were also used. Results: The four parent components and 26 metabolites were identified in rats metabolic experiments in vivo, including 12 metabolites in feces and 19 metabolites in bile. The major metabolites were glucuronide conjugation, methyl-glucuronide conjugation, and sulfate conjugation of quercetin; Conclusion: The studies have established a high-resolution mass spectrometry method to analyze the metabolites of effective fractions of P. capitatum in vivo. The in vivo metabolic characteristics of the components presented in the fraction have been clarified. The work lays a foundation for the clinical application and development of P. capitatum.

Analysis of the Chemical Compositions in Jinqiancao Granule by HPLC-ESI-Q-TOF-MS / 中国药房

OBJECTIVE:To establish a method for the analysis of chemical compositions in Jinqiancao granule. METHODS:HPLC-ESI-Q-TOF-MS was adopted. The chromatographic conditions:the column was Inertsil ODS-4 with mobile phase of 0.1%formic acid- methanol (gradient elution) at a flow rate of 0.8 ml/min,column temperature was 30 ℃,and the injection volume was 15 μl. MS conditions:ion source was ESI(negative ion mode),endplate offset voltage was -500 V,capillary electrophore-sis was 3500 V,carrier gas was helium gas,atomization and drying gas was high purity nitrogen gas at a flow rate of 6 L/min and a pressure of 1.0 bar,drying air temperature was 180℃. Scanning range was 50-1500 m/z. ChemBioDraw Ultra13.0 and Bruk-er data analysis software 4.0 were used to analyze the chemical composition molecular formula. RESULTS:A total of 27 kinds of chemical components were identified in Jinqiancao granule,involving sucrose,uridine,gallic acid,new chlorogenic acid,(-)-epi-gallocatechin,protocatechuic acid,chlorogenic acid,schaftoside,caffeic acid,schaftoside,vanillc acid,ferulic acid,hyperin,ros-marinic acid,rutin,isoquercitrin,astragalin,quercetin-3-rhamnoside,myricetin,4-hydroxybenzoic acid,quercetin,naringenin, luteolin,kaempferol,isorhmnetin,apigenin and emodin. CONCLUSIONS:Caffeic acid,rosmarinic acid,vanillic acid,(-)-epigal-locatechin,uridine and emodin are firstly found and reported in the chemical compositions in Jinqiancao granule.

Qualitative Analysis and Quality Evaluation of Cnidium monnieri Using UHPLC-ESI-Q-TOF/MS / 中草药·英文版

Objective To identify the coumarins constituents in Cnidium monnieri and classify ten samples into three groups and this helpful chemical information could be used for the further pharmacological and clinical study on C. monnieri. Methods Qualitative analysis of coumarins in C. monnieri was detected by UHPLC-ESI-Q-TOF/MS. Quadrupole TOF/MS in either full scan mode or extracted ion mode was used for the qualitative analysis of the constituents. Relative peak area of each component was used for the hierarchical cluster analysis. Results According to UHPLC-ESI-QTOF-MS data, chemical structures of 28 coumarins in the fruits of C. monnieri were identified, including 19 simple coumarins, seven linear coumarins, and two angular coumarins. Among these constituents, ten coumarins were firstly identified in C. monnieri. In addition, hierarchical cluster analysis suggested that C. monnieri from different regions could be classified into four groups, and this clustering was correlated to the distribution significantly, Xuzhou could be regarded as the genuine producing area. Conclusion UHPLC-ESI-Q-TOF-MS is a viable method for qualitative analysis and quality evaluation of coumarins from the fruit of C. monnieri. Coumarins in C. monnieri exists intra-species variance, which indicates significant meaning for the quality control and choice of famous region drug for C. monnieri in the clinic medication.

Chemical constituents from water extract of Toosendan Fructus by UPLC-ESI-Q-TOF-MS / 中草药

Objective: To analyze and identify the chemical constituents from the water extract of Toosendan Fructus by UPLC-ESI- Q-TOF-MS. The analysis was performed on an ACQUITY HSS T3 reverse phase column (2.1 mm × 100 mm, 1.8 μm). The mobile phase consisted of acetonitrile and 0.1% formic acid was used for gradient elution, the flow rate was 0.4 mL/min. Methods: Quadrupole-time of flight-mass spectrometry was applied for the qualitative analysis under positive and negative ion modes and ESI ion source was used for mass spectra. Results: The results indicated that fifteen compounds from the water extract of Toosendan Fructus had been identified by direct comparison in both positive and negative ion mass data, the element compositions analysis, and the data of the literature. They are vanillic acid, lilac acid, p-hydroxybenzoic acid, rutin, meliatoosenin E, toosendanin, Δ5,6-isotoosendanin, isotoosendanin, meliatoosenin N, meliatoosenin P, 1-deacetylnimbolinin B, meliatoosenin R, 1-O-tigloyl-1-O-debenzoylohchinal, meliatoosenin R, and nimbolinin B. Conclusion: The efficient separation ability of UPLC and high sensitive detection of MS are used in this study, which will provide the evidences for evaluating the quality of Toosendan Fructus herbs, stabilizing the curative effect in clinic, and explaining the mechanism.

Diagnostic ion filtering strategy for chemical characterization of Guge Fengtong Tablet with high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry / 中国天然药物

The present study was designed to characterize the chemical constituents of Guge Fengtong Tablet (GGFTT). Based on the chromatographic retention behavior, fragmentation pathways of chemical components and the published literatures, a diagnostic ion filtering strategy with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-Q-TOF/MS) was established to identify the multiple bioactive constituents of GGFTT. The rapid identification of forty-seven components, including 18 phenolic acids, 8 saponins, 14 gingerol-related compounds, and 7 diarylhepatonoids, was accomplished using this newly developed method. The coupling of HPLC-ESI-Q-TOF/MS with the diagnostic ion filtering strategy was useful and efficient for the in-depth structural elucidation of chemical compounds of GGFTT.

Determination of Protopine in Rat Brain Tissues by RRLC-ESI/Q-TOF-MS Method / 中草药·英文版

Objective: To analyse the quantification of protopine from Corydalis Decumbentis Rhizoma (CDR) extract in brain tissues of rats. Methods: A rapid, sensitive, and accurate analytical method based on rapid resolution liquid chromatography (RRLC) coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (Q-TOF-MS) was developed for the quantification of protopine in brain of rats. A simple liquid-liquid extraction process was employed for the sample preparation. Chromatographic separation was achieved using 1.8 μm porous particle columns. Results: The calibration curve of protopine was linear in the range of 12-897 ng/mL. The relative standard deviations of intra- and inter-day precision values were less than 10%. The extraction recoveries were 96.4%, 99.6%, and 98.5%, for protopine at the concentration of 598.0, 119.6, and 12.0 ng/mL, respectively, and internal standard (1.27 μg/mL) was (98.60 ± 0.02)%. Conclusion: The validated method is successfully applied for the determination of protopine in brain tissue of rats after ig administration of CDR extract. © 2014 Tianjin Press of Chinese Herbal Medicines.

Selective separation, detection of zotepine and mass spectral characterization of degradants by LC-MS/MS/QTOF / 药物分析学报

A simple, precise, accurate stability-indicating gradient reversed-phase high-performance liquid chromatographic (RP-HPLC) method was developed for the quantitative determination of zotepine (ZTP) in bulk and pharmaceutical dosage forms in the presence of its degradation products (DPs). The method was developed using Phenomenex C18 column (250 mm ~ 4.6 mm i.d., 5 mm) with a mobile phase containing a gradient mixture of solvents, A (0.05%trifluoroacetic acid (TFA), pH ? 3.0) and B (acetonitrile). The eluted compounds were monitored at 254 nm;the run time was within 20.0 min, in which ZTP and its DPs were well separated, with a resolution of 41.5. The stress testing of ZTP was carried out under acidic, alkaline, neutral hydrolysis, oxidative, photolytic and thermal stress conditions. ZTP was found to degrade significantly in acidic, photolytic, thermal and oxidative stress conditions and remain stable in basic and neutral conditions. The developed method was validated with respect to specificity, linearity, limit of detection, limit of quantification, accuracy, precision and robustness as per ICH guidelines. This method was also suitable for the assay determination of ZTP in pharmaceutical dosage forms. The DPs were characterized by LC-MS/MS and their fragmentation pathways were proposed.

