ABSTRACT
ObjectiveTo establish a qualitative analysis method for the chemical constituents of the reference sample of Xiao Xumingtang, and to establish the fingerprint of 15 batches of Xiao Xumingtang, so as to evaluate the quality consistency among batches. MethodAccording to the key information of Xiao Xumingtang in the Key Information Table of Ancient Famous Classical Formulas(25 Formulas), the reference sample of this formula was prepared, and it was detected by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS). The chemical components were identified by self-constructed database, consulting relevant literature, and comparing with the reference substances, and the components were assigned by comparing with single drug samples and negative samples lacking single drug. The fingerprint of the reference sample of Xiao Xumingtang was established using high performance liquid chromatography(HPLC), and the common peaks were assigned and identified through single drug samples and negative samples lacking single drug. ResultBased on the information of MS fragments, relevant literature, and database retrieval, a total of 64 compounds were identified and inferred from the reference sample of Xiao Xumingtang, including 31 flavonoids, 8 terpenoids, 12 triterpenoid saponins, 2 phthalides, 3 phenylpropanoids, 2 gingerols, 5 alkaloids, and 1 cyanoside. Among them, 21 were derived from Scutellariae Radix, 10 from stir-fried Glycyrrhizae Radix et Rhizoma, 9 from Ginseng Radix et Rhizoma, 8 from Paeoniae Radix Alba, 4 from Saposhnikoviae Radix, 3 from Stephaniae Tetrandrae Radix, 3 from Chuanxiong Rhizoma, 2 from Aconiti Lateralis Radix Praeparata, 2 from Zingiberis Rhizoma Recens, 1 from Ephedrae Herba, and 1 from Armeniacae Semen Amarum. The established HPLC fingerprint of the reference sample of Xiao Xumingtang had 23 common peaks, among which, peaks 1 and 2 were derived from Paeoniae Radix Alba, peaks 3 and 7 from Saposhnikoviae Radix, peaks 4, 8 and 9 from Stephaniae Tetrandrae Radix, peaks 10, 17, 18, 20 and 21 from stir-fried Glycyrrhizae Radix et Rhizoma, peaks 11-16, 19 and 22 from Scutellariae Radix, peak 5 from Chuanxiong Rhizoma, peak 23 from Zingiberis Rhizoma Recens, peak 6 was the common component of stir-fried Glycyrrhizae Radix et Rhizoma and Scutellariae Radix. A total of 10 compounds including albiflorin(peak 1), paeoniflorin(peak 2), cimicifugoside(peak 3), 5-O-methylvisammioside(peak 7), baicalin(peak 11), sec-O-glucosylhamaudol(peak 13), oroxylin A-7-O-β-D-glucuronide(peak 15), wogonoside(peak 16), glycyrrhizic acid(peak 21) and 6-gingerol(peak 23) were identified. The similarities of 15 batches of reference samples were>0.999, indicating that the reference samples had good consistency. ConclusionThrough the identification of the chemical constituents in the reference sample of Xiao Xumingtang, it is clear that the composition of the samples is mainly composed of flavonoids and triterpenoid saponins. The established fingerprint can basically reflect the overall chemical characteristics of the reference sample of Xiao Xumingtang, which can provide a basis for the quality research of its compound preparations.
ABSTRACT
ObjectiveTo establish a qualitative and quantitative analysis method for chemical constituents in Liu Junzitang(LJZT), and to clarify its material basis. MethodThe chemical constituents in LJZT were analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS), and the resulting compounds were identified by using databases, such as MassBank, PubChem, ChemSpider, Traditional Chinese Medicine Systems Pharmacology Database and Analytical Platform(TCMSP), and by combining with relevant literature. UPLC was used to establish a quantitative method for analysis of 9 compounds in LJZT, including liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ. ResultBy combining the relevant literature, database and MS information, a total of 79 compounds were identified from LJZT, including 31 flavonoids, 15 terpenoids, 14