Application of an Organophosphorus Compound-DEPBT as Coupling Reagent in Liquid Phase Peptide Synthesis

DEPBT, a novel coupling reagent containing phosphorus, was used in synthesis of pentapeptide Boc-Ile-Asn-Met-Trp(For)-Gly.OMe by solution method.

Effect of pediatric tuina on hypothalamic metabolites in young rabbits using liquid chromatography-mass spectrometry

Objective: To investigate the changes in the hypothalamic metabolites of lipopolysaccharide(LPS) febrile young rabbits after the treatment with pediatric tuina.Methods: A total of 30 young rabbits were randomly assigned into three groups: the normal group, the model group, and the tuina group. Both the model group and the tuina group were injected intravenously with LPS. “Six antipyretic manipulations”(pushing Tianmen, pushing Kangong, kneading Taiyang,kneading Erhougaogu, clearing Tianheshui, and pushing Jizhu) were administered 1 h after the LPS injection in the tuina group. The rectal temperatures of the young rabbits were monitored during the experiment to explore the antipyretic effect. Three hours after the injection, the content of interleukin-1β(IL-1β), tumor necrosis factor(TNF)-a, and prostaglandin E;(PGE;) in the serum was detected. In addition, liquid chromatography-mass spectrometry(LC-MS) was used for the hypothalamus metabolomics.Results: Compared with the model group, the rectal temperature of the tuina group was decreased at 2 h and 3 h after the LPS injection(P =.04, P =.03, respectively), and the content of IL-1β, TNF-a, and PGE2was decreased(P =.03, P =.003, and P =.008, respectively). The metabolomics results showed that there were 23 potential biomarkers after the tuina intervention, enriching 27 pathways. Lipid metabolites,especially glycerophospholipids, were a majority of the altered metabolites. The primary metabolic pathways affected by tuina included the arachidonic acid metabolism, the GABAergic synapse, D-glutamine and D-glutamate metabolism, and the glutamatergic synapse.Conclusion: Pediatric tuina reduced the temperature of the febrile rabbits and downregulated the expression of IL-1β, TNF-a, and PGE2, and the antipyretic mechanism may be related to changes in metabolites and metabolic pathways in the hypothalamus.

Rapid Screening of 26 Organophosphorus Pesticides in Fresh Milk by Ultra Liquid Chromatography Coupled With Quadrupole-Time of Flight Mass Spectrometry

[Objectives]A rapid screening and analysis method for 26 organophosphorus agrochemicals in fresh milk was established using ultra performance liquid chromatography coupled with quadrupole-time of flight mass spectrometry.[Methods]Raw milk was extracted with acetonitrile solution containing 0.2%formic acid by volume,and purified with a Dikma ProElut QuECHERS solid phase extraction cartridge.Target compounds were separated on a Waters ACQUITY UPLC HSS T3 chromatographic column(2.1 mm×50 mm,1.8μm)with methanol-water solution as a mobile phase for gradient elution,and through scanning with an electrospray ion source in positive ion mode,26 kinds of organophosphorus agrochemicals could be accurately qualitatively determined within 10 min.[Results]When using formic acid acetonitrile with a volume fraction of 0.2%,there were more types of detected compounds and a greater recovery;and using B cartridge could effectively eliminate the interference of non-polar substances such as phospholipids,achieve higher number of detected compounds than those of A and C,and well separate the 26 kinds of agrochemical residues.[Conclusions]This study provides a reference method for the rapid screening of agrochemical residues in dairy cows in the future.

Lysophosphatidylcholine Biomarkers of Lung Cancer Detected by Ultra-performance Liquid Chromatography Coupled with Quadrupole Time-of-flight Mass Spectrometry

Centrifugal ultrafiltration after methanol extraction of whole plasma was used as an optimal condition for the preparation of blood plasma before metabonomic studies. The plasma samples from 102 lung cancer patients and 34 healthy volunteers were prepared with this approach. With ultra-performance liquid chromatography（UPLC） coupled with quadrupole time-of-flight mass spectrometry（Q-TOF MS） analysis, the samples were investigated in order to find potential disease biomarkers. After data acquisition, orthogonal signal correction partial least squares models were built to differentiate the healthy volunteers from lung cancer patients and to identify metabolites that showed significantly different expression between the two groups. Several metabolite ions were identified as potential biomarkers according to the variable importance in the project（VIP） value in both ion modes. Five lysophosphatidylcholines were further identified as specifically lysoPC 16：0, isomer of lysoPC 16：0, lysoPC 18：0, lysoPC 18：1 and lysoPC 18：2. These results suggest that UPLC coupled with Q-TOF MS is an effective technique for the analysis of plasma metabolites in metabonomic studies.

Analysis of Low-polar Ginsenosides in Steamed Panax Ginseng at High-temperature by HPLC-ESI-MS/MS

A high performance liquid chromatography coupled with electrospray ionization-tandem mass spectrome try（HPLC-ESI-MS/MS） method was developed for the analysis and identification of ginsenosides in the extracts of raw Panax ginseng（RPG） and steamed Panax ginseng at high temperatures（SPGHT）. A total of 25 ginsenosides were extracted include of which 10 low-polar ginsenosides, such as ginsenosides F4, Rk3, Rh4, 20S-Rg3, 20R-Rg3 and so on, were identified according to their HPLC retention time and MS/MS data. The results indicated that the low polar ginsenosides were seldom found in RPG. For the exploration of the transformation pattern of the ginsenosides in steam processing, the standards of ginsenosides Re, Rg1, Rb1, Rc, Rb2, Rb3 and Rd were selected and hydrolyzed at a temperature of 120 oC. The results show that these polar ginsenosides can be converted to low-polar ginsenosides such as Rg2, Rg6, F4, Rk3 and Rg5 by hydrolyzing the sugar chains.

liver and the defects of cholesterol and bile acids biosynthesis: rare disorders many diagnostic pitfalls

In recent decades, biotechnology produced a growth of knowledge on the causes and mechanisms of metabolic diseases that have formed the basis for their study, diagnosis and treatment. Unfortunately, it is well known that the clinical features of metabolic diseases can manifest themselves with very different characteristics and escape early detection. Also, it is well known that the prognosis of many metabolic diseases is excellent if diagnosed and treated early. In this editorial we briefly summarized two groups of inherited metabolic diseases, the defects of cholesterol biosynthesis and those of bile acids. Both groups show variable clinical manifestations but some clinical signs and symptoms are common in both the defects of cholesterol and bile acids. The differential diagnosis can be made analyzing sterol profiles in blood and/or bile acids in blood and urine by chromatographic techniques(GC-MS and LC-MS/MS). Several defects of both biosynthetic pathways are treatable so early diagnosis is crucial. Unfortunately their diagnosis is made too late, due either to the clinical heterogeneity of the syndromes(severe, mild and very mild) that to the scarcity of scientific dissemination of these rare diseases. Therefore, the delay in diagnosis leads the patient to the medical observation when the disease has produced irreversible damages to the body. Here, we highlighted simple clinical and laboratory descriptions that can potentially make you to suspect a defect in cholesterol biosynthesis and/or bile acids, as well, we suggest appropriate request of the laboratory tests that along with common clinical features can help to diagnose these defects.

Identification and determination of the major constituents in traditional Chinese medicine Longdan Xiegan Pill by HPLC-DAD-ESI-MS

A novel and sensitive HPLC-UV method has been developed for the simultaneous determination of twelve major compounds in Longdan Xiegan Pill.The chemical profile of the twelve compounds,including geniposidic acid（1）,geniposide（2）,gentiopicroside（3）,liquiritin（4）,crocin（5）,baicalin（6）,wogonoside（7）,baicalein（8）,glycyrrhizic acid（9）,wogonin（10）,oroxylin A（11）and aristolochic acid A（12）,was acquired using high-performance liquid chromatography-diode array detector coupled with an electrospray tandem mass spectrometer（HPLC-DAD-ESI-MS）.The analysis was performed on a Dikma Platisil ODS C18 column（250 mm×4.6 mm,5 μm）with a gradient solvent system of acetonitrile-0.1% aqueous formic acid.The validation was carried out and the linearities（r〉0.9996）,repeatability（RSD〈1.8%）,intra-and inter-day precision（RSD〈1.3%）,and recoveries（ranging from 96.6% to 103.4%）were acceptable.The limits of detection（LOD）of these compounds ranged from 0.29 to 4.17 ng.Aristolochic acid A,which is the toxic ingredient,was not detected in all the batches of Longdan Xiegan Pill.Furthermore,hierarchical cluster analysis was used to evaluate the variation of the herbal prescription.The proposed method is simple,effective and suitable for the quality control of this traditional Chinese medicine（TCM）.

Combined Use of NMR, LC-ESI-MS and Antifungal Tests for Rapid Detection of Bioactive Lipopeptides Produced by Bacillus

The science and technology interact with the art in several ways. Biotechnological coupled with analytical approaches can play an important role in protecting and preserving cultural heritage for future generations. Many microorganisms influenced by environmental conditions are the main responsible for biological contamination in built heritage. Biocides based on chemical compounds have been used to mitigate this problem. Thus, it is vitally important to develop proper remediation actions based on environmentally innocuous alternative. Bacillus specie is emerging as an optimistic alternative for built heritage treatment due to their capacity to produce secondary metabolites with antagonistic activities against many fungal pathogens. Therefore, the intent of this work was to access a rapid evaluation of antifungal potential of bioactive metabolites produced by Bacillus strains and simultaneously their characterization using spectroscopic (NMR) and chromatographic techniques (LC-ESI-MS). The high antifungal activity obtained for Bacillus sp. active compounds produced in this study confirms the great potential to suppress biodeteriogenic fungi growth on historical artworks. Additionally, the proposed methodology allowed to access bioactive metabolites produced without need of the laborious total previous isolation and could be used as a viable alternative to be employed for screening and production of new green biocides.

Evaluating the Compatibility Mechanism of Shengxian Decoction Based on an Excretion Study of 18 Bioactive Constituents in Rat Biosamples

Introduction:The Shengxian decoction(SXT),a Chinese herbal medication used to treat heart failure,is composed of Astragali Radix,Anemarrhenae Rhizoma,Bupleuri Radix,Cimicifuage Rhizoma,and Platycodonis Radix.Knowledge of the excretion of the active compounds in this herbal medication is vital for investigating the underlying mechanisms behind its compatibility.However,this remains unclear.Methods:Liquid chromatography coupled with mass spectrometry was performed in both positive and negative modes to assay 18 constituents of SXT from rat urinary,fecal,and biliary samples.Results:The methodology was validated and showed good linearity(r≥0.9),precision,stability,repeatability,and recovery.The relative standard deviations and relative errors for the precision and accuracy did not exceed 15%.The recoveries of all the compounds ranged from 77.37%to 114.82%,and the matrix effect was between 82.53%and 105.71%.The results suggested that,when combined with Platycodonis Radix,the levels of 14 constituents from the four herbs were reduced in the urine,while the levels of six constituents were reduced in the feces.Conclusions:The comparative excretion results indicated that Platycodonis Radix reduced the excretion of compounds in SXT,demonstrating the compatibility mechanism and holistic properties of these compounds,favoring the pharmacological effects of SXT.

A rapid and simple ultraperformance liquid chromatography coupled to tandem mass spectrometry(UPLC-MS/MS) method for the detection of glucuronic acid-conjugated steroid metabolites

In the present study, we effectively detected 10 steroids and glucuronic acid-conjugated steroid metabolites in 12 min by ultraperformance liquid chromatography coupled to tandem mass spectrometry （UPLC-MS/MS）. Steroids testosterone （T）, 5ct-dihydrotestosterone （DHT）, androsterone （ADT）, etiocholanolone （ETIO）, estradiol （E2） and their glucuronide conjugates were well-separated on an Eclipse Plus C18 column （2.1 mm×50 ram, RRHD 1.8μm）. The mobile phase consisted of a mixture of methanol and ultrapure water （containing I mM ammonium formate） at a ratio of 60：40 （v/v）, and the flow rate was set at 0.25 mL/min. The LC eluate was detected by electrospray ionization （ESI） source in both positive and negative ion modes. Neutral loss （NL of 176, 194, 211 and 229 Da in positive mode） and precursor ion （PI ofm/z 141,159 and 177 in positive mode and 75, 85 and 133 in negative mode） methods were applied for the detection of steroid glucuronides. The multiple reaction monitoring （MRM） transitions were m/z 289.3→97.1,291.3→105, 291.3→199.2, 273.2→145.4 and 255.2→159.1 for T, DHT, ADT, ETIO and E2 in positive mode, respectively; as well as m/z 463.3→85 for T glucuronide （T-G）, m/z 465.3→75 for DHT glucuronide （DHT-G）, ADT glucuronide （ADT-G）, ETIO glucuronide （ETIO-G） and m/z 447.3→271 for E2 glucuronide （Ez-G） in negative mode. In addition, the analytical method was also applied for the detection of steroid glucuronides in pooled human liver microsomes （HLM）, which might serve as a basis for further investigation of steroid metabolism in vivo and in vitro.

Separation and identification of moxifloxacin impurities in drug substance by high-performance liquid chromatography coupled with ultraviolet detection and Fourier transform ion cyclotron resonance mass spectrometry

In this paper, a high-performance liquid chromatography coupled with ultraviolet detection and Fourier transform-ion cyclotron resonance mass spectrometry （HPLC-UV/FrICRMS） method was described for the investigation of impurity profile in moxifloxacin （MOX） drug substance and chemical reference substance. Ten impurities were detected by HPLC-UV, while eight impurities were identified by using the high accurate molecular mass combined with multiple-stage mass spectrometric data and fragmentation rules. In addition, to our knowledge, five impurities were founded for the first time in MOX drug substance.

COMPREHENSIVE TWO-DIMENSIONAL LIQUID CHROMATOGRAPHY COUPLED WITH QUADRUPLE TIME-OF-FLIGHT MASS SPECTROMETRY FOR CHEMICAL CONSTITUENTS ANALYSIS OF TRIPTERYGIUM GLYCOSIDES TABLETS

Comprehensive two-dimensional liquid chromatography platform(LC×LC)coupled with quadrupole time-of-flight(QTOF)mass spectrometry(MS)is developed to separate,identify and relatively determine the chemical constituents of two types of tripterygium glycosides tablets(TGT).The types and relative contents of the constituents discovered in two kinds of TGT tablets were subsequently compared.C8andC18 column were used for the separation of the first

Pharmacokinetic Comparison of Four Major Bio-Active Components of Naoxintong Capsule in Normal and Acute Blood Stasis Rats Using Ultra-Performance Liquid Chromatography Coupled with Triple-Quadrupole Mass Spectrometry

Objective: To compare the pharmacokinetic differences of the main components of Naoxintong capsule(NXTC) in normal and acute blood stasis rats. Materials and Methods: Rats were subcutaneously injected with adrenaline hydrochloride twice;during the two subcutaneous injections, the rats were placed in ice water for 4 min to reproduce the model rat of acute blood stasis. The normal and acute blood stasis rats were administrated a 5.04 g/kg dose of NXTC suspension. Then, blood samples were collected from the posterior retinal venous plexus at different time points. Plasma concentrations of four major bio-active components including caffeic acid, ferulic acid, formononetin, and tanshinone IIA in NXTC were measured using ultra-performance liquid chromatography coupled with triple-quadrupole mass spectrometry. Phoenix Win Nonlin v6.2 software was used to calculate the pharmacokinetic parameters. Results: Compared with the normal rats, the acute blood stasis rats showed a significant decrease in C_(max) of ferulic acid and formononetin, AUC_(all) of caffeic acid and ferulic acid, and AUC_(INF_obs) of ferulic acid. Conversely, an increase in the Vz_F_obs and MRT_(last) of ferulic acid and caffeic acid was observed. These findings demonstrate that the absorption of the four NXTC components was weakened in the acute blood stasis rats and that the elimination time was prolonged. Conclusions: The significant difference in some parameters of the four NXTC components between the normal and acute blood stasis rats might be caused by an increase in blood viscosity and the subsequent slowing down of blood flow in the acute blood stasis rats. The pharmacokinetic study conducted in pathological state can provide important information and scientific basis for further rational clinical application of NXTC.

Rapid Differentiation of Aconiti Kusnezoffii Radix from Different Geographic Origins Using Ultra-Performance Liquid Chromatography Coupled with Time-of-Flight Mass Spectrometry

Objective:There are different geographic origins of Aconiti Kusnezoffii Radixs(AKRs)sold in the market with different quality.This study aims to establish a rapid analysis method to distinguish the different geographic origins of AKRs and to realize the rapid evaluation of their quality.Methods:An ultra-performance liquid chromatography coupled with time-of-flight mass spectrometry(UPLC-Q-TOF MS)method was utilized to acquire the constituents'information of AKRs from different geographic origins.MSE data and Progenesis QI software were employed to identify the chemical constitutes.Principal component analysis(PCA)was applied to comparing MS data to find the chemical markers of AKRs from different geographic origins.Results:Twenty-three components were detected and 17 out of them were identified,including diester-diterpenoid alkaloids,monoester-diterpenoid alkaloids,and amine-diterpenoid alkaloids.Three pairs of isomers were detected and two of them were distinguished by the retention time of standard samples.Thirteen chemical markers were screened out through PCA and orthogonal partial least square discriminant analysis.Through detecting Napelline or isomer of Napelline(m/z 360.2530)and Aconifine(m/z 662.3170),AKRs from inner Mongolia autonomous could be screened.According to the existence of benzoylaconine(m/z 604.3108)and Indaconitine(m/z 630.3159),it could be confirmed that the AKRs are from Xinjiang Uygur autonomous.AKRs that cannot detect compounds above-mentioned could be from Liaoning or Shanxi Province.Conclusions:The chemical profile could be used not only to distinguish the AKRs from different geographic origins but also to identify the true and false of AKRs.This study lays a foundation for the study of efficacy and toxic of AKRs.

Rapid Determination of Tetrodotoxin in Human Plasma by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry

A sensitive analytical method was developed to determine tetrodotoxin（TTX） in human plasma samples using protein precipitation, followed by ultra performance liquid chromatography（UPLC） analysis coupled with tandem mass spectrometry（MS/MS） using 11-deoxytetrodotoxin（11-deoxyTTX） as an internal standard. The plasma samples were prepared using protein precipitation prior to being analyzed by UPLC-MSfMS to identify TTX over a zwitterionic-hydrophilic interaction liquid chromatography column. The retention time values of TTX and 11-deoxyTTX were 4.12 and 3.67 min, respectively. TTX and 11-deoxyTTX were monitored and quantitated on the basis of their ion transitions for their respective precursor ions to their product ions（i.e., m/z 320.0→162. l for TTX and m/z 304.0→176.0 for 11-deoxyTTX） in the multiple reaction-monitoring mode. The lower limit of quantification of this method was determined to be 0.0199 ug/mL. This method showed good linearity for plasma samples that contained TTX concentrations in the range of 0.0199--1.99 ng/mL. The specificity, precision, accuracy, matrix effect, and stability characteristics of this method were also examined. The intra-assay precision and accuracy ranged from 1.89% to 6.00% and from 92.21% to 100.00%, whereas the inter-assay precision and accuracy ranged from 0.64% to 7.75% and from 99.38% to 101.26%, respectively. This new method therefore represents a rapid, accurate, reliable, and highly sensitive method for the qualitative and quantitative analyses of a trace amount of TTX in human plasma samples.

GLOBAL ANALYSIS OF THE METABOLITES OF CURCUMIN IN RATS BY ULTRA PERFORMANCE LIQUID CHROMATOGRAPHY COUPLED WITH QUADRUPOLE TIME OF-FLIGHT TANDEM MASS SPECTROMETRY

Curcumin,a safe natural yellow pigment with a wide range of pharmacological activities,is used both in herbal drugs and as a food coloring agents.Studies have shown that curcumin would suffer from extensive metabolism in vivoand the predominant metabolic pathways are reduction and conjugation.In order to comprehensively study the metabolism and enrich the metabolic profile of cxurcumin in vivo,we carried out this research.A systematic method with highly sensitive UPLC-Q/TOF-MS was established to analyze different biological samples of rats after

Deciphering the chemical profile and pharmacological mechanism of Jinlingzi powder(金铃子散)against bile reflux gastritis using ultra-high performance liquid chromatography coupled with Q exactive focus mass spectrometry,network pharmacology,and molecular docking

OBJECTIVE:To elucidate the chemical profile and the pharmacological mechanism by which Jinlingzi powder(金铃子散,JLZP)treats bile reflux gastritis(BRG).METHODS:A BRG model was established in rats by oral administration of the model solution.JLZP was orally administered for 35 d.Residual gastric rate and tumor necrosis factor(TNF)-α,interleukin(IL)-6,and gastrin levels in the serum were measured,and stomach tissues were collected for histopathological analysis.We used ultra-high performance liquid chromatography coupled with Q Exactive Focus mass spectrometry to identify the chemical ingredients in JLZP.Then,protein-protein interaction and herb-compound-target networks were constructed to screen potential bioactive compounds and targets.Kyoto Encyclopedia of Genes and Genomes pathway analysis was then performed to elucidate the pathway involved in the JLZP-mediated treatment of BRG.After constructing the core compound-target-pathway interaction network,molecular docking was performed to study the binding free energy of core bioactive compounds and two candidate targets[RAC-alpha serine/threonine-protein kinase(AKT1)and phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform(PIK3CA)].RESULTS:JLZP extracts significantly promoted gastric emptying,regulating the release of cytokines(TNF-αand IL-6)and improving gastrin secretion and mucosal repair.Fifty-six compounds were tentatively characterized in JLZP.Moreover,the network pharmacology and molecular docking results showed that alkaloids and flavonoids might be the bioactive compounds in JLZP that treat BRG.JLZP might improve mucosal repair during BRG progression by modulating the phosphatidylinositol-4,5-bisphosphate 3-kinase-protein kinase B,hypoxia inducible factor-1,mitogen-activated protein kinase,forkhead box O,TNF,and IL-17 signaling pathways.CONCLUSIONS:We elucidated the chemical constituents and the pharmacological mechanism of JLZP in treating BRG and provided a basis for clinical application.

Urinary metabolomics analysis of the anti-depressive effects of Hemerocallis citrina extracts in a simulated microgravity-induced rat model of depression

We investigated the antidepressant-like activity of Hemerocallis citrine Baroni extract(HCE)in a simulated microgravity(SMG)-induced rat model of depression using a metabolomics method.A rat model,generated via 14 d of SMG induction,was validated from the reduced sucrose preference and the enhanced immobility time in the forced swimming test.HCE and paroxetine reversed certain metabolite profiles.Anti-depressant effects of HCE might involve the regulation of several metabolic pathways,such as phenylalanine,glutamic acid,and tryptophan metabolism and changes in energy metabolism.5-Hydroxytryptophan,hippuric acid,phenylacetylglycine,citric acid,3-hydroxykynurenine,cyclic AMP,and L-DOPA profiles were altered upon HCE and paroxetine administration.Furthermore,glutamic acid was only regulated in the HCE group,while xanthurenic acid and deoxyuridine were reversed in the positive group,suggesting differences in the mechanisms between the positive drugs and HCE in improving glutamic acid metabolism.This study provided a theoretical foundation for the application of HCE in depression therapy.

Screening Safflower Injection for Constituents with Activity against Stroke Using Comprehensive Chemical Profiling Coupled with Network Pharmacology

Objective:This study aimed to explore safflower injection(SI)for constituents with activity against ischemic stroke using a combination of chemical analysis and a network pharmacology strategy.Materials and Methods:The main ingredients of SI were comprehensively identified using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry,and the core targets and pathways associated with stroke were predicted using PharmMapper and Kyoto Encyclopedia of Genes and Genomes analysis.Cytoscape software was used to visualize and analyze the active compound-target-pathway network of SI regulating ischemic stroke.Results:A total of76 chemical compounds were identified from the SI sample,including 63,which regulated 88 targets that were ultimately enriched in 12 key ischemia stroke-related signaling pathways.Kaempferol-3-O-sophoroside,kaempferol-3-O-rutinoside,carthamoside B6,neoeriocitrin,and6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside were determined to be important for stroke treatment because they had a higher degree value in the network than other constituents did.Moreover,the characteristic components isolated from SI showed protective effect mainly by acting on multiple targets including AKT1,epidermal growth factor receptor,transforming growth factor-beta receptor(TGFBR),Ras homolog,mTORC1 binding,caspase 3,and glycogen synthase kinase 3 beta,which were involved in different signaling pathways including phosphoinositide 3-kinase-Akt,mitogen-activated protein kinase,neurotrophin,ErbB,mechanistic target of rapamycin,and tumor necrosis factor.Conclusions:This study proposed a network pharmacology and chemical component profiling strategy for the systematic understanding of the therapeutic material basis of using SI against ischemic stroke.

Analysis and Identification of Chemical Constituents of Fenugreek by UPLC-IT-MSn and UPLC-Q-TOF-MS

Fenugreek, a traditional Chinese medical plant, has been widely used as food ingredients because of its out- standing medicinal qualities. The hypoglycemic activity of fenugreek is one of its important pharmacological properties. Both of saponin and flavonoid components have the hypoglycemic activity and their contents in fenugreek are 4%--8% and 1% ---2%, respectively. This paper focused on these two types of components to carry out purification research and chemical analysis. The heating reflux extraction method and macroporous resin purification method were designed to prepare the saponins and the flavonoids components from defatted fenugreek seeds. Petroleum ether ultrasonic skim and 70% ethanol refluxing were used for the extraction of saponins and flavonoid from fenugreek. The column of DM130 macroporous resin and D101 macroporous resin were respectively used to prepare fenugreek saponin and flavonoid components, and the total contents of them were 78.56% and 62.28%, respectively. The saponin and flavonoids were subsequently analyzed by an ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry（UPLC-Q-TOF-MS） along with an ultra-performance liquid chromatography and ion-trap tandem mass spectrometry（UPLC-IT-MS＂）. As a result, 57 saponins and 19 flavonoid compounds were characterized. The obtained results will provide a theory basis for further research on fenugreek, as well as research and development of hypogly- cemic new drugs.