Structural characterization and identification of major constituents in radix scrophulariae by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry / 中国天然药物

AIM@#To analyze the major constituents in Radix Scrophulariae (Scrophularia ningpoensis).@*METHOD@#Radix Scrophulariae was analyzed by HPLC coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (ESI-Q-TOF MS/MS). Compounds were separated by HPLC using a C18 column and gradient elution of acetonitrile and 0.1 % (V/V) acetic acid-water. Negative ion mode was employed.@*RESULTS@#A total of thirty-six compounds, including fourteen iridoid glycosides, nineteen phenylpropanoid glycosides, and three organic acids, were identified from Radix Scrophulariae based on the accurate mass measurement of precursor and product ions. Twenty-one of the constituents were identified by comparing their retention times (tR) and ESI-MS/MS data with those of reference standards and/or previous publications, while another fifteen compounds were tentatively identified or deduced according to their Q-TOF MS/MS data which afforded sufficient structural information.@*CONCLUSION@#It is believed that this study is useful for the identification of constituents in Radix Scrophulariae, as well as related plants and complex prescriptions.

Chemical profiling of Jinqi Jiangtang tablets by HPLC-ESI-Q-TOF/MS / 中国天然药物

AIM@#To profile the chemical constituents in Jinqi Jiangtang tablets.@*METHOD@#Based on the chromatographic retention behavior, fragmentation patterns of chemical components, and published literatures, a high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-Q-TOF/MS) method was established to characterize and identify components in Jinqi Jiangtang tablets.@*RESULTS@#A total of 52 chemical compounds, including eight iridoid glycosides, seven phenolic acids, twelve alkaloids, six flavonoids, and nineteen saponins, were identified in Jinqi Jiangtang tablets.@*CONCLUSION@#The established method could serve as a powerful tool for structural characterization and quality control of this Chinese herbal preparation.

Comparative Proteomics Analysis of LCM Purified Nasopharyngeal Carcinoma and Normal Nasopharyngeal Epithelial Tissue / 生物化学与生物物理进展

To search for nasopharyngeal carcinoma (NPC) biomarkers,laser capture microdissection (LCM) was used to purify the target cells from NPC and normal nasopharyngeal epithelial tissues (NNET). Two-dimensional gel electrophoresis (2-DE) was performed to separate the total proteins of microdissected NPC and NNEC,PDQuest software was applied to analyze 2-DE images,and the differential protein spots between the two types of tissues were identified by both MALDI-TOF-MS and ESI-Q-TOF-MS. The expression of cytokeratin 8(CK8),one of the differential proteins,in the microdissected NPC and NNEC as well as 4 NPC cell lines with different differentiated degrees and/or metastatic potentials was detected by Western blot. Immunohistochemistry was also used to detect the expression of CK8 in paraffin-embedded tissues including 63 cases of primary NPC,28 cases of NNET and 20 cases of cervical lymphonode metastasis. In the present study,2-DE patterns of microdissected NPC and NNEC were established,and 29 differential proteins in the above two tissues were identified,of which 15 only expressed or up-regulated in NPC and 14 only expressed or up-regulated in NNET. The expression level of differential protein CK8 between the NPC and NNET was selectively confirmed,and was found to be related to the differentiation and/or metastasis of NPC cell lines. Significant down-regulation of CK8 was observed in NPC compared with NNET,and significant up-regulation of CK8 was also observed in lymphonode metastasis compared with primary NPC. The data suggest that CK8 may be related to the differentiation and lymphonode metastasis of NPC,and may serve as molecular biomarkers for metastasis and differentiation of NPC.