Analysis on Components Absorbed into Blood and Cerebrospinal Fluid of Schisandrae Chinensis Fructus Based on Sequential Metabolism / 中国实验方剂学杂志

ObjectiveTo identify the prototypical components and metabolites absorbed into blood and cerebrospinal fluid of Schisandrae Chinensis Fructus（SCF） based on sequential metabolism combined with liquid chromatography-mass spectrometry. MethodBlood and cerebrospinal fluid samples of integrated metabolism， intestinal metabolism and hepatic metabolism were collected from male SD rats after gavage and in situ intestinal perfusion administration， and ultra-performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry（UPLC Q-Exactive Orbitrap MS） was used to analyze and compare the differences in the spectra of SCF extract， blank plasma， administered plasma， blank cerebrospinal fluid and administered cerebrospinal fluid with ACQUITY UPLC BEH Shield RP18 column（2.1 mm×100 mm， 1.7 µm）， the mobile phase was acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-7 min， 95%B； 7-12 min， 95%-35%B； 12-17 min， 35%-15%B； 17-20 min， 15%-12%B； 20-22 min， 12%-5%B； 22-23 min， 5%B； 23-25 min， 5%-95%B； 25-28 min， 95%B）. And heated electrospray ionization（HESI） was used with positive and negative ion modes， the scanning range was m/z 100-1 500. The prototypical constituents and their metabolites absorbed into blood and cerebrospinal fluid of SCF were identified according to the retention time， characteristic fragments， molecular formulae and the information of reference substances. ResultA total of 42 chemical components were identified in the extract of SCF， including lignans， flavonoids， amino acids， tannins， and others， of which lignans were the main ones. A total of 27 prototypical components and 14 metabolites were identified in plasma samples from different sites. A total of 15 prototypical components and 9 metabolites were identified in cerebrospinal fluid. The main metabolic reactions involved in the formation of metabolites were mainly demethylation， methylation， demethoxylation and hydroxylation. ConclusionThrough the systematic identification of the prototypical components and metabolites of SCF in rats， it provides data support for further better exploring the material basis of SCF in the treatment of central nervous system diseases.

Characterization of Effective Components in Guizhi Jia Gegentang for Intervention of Influenza Virus Pneumonia Based on Serum Pharmacochemistry of Traditional Chinese Medicine / 中国实验方剂学杂志

ObjectiveTo characterize the efficacy components of Guizhi Jia Gegentang（GGT） in intervening influenza virus pneumonia by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap MS）. MethodBALB/c mice were randomly divided into normal group and GGT group（36 g·kg-1·d-1） with six mice in each group. GGT group was continuously administered GGT extract for 5 d， while the normal group was administered an equal amount of ultrapure water. Serum and lung tissue were collected after administration， and UPLC-Q-Exactive Orbitrap MS was used to characterize the prototypical and metabolic components of GGT in serum and lung tissue of mice. The components existed simultaneously in the serum and lung tissue of mice from the GGT group were defined as its functional components， and the targets of efficacy components were searched by SwissTargetPrediction database， and GeneCards database was used to query the target of influenza virus pneumonia， and then the intersection was taken to obtain potential targets of GGT for intervening in the disease. Protein-protein interaction（PPI） network analysis of potential targets was performed by STRING database， and Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway enrichment analysis on potential targets was performed by Metascape. ResultA total of 29 prototypical components and 28 metabolic components of GGT were detected in the drug-containing serum of mice， of which 11 prototypical components and 4 metabolic components were detected in the lung tissue of mice. The main metabolic pathways included reduction， hydroxylation， methylation， glucuronidation and sulfation. The results of PPI network and KEGG analysis showed that these functional components may act through their effects on targets such as albumin（ALB）， epidermal growth factor receptor（EGFR）， steroid receptor coactivator（SRC）， Toll-like receptor 4（TLR4）， nuclear transcription factor（NF）-κB and adhesion junction. ConclusionThe 11 prototypical components and 4 metabolites present simultaneously in the drug-containing serum and lung tissue of mice may be the potential therapeutic components of GGT in interfering with influenza viral pneumonia， and act through interfering with inflammatory metabolic pathways. This study can provide a reference for the mechanism study of GGT in the treatment of influenza viral pneumonia.

Determination of antibiotic residues in chicken meat using ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry / 预防医学

Objective@#To optimize the sample pretreatment and establish an ultra-high-performance liquid chromatography coupled with hybrid quadrupole-orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap-MS) assay based on the parallel reaction monitoring (PRM) mode for determination of antibiotic residues in chicken meat.@*Methods@#Blank matrix-spiked chicken meat samples were extracted with 95% acetonitrile aqueous solution containing Na2EDTA and formic acid. The extraction solutions were cleaned up using different combinations of C18, PSA and GCB fillers, and the combinations with a higher antibiotic recovery rate was screened. The residues of 32 antibiotics were determined using UPLC-Q-Orbitrap-MS based on the PRM mode.@*Results@#If the extraction solution was cleaned up using the C18 filler, the largest number of antibiotics with a spiked recovery rate of >80% was seen, with matrix effects of 82.2% to 112.6%. The detection limits of 32 antibiotics were 0.8 to 5.8 μg/kg, with linear correlation coefficients of >0.99, spiked recovery rates of 71.3% to 111.5% and relative standard deviations of 3.2% to 14.2%.@*Conclusion@#The UPLC-Q-Orbitrap-MS assay is suitable for determination and quantitative analysis of multiple antibiotics in chicken meat. Key words： high-resolution mass spectrometry orbitrap antibiotic residue parallel reaction monitoring

Analysis on Metabolites and Metabolic Pathways of Limonin in Rats by UHPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

ObjectiveUltra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS） was used to identify the metabolites of limonin in rats， and to explore the gender differences in the distribution of prototype components and metabolites in rats after single dose intragastric administration of limonin， as well as to speculate the metabolic pathways. MethodThe separation was performed on a Thermo Scientific Accucore™ C18 column（3 mm×100 mm， 2.6 μm） with 0.1% formic acid aqueous solution（A）-0.1% formic acid acetonitrile solution（B） as mobile phase in a gradient elution mode（0-1 min， 5%B； 1-6 min， 5%-20%B； 6-18 min， 20%-50%B； 18-23 min， 50%-80%B； 23-25 min， 80%-95%B； 25-30 min， 95%B） at a flow rate of 0.3 mL·min-1 and a column temperature of 30 ℃. MS data of biological samples were collected under the positive ion mode of electrospray ionization（ESI） and in the scanning range of m/z 100-1 500. Plasma， tissues（heart， liver， spleen， lung， kidney， stomach and small intestine）， urine and fecal samples were collected and prepared after intragastric administration， and the prototype component and metabolites of limonin were identified. ResultThe prototype component of limonin were detected in the feces， stomach， small intestine of female and male rats， and in the heart， liver， spleen， lung and kidney tissues of female rats. A total of 23 metabolites related to limonin were detected in rats， of which 2， 1， 5， 4， 23 metabolites were detected in liver， stomach， small intestine， urine and feces， respectively， and the main metabolic pathways were hydrolysis， reduction， hydroxylation and methylation， etc. The distribution of some metabolites differed between male and female rats. ConclusionThe prototype component of limonin are mainly distributed in the stomach and small intestine in rats， and the distribution of prototype component and some metabolites are different by gender. Limonin is mainly excreted through feces with phase Ⅰ metabolites as the main ones. The results of this study can provide a reference for further elucidation of the effect of gender differences on the metabolism of limonin in vivo and its mechanism of action.

Chemical Constituent Analysis of Tongxie Yaofang Decoction by UPLC-LTQ-Orbitrap-MS / 中国实验方剂学杂志

ObjectiveTo rapidly identify the chemical constituents in Tongxie Yaofang decoction by ultra-performance liquid chromatography-linear ion trap-electrostatic field orbitrap high-resolution mass spectrometry（UPLC-LTQ-Orbitrap-MS）. MethodChromatographic conditions were ACQUITY UPLC BEH C18 column（2.1 mm×100 mm， 1.7 μm）， mobile phase of 0.1% formic acid aqueous solution（A）-acetonitrile（B） for gradient elution （0-4 min， 5%-15%B； 4-10 min， 15%-25%B； 10-15 min， 25%-60%B； 15-20 min， 60%-90%B； 20-25 min， 90%-100%B； 25-27 min， 100%B； 27-30 min， 100%-5%B； 30-32 min， 5%B）， flow rate of 0.3 mL·min-1， column temperature at 35 ℃ and injection volume of 3 μL. UPLC-LTQ-Orbitrap-MS was equipped with an electrospray ionization（ESI）， the MS and MS/MS data were collected in positive and negative ion modes， and detection range was m/z 100-1 250. Combining the reference substance， chemical databases and related literature information， TraceFinder 4.1 and Xcalibur 2.1 were used to identify the chemical constituents of Tongxie Yaofang decoction. ResultA total of 90 compounds， mainly including flavonoids， coumarins， monoterpene glycosides， chromones and lactones， were identified from Tongxie Yaofang decoction. By attributing the sources of Chinese medicines for all identified compounds， 9 of them were found to be derived from Atractylodis Macrocephalae Rhizoma， 21 from Paeoniae Radix Alba， 24 from Citri Reticulatae Pericarpium， 29 from Saposhnikoviae Radix， and 7 from at least two Chinese medicines. ConclusionThe method can effectively， quickly and comprehensively identify the chemical components of Tongxie Yaofang decoction， and clarify the chemical composition. These identified compounds cover the main active ingredients of the four herbs with high abundance， which indicates that the extraction method and the ratio of the medicinal materials of Tongxie Yaofang are scientific， and can provide a reference for the research on the material basis and quality evaluation of this famous classical formula.

Analysis on Components Absorbed into Blood of Citri Reticulatae Pericarpium in Rats Based on Sequential Metabolism and UPLC-HRMS / 中国实验方剂学杂志

ObjectiveTo study the metabolism of chemical components from Citri Reticulatae Pericarpium（CRP）in different parts of rats by sequential metabolism and ultra performance liquid chromatography-high resolution mass spectrometry（UPLC-HRMS）. MethodSD male rats were employed as experimental subjects， and blood samples of intestinal metabolism and hepatic metabolism were prepared after administration of CRP ethanol extract by in situ intestinal perfusion， and comprehensive metabolic samples were collected after intragastric administration. UPLC-HRMS was used to analyze the samples with acetonitrile（A）-0.1% formic acid aqueous solution（B）as the mobile phase for gradient elution（0-10 min， 10%-30%A； 10-30 min， 30%-95%A； 30-31 min， 95%-10%A； 31-35 min， 10%A）at a flow rate of 0.35 mL·min-1， using a heated electrospray ionization with positive and negative ion mode scanning in the range of m/z 100-1 500. Under these conditions， the differences in the profiles of CRP ethanol extract， blank plasma and drug-containing plasma under different treatment groups were compared， and the chemical components of each sample were analyzed and identified based on the retention time， accurate relative molecular mass， primary and secondary ion fragments， and the information of reference substances. ResultA total of 44 chemical components were identified in the CRP ethanol extract， including flavone-O-glycosides， flavone-C-glycosides and polymethoxyflavonoids， etc. The results of sequential metabolism showed that 22 chemical components in CRP were detected in the intestinal metabolic sample， 18 chemical components were detected in the hepatic metabolic sample， and 9 identical chemical components（narirutin， hesperidin， meranzin， 5，7，8，3ʹ，4ʹ，5ʹ-hexamethoxy-flavone， isosinensetin， sinensetin， 3，5，6，7，8，3ʹ，4ʹ-heptamethoxyflavone， nobiletin and tangeretin）could be detected in all three metabolic samples， with a total of 22 compounds entering the blood in prototype form. ConclusionThe identified 21 components with well-defined structures entering the blood as prototypes may be potential active components of CRP， and differences in the components at different metabolic parts can provide an experimental basis for elucidating the in vivo biotransformation process of the metabolic components of CRP.

Structural Analysis and Characterization of 4-F-α-PVP Analog 4-F-3-Methyl-α- PVP Hydrochloride / 法医学杂志

OBJECTIVES@#To identify 1-(4-fluorophenyl)-2-(1-pyrrolidinyl) pentan-1-one (4-F-α-PVP) analog 1-(4-fluoro-3-methyl phenyl)-2-(1-pyrrolidinyl) pentan-1-one (4-F-3-Methyl-α-PVP) hydrochloride without reference substance.@*METHODS@#The direct-injection electron ionization-mass spectrometry (EI-MS), GC-MS, electrospray ionization-high resolution mass spectrometry (ESI-HRMS), ultra-high performance liquid chromatography-high resolution tandem mass spectrometry (UPLC-HRMS/MS), nuclear magnetic resonance (NMR), ion chromatography and Fourier transform infrared spectroscopy (FTIR) were integrated utilized to achieve the structural analysis and characterization of the unknown compound in the sample, and the cleavage mechanism of the fragment ions was deduced by EI-MS and UPLC-HRMS/MS.@*RESULTS@#By analyzing the direct-injection EI-MS, GC-MS, ESI-HRMS and UPLC-HRMS/MS of the compound in the samples, it was concluded that the unknown compound was a structural analog of 4-F-α-PVP, possibly with one more methyl group in the benzene ring. According to the analysis results of 1H-NMR and 13C-NMR, it was further proved that the methyl group is located at the 3-position of the benzene ring. Since the actual number of hydrogen in 1H-NMR analysis was one more than 4-F-3-Methyl-α-PVP neutral molecule, it was inferred that the compound existed in the form of salt. Ion chromatography analysis results showed that the compound contained chlorine anion (content 11.14%-11.16%), with the structural analysis of main functional group information by FTIR, the unknown compound was finally determined to be 4-F-3-Methyl-α-PVP hydrochloride.@*CONCLUSIONS@#A comprehensive method using EI-MS, GC-MS, ESI-HRMS, UPLC-HRMS/MS, NMR, ion chromatography and FTIR to identify 4-F-3-Methyl-α-PVP hydrochloride in samples is established, which will be helpful for the forensic science laboratory to identify this compound or other analog compounds.

Comparison of Effect of Ancient and Modern Processing Methods on Chemical Components of Lilii Bulbus Decoction Based on UHPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

ObjectiveTo compare the effects of different processing methods in ancient and modern times on the chemical components of Lilii Bulbus decoction， and to provide experimental support for the origin processing， decoction piece processing and clinical application of this herb. MethodUltra high performance liquid chromatography tandem quadrupole electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS） was used for structural identification of the compounds using excimer ions， secondary MS and characteristic fragment ions， and referring to relevant literature and database information. Principal component analysis（PCA） and orthogonal partial least squares discriminant analysis（OPLS-DA） were used to screen the main differential components， the differential components were quantitatively studied by high performance liquid chromatography（HPLC）， in order to compare the types and contents of chemical components in the decoction of different processing products of Lilii Bulbus. ResultA total of 24 chemical components were identified from the decoction of different processed products of Lilii Bulbus， water extract and scalding liquid of fresh Lilii Bulbus， including 17 phenols， 5 saponins and 2 alkaloids. Compared with the fresh Lilii Bulbus decoction， the contents of regaloside A， p-coumaric acid， colchicine and other components in the decoction of dry Lilii Bulbus processed by scalding method decreased， the content of regaloside C in the decoction of dry Lilii Bulbus processed by steaming method decreased， and the contents of regaloside A and regaloside C in the decoction of fresh Lilii Bulbus processed by water immersion also decreased. Compared with the decoction of dry Lilii Bulbus processed by scalding method， the overall content of components in the fresh Lilii Bulbus decoction and the decoction of fresh Lilii Bulbus processed by water immersion was higher， the contents of components in the decoction of dry Lilii Bulbus processed by steaming method was higher， except for the slightly lower content of regaloside C. ConclusionDifferent processing processes have a certain effect on the types and contents of chemical components in Lilii Bulbus decoction. Scalding process is beneficial to the preservation of Lilii Bulbus， but can cause the loss of effective components. Compared with scalding method， steaming method can prevent browning of Lilii Bulbus and reduce the loss of its active ingredients. The processing method of removing foam after overnight immersion proposed by ZHANG Zhongjing may be more conducive to the treatment of Baihe disease， which can provide reference for the clinical rational application and mechanism research of different processed products of Lilii Bulbus.

Non-targeted screening of emerging contaminants in drinking water based on high-resolution mass spectrometry / 上海预防医学

ObjectiveTo establish a non-targeted screening method for emerging contaminants in drinking water based on high-resolution mass spectrometry and apply it to actual water samples. MethodsA total of 9 drinking water samples collected from 3 reservoirs in Shanghai were purified and concentrated by HLB solid phase extraction column， then separated and analyzed by liquid chromatography high-resolution mass spectrometer and gas chromatography high⁃resolution mass spectrometer. The acquired data were analyzed by Thermo Tracefinder， Excel and other software combined with mzCloud and NIST databases. The methodology was verified with representative compound standards. Pesticide and perfluorinated compounds were taken as examples to analyze their pollution status. ResultsA non-targeted analysis strategy based on liquid chromatography and gas chromatography tandem high-resolution mass spectrometry was established. The pollution level of 20 kinds of pesticides and 4 kinds of perfluorinated compounds identified in 9 drinking water samples were higher in the Huangpu River than in the Yangtze River estuary. ConclusionThe established non-targeted screening method by high-resolution mass spectrometry can detect potential emerging contaminants in drinking water without relying on the standards， which provides a powerful technical means for water quality monitoring and risk assessment.

Exploring of Transformation Rule of Saponins in Polygalae Radix Before and After Processing Based on Simulated Processing Technology / 中国实验方剂学杂志

ObjectiveTo investigate the transformation mechanism and content variation of saponins from Polygalae Radix before and after being boiled with licorice juice and water. MethodSimulated licorice juice boiled products and simulated water boiled products of onjisaponin B， onjisaponin Z， onjisaponin F， polygalasaponin ⅩⅩⅧ were prepared by simulated processing technology， and analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UPLC-Q-Exactive Orbitrap/MS）. Then the contents of onjisaponin B， onjisaponin Z， onjisaponin F， polygalasaponin ⅩⅩⅧ and tenuifolin in Polygalae Radix， licorice-boiled Polygalae Radix and water-boiled Polygalae Radix were determined by UPLC-triple quadrupole tandem mass spectrometry（UPLC-QQQ-MS/MS）. ResultDuring the boiling process with licorice juice and water， onjisaponin B could be hydrolyzed to produce 4-methoxycinnamic acid， desacylsenegin Ⅲ， polygalasaponin ⅩⅩⅧ and tenuifolin， onjisaponin Z could be hydrolyzed to produce 3，4，5-trimethoxycinnamic acid， onjisaponin TF， polygalasaponin ⅩⅩⅧ and tenuifolin， onjisaponin F could be hydrolyzed to produce 3，4，5-trimethoxycinnamic acid， onjisaponin G， polygalasaponin ⅩⅩⅧ and tenuifolin， and polygalasaponin ⅩⅩⅧ was hydrolyzed to produce tenuifolin. After being boiled with licorice juice or water， the content of onjisaponin B decreased significantly（P<0.05， P<0.01）， but the contents of onjisaponin Z， onjisaponin F， polygalasaponin ⅩⅩⅧ and tenuifolin increased significantly（P<0.05， P<0.01） in Polygalae Radix. Compared with the water-boiled products， the contents of onjisaponin Z and tenuifolin increased significantly（P<0.05， P<0.01）， and the change of tenuifolin content was the most significant in the licorice-boiled products.However， there was no significant difference in the content of onjisaponin B， onjisaponin F and polygalasaponin ⅩⅩⅧ between the water-boiled products and the licorice-boiled products. ConclusionBeing boiled with licorice juice or water can hydrolyze onjisaponin B， onjisaponin Z， onjisaponin F and polygalasaponin ⅩⅩⅧ， and generate secondary glycosides and aglycones（organic acids） through deglycosylation， which leads to obvious changes in the contents of onjisaponins after Polygalae Radix being processed.It is inferred that licorice juice can promote the hydrolysis of some onjisaponins in Polygalae Radix to onjisaponin Z and tenuifolin.This study provides an experimental basis for revealing processing mechanism of Polygalae Radix.

Discovery of pulmonary fibrosis inhibitor targeting TGF-β RI in Polygonum cuspidatum by high resolution mass spectrometry with in silico strategy / 药物分析学报

Pulmonary fibrosis(PF)is an irreversible lung disease that is characterized by excessive scar tissue with a poor median survival rate of 2-3 years.The inhibition of transforming growth factor-β receptor type-Ⅰ(TGF-β RI)by an appropriate drug may provide a promising strategy for the treatment of this disease.Polygonum cuspidatum(PC)is a well-known traditional Chinese herbal medicine which has an anti-PF effect.Accordingly,a combination of high resolution mass spectrometry with an in silico strategy was developed as a new method to search for potential chemical ingredients of PC that target the TGF-β RI.Based on this strategy,a total of 24 ingredients were identified.Then,absorption,distribution,meta-bolism,and excretion(ADME)-related properties were subsequently predicted to exclude compounds with potentially undesirable pharmacokinetics behaviour.Molecular docking studies on TGF-β RI were adopted to discover new PF inhibitors.Eventually,a compound that exists in PC known as resveratrol was proven to have excellent biological activity on TGF-β RI,with an ICso of 2.211 μM in vitro.Furthermore,the complex formed through molecular docking was tested via molecular dynamics simulations,which revealed that resveratrol had strong interactions with residues of TGF-β RI.This study revealed that resveratrol has significant potential as a treatment for PF due to its ability to target TGF-β RI.In addition,this research demonstrated the exploration of natural products with excellent biological activities toward specific targets via high resolution mass spectrometry in combination with in silico technology is a promising strategy for the discovery of novel drugs.

High-throughput screening of drug residues in aquatic products by liquid chromatography-electrostatic field track well high resolution mass spectrometry / 上海预防医学

ObjectiveIn order to ensure the food safety of cultured aquatic products in China， strengthen the supervision and improve detection efficiency， a high-throughput screening method for drug residues in aquatic products by mass spectrometry is to be developed. MethodsLiquid chromatography-electrostatic field track well high resolution mass spectrometry and C18 column were used to separate and collect the information of ion fragments and retention time of 195 veterinary drug standards in positive ion mode， and drug residues in aquatic products were extracted by organic solvents. ResultsThe mass spectrum database and detection methods of 195 veterinary drug standards were established， and a pre-treatment method was developed for extracting veterinary drug residues from aquatic products by acetonitrile and ethyl acetate. The detection method was sensitive， accurate， simple and rapid， and the adding standard recovery of more than 80% compounds reached 70%‒110%. ConclusionThe establishment of this method can quickly and massively screen drug residues in aquatic products， provide technical support for food safety risk monitoring and supervision， cope with the increasing demand for aquatic products and aquaculture volume year by year， and provide a theoretical basis for the subsequent development of mass spectrometry high-throughput detection methods.

Mechanism Analysis of Strengthening Anti-cerebral Ischemia Injury of Chuanxiong Rhizoma Processed with Wine Based on UHPLC-Q-Orbitrap HRMS and Integrated Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo analyze the differential components in water extract of Chuanxiong Rhizoma before and after processing with wine， and to explore the molecular mechanism of Chuanxiong Rhizoma processed with wine in enhancing anti-cerebral ischemia injury. MethodUltra high performance liquid chromatography tandem quadrupole orbitrap high resolution mass spectrometry （UHPLC-Q-Orbitrap HRMS） was used to qualitatively analyze the main chemical components in water extract of Chuanxiong Rhizoma based on the spectral information of compound， comparison of reference substance and references. The chemical pattern recognition method was used to screen the differential components of Chuanxiong Rhizoma before and after processing. Based on these differential components， the potential targets of differential components were predicted by online databases， and the related targets of cerebral ischemia were searched. Cytoscape 3.6.0 was used to establish the network diagram of differential components-action targets-diseases of Chuanxiong Rhizoma processed with wine. The protein-protein interaction （PPI） network of intersection targets was constructed by STRING 11.5. The potential targets of differential components against cerebral ischemia were analyzed by Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analysis through DAVID 6.8. At the same time， the chemical compounds with high relative content and increased peak area after wine processing were docked with their corresponding targets to verify the mechanism of enhanced effect after wine processing. ResultA total of 71 chemical components were identified from Chuanxiong Rhizoma， 34 differential components and 603 potential targets were screened out. At the same time， a total of 769 disease targets and 60 intersection targets were obtained. Seven key targets were identified through PPI network analysis， including JUN， signal transducer and activator of transcription 3 （STAT3）， mitogen-activated protein kinase 3 （MAPK3）， interleukin-1β （IL-1β）， vascular endothelial growth factor A （VEGFA）， Caspase-3 （CASP3） and mtrix metalloproteinase 9 （MMP9）. Tumor necrosis factor （TNF） signaling pathway was the main differential signaling pathway. The results of molecular docking showed that differential components （senkyunolide K， senkyunolide F， 3-n-butylphthalide， Z，Z′-6，8′，7，3′-diligustilide， ferulic acid and Z-ligustilide） and corresponding targets had good binding activities. ConclusionThe synergistic mechanism of Chuanxiong Rhizoma processed with wine may be related to the enhanced inhibitory effect of inflammatory reaction.

Serum Metabonomics Analysis of Diarrhea Mice Intervened by Zingiberis Rhizoma Recens Based on UPLC-QE-Orbitrap-MS / 中国实验方剂学杂志

ObjectiveTo investigate the antidiarrheal effect and mechanism of Zingiberis Rhizoma Recens on diarrhea mice， and to provide research basis for the inhibition of intestinal peristalsis by Zingiberis Rhizoma Recens and its application in the treatment of gastrointestinal diseases. MethodThe diarrhea model of mice was established by Sennae Folium. The control group， model group， Zingiberis Rhizoma Recens low-， medium-， high-dose groups （0.1， 0.32， 1.0 g·kg-1） and loperamide group （1.6 g·kg-1） were set. The intervention effect of Zingiberis Rhizoma Recens with different doses on diarrhea mice was detected by diarrhea score， incidence rate of loose stools （LSIR）， grade of average loose stools （ALSG）， diarrhea index （DI）， intestinal propulsion rate and intestinal pathological section. The serum metabonomics of mice was analyzed by ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA）. The conditions were as follows：mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B）， flow rate of 0.4 mL·min-1， injection volume of 5 µL， electrospray ionization （ESI）， positive and negative ion detection modes， acquisition range of m/z 100-1 500. ResultCompared with the model group， Zingiberis Rhizoma Recens high-dose group could obviously reduce the diarrhea score， LSIR， ALSG， DI and intestinal propulsion rate （P<0.05， P<0.01）， and improve the intestinal mucosal injury. There were 40 main differential metabolites among the control group， model group and Zingiberis Rhizoma Recens high-dose group， including glucose 1-phosphate， xanthine， xanthosine and so on. The metabolic pathways mainly included starch and sucrose metabolism， amino sugar and nucleotide sugar metabolism， fructose and mannose metabolism， tryptophan metabolism， and galactose metabolism. ConclusionZingiberis Rhizoma Recens can inhibit intestinal peristalsis in diarrhea mice and exert antidiarrhoea effect， the mechanism of which may be related to the regulation of carbohydrate and amino acid metabolism.

Analysis of Five Mushroom Toxins in Blood by UPLC-HRMS / 法医学杂志

OBJECTIVES@#To develop a method for the simultaneous and rapid detection of five mushroom toxins (α-amanitin, phallacidin, muscimol, muscarine and psilocin) in blood by ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS).@*METHODS@#The blood samples were precipitated with acetonitrile-water solution(Vacetonitril∶Vwater=3∶1) and PAX powder, then separated on ACQUITY Premier C18 column, eluted gradient. Five kinds of mushroom toxins were monitored by FullMS-ddMS2/positive ion scanning mode, and qualitative and quantitative analysis was conducted according to the accurate mass numbers of primary and secondary fragment ions.@*RESULTS@#All the five mushroom toxins had good linearity in their linear range, with a determination coefficient (R2)≥0.99. The detection limit was 0.2-20 ng/mL. The ration limit was 0.5-50 ng/mL. The recoveries of low, medium and high additive levels were 89.6%-101.4%, the relative standard deviation was 1.7%-6.7%, the accuracy was 90.4%-101.3%, the intra-day precision was 0.6%-9.0%, the daytime precision was 1.7%-6.3%, and the matrix effect was 42.2%-129.8%.@*CONCLUSIONS@#The method is simple, rapid, high recovery rate, and could be used for rapid and accurate qualitative screening and quantitative analysis of various mushroom toxins in biological samples at the same time, so as to provide basis for the identification of mushroom poisoning events.

Analysis on Chemical Constituents in Rhizomes of Bergenia scopulosa by UHPLC-Q Exactive Focus MS/MS / 中国实验方剂学杂志

Objective:Ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry （UHPLC-Q Exactive Focus MS/MS） was developed to rapidly analyze and identify the chemical components in the rhizomes of <italic>Bergenia scopulosa</italic>. Method:The 75% methanol extract of <italic>B</italic>.<italic> scopulosa</italic> rhizomes was analyzed on a Thermo Accucore aQ RP18 column （2.1 mm×150 mm， 2.6 μm） with the mobile phase of methanol （A） and 0.1% formic acid aqueous solution （B） for gradient elution （0-40 min 5%-95%A， 40-45 min 95%A）， the flow rate was 0.3 mL·min^-1 and the column temperature was at 30 ℃. The information of the chemical constituents was acquired in positive and negative ion modes by heated electrospray ion source （HESI）， and the scanning range was <italic>m</italic>/<italic>z</italic> 80-1 200. Result:A total of 66 chemical constituents were identified， including 2 free amino acids， 7 bergenin derivatives， 15 flavonoids， 15 organic acids， 25 glycosides， and 2 others. Conclusion:The chemical constituents in the rhizomes of <italic>B</italic>.<italic> scopulosa</italic> can be identified systematically， accurately and rapidly by this method. Among them， 8 compounds were unambiguously identified by comparing with reference substances （succinic acid， arbutin， gallic acid， protocatechuic acid， bengenin， catechin， chlorogenic acid and caffeic acid）， 51 compounds were found from <italic>B</italic>.<italic> scopulosa</italic> for the first time and 28 compounds were found from the genus <italic>Bergenia</italic> for the first time. This paper can provide an important basis for the further material basis clarification and quality assessment of <italic>B</italic>.<italic> scopulosa</italic>.

Analysis and Identification of Flavonoids in Citri Reticulatae Pericarpium with Different Aging Time by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective:To analyze and identify the flavonoids of Citri Reticulatae Pericarpium with different aging time by an ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS）. Method:Compounds were separated on Agilent Extend-C₁₈ column （3.0 mm×100 mm， 1.8 μm）， mobile phase was 0.1% acetic acid aqueous solution （A）-0.1% acetic acid methanol solution （B） for gradient elution （0-25 min， 5%-95%B； 25-30 min， 95%B； 30-30.1 min， 95%-5%B； 30.1-35 min， 5%B）， the flow rate was 0.4 mL·min^-1， and the column temperature was set at 30 ℃. High resolution mass spectrometry was performed with electrospray ionization （ESI）， and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS²）. The multistage ion fragment information combined with mzCloud network database， local high resolution mass spectrometry database of traditional Chinese medicine components （OTCML）， literature information and relevant reference materials were used for accurate qualitative analysis. Result:Totally 43 flavonoids in Citri Reticulatae Pericarpium were identified， including 24 flavones， 5 flavonols， 13 dihydroflavones and 1 chalcone. The flavonoids in samples with different aging time were basically consistent in material types， but the peak area was different. According to the comparison of relative content in the peak area， it was found that the relative contents of 30 flavonoids showed an overall increasing trend with the increase of aging time. Among them， the relative contents of 24 flavonoids （such as hesperidin， diosmin， 6-demethoxytangeretin， nobiletin and tangeretin） increased significantly. There was no significant change in the relative contents of the other 13 flavonoids （such as naringenin and neohesperidin）. Conclusion:An efficient method is established in this paper to identify flavonoids in Citri Reticulatae Pericarpium with different aging time and their relative content changes rapidly and accurately. The findings provide a methodological reference for the study on pharmacodynamic material base and quality control of Citri Reticulatae Pericarpium， and it provides experimental basis that drugs processed long time ago have better effect of Citri Reticulatae Pericarpium.

Identification of Chemical Constituents in Coptis chinensis Inflorescence by UPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

Objective:An ultra-high performance liquid chromatography coupled with quadrupole-orbitrap high resolution mass spectrometry （UPLC-Q-Orbitrap HRMS） was developed to analyze and identify the chemical constituents in <italic>Coptis chinensis</italic> inflorescence. Method:The chromatographic separation was performed on ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.7 μm） with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-15 min， 10%-22%B； 15-20 min， 22%B； 20-25 min， 22%-44%B； 25-35 min， 44%-50%B； 35-40 min， 50%-60%B； 40-55 min， 60%-85%B）， the flow rate was 0.15 mL·min^-1， the injection volume was 3 μL and the column temperature was 30 ℃. HRMS was equipped with electrospray ionization （ESI） and scanned in positive and negative ion modes by means of full scan/data dependent secondary scan （Full MS/dd-MS²）. Compound Discoverer 3.0 software combined with mzCloud， mzVault， ChemSpider databases and HRMS database of components in traditional Chinese medicine were used to analyze and identify the collected data by HRMS， based on accurate relative molecular mass， retention time and characteristic ion fragmentation of the compounds， as well as literature information and relevant reference materials. Result:A total of 51 chemical constituents were identified in <italic>C</italic>.<italic> chinensis</italic> inflorescence， including 16 alkaloids， 14 flavonoids， 7 phenylpropanoids， 7 organic acids and 7 others. Among them， 10 components ［berberine， palmatine， coptidine， rutin， quercetin， isoquercitrin， chlorogenic acid， cryptochlorogenic acid，<italic> D</italic>-（-） quinic acid and <italic>D</italic>-proline］ were unambiguously identified by comparing with reference standards. Conclusion:The established UPLC-Q-Orbitrap HRMS can be used to accurately analyze and identify chemical constituents of <italic>C. chinensis</italic> inflorescence. A total of 41 chemical constituents are reported from <italic>C. chinensis</italic> inflorescence for the first time and 6 alkaloids are found from the <italic>C. chinensis</italic> for the first time. These findings can provide methodological reference and experimental basis for the basic research of quality evaluation and efficacy materials of <italic>C. chinensis</italic> inflorescence， and lay a foundation for its further development and utilization.

Analysis of Chemical Constituents in Microctis Folium by UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To analyze the chemical constituents in Microctis Folium by ultra performance liquid chromatography-quadrupole-time-of-flight high resolution mass spectrometry （UPLC-Q-TOF-MS/MS）. Method:Waters CORTECS UPLC C₁₈ column （2.1 mm×150 mm， 1.6 μm） was used for chromatographic separation with the mobile phase of methanol （A） -0.1% formic acid solution （B） for gradient elution （0-4 min， 14%-30%A； 4-16 min， 30%-58%A； 16-25 min， 58%-78%A； 25-25.1 min， 78%-98%A； 25.1-29 min， 98%A）， the flow rate was 0.25 mL· min^-1， the injection volume was 1 μL. The electrospray ionization （ESI） was adopted for determining the chromatographic effluent under positive and negative ion modes， the main chromatographic peaks were assigned and distinguished by Q-TOF， and the scanning range was <italic>m</italic>/<italic>z</italic> 100-1 500. Result:A total of 31 chemical constituents in Microctis Folium were identified by confirmation of reference substances， literature comparison and high resolution mass spectrometry data analysis. The chemical constituent cluster was composed of 28 flavonoids （9 flavone C-glycosides， 10 flavonols and their glycosides， 8 proanthocyanidins， 1 xanthone） and 3 organic acids （caffeic acid， <italic>p</italic>-coumaric acid， ferulic acid）. Conclusion:UPLC-Q-TOF-MS/MS technique provides a simple， rapid and accurate method for the identification of chemical constituents in Microctis Folium. Flavone C-glycosides， flavonol oxyglycosides and proanthocyanidins are the main chemical constituents. The 7 proanthocyanidins are reported for the first time in this herb. In conclusion， the chemical profile of Microctis Folium is characterized and the findings are meaningful for the in-depth quality assessment and material basis clarification of Microctis Folium.

Analysis of Chemical Constituents in Qizhi Jiangtang Capsules Based on UPLC-QE-Orbitrap-MS / 中国实验方剂学杂志

Objective:To systematically analyze the chemical constituents of Qizhi Jiangtang capsules by ultra performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry （UPLC-QE-Orbitrap-MS）. Method:Analysis was conducted on a ACQUITY UPLC HSS T3 column （2.1 mm×100 mm， 1.8 μm） with acetonitrile （A）-water （B） as the mobile phase for gradient elution （0-13 min， 1%-25%A； 13-21 min， 25%-35%A； 21-28 min， 35%-85%A； 28-30 min， 85%-100%A； 30-32 min， 100%-1%A）. The flow rate was 0.2 mL·min^-1， the column temperature was 30 ℃， and the volume of sample injection was 3 μL. Electrospray ionization （ESI） was used to collect data in the negative and positive ion modes with the scanning range of <italic>m</italic>/<italic>z</italic> 100-1 500. Meanwhile， a variety of MS analytic methods were used， including comparing with the information of control substances， self-built compounds database and literature references， diagnostic ion filtering， Compound Discoverer 3.0 software， for identification of the chemical components. Result:Based on the above strategy， a total of 52 compounds were identified in Qizhi Jiangtang capsules， and the sources of these compounds were identified. Amino acids were mainly derived from Hirudo， phenylpropanoids were derived from Astragali Radix and Rehmanniae Radix， iridoid glycosides were derived from Rehmanniae Radix， coumarins and triterpenes were derived from Astragali Radix， flavonoids were from Astragali Radix and Polygonati Rhizoma. Conclusion:The established UPLC-QE-Orbitrap-MS analytical method can comprehensively and rapidly analyze and identify of the chemical constituents in Qizhi Jiangtang capsules. Many of the ingredients have been proved by modern pharmacological studies to have the effect of improving related symptoms of diabetes and its complications， reflecting the characteristics of synergistic action of multiple components in Qizhi Jiangtang capsules. This study can provide reference for the further research on the pharmacodynamic material basis and the quality control of Qizhi Jiangtang capsules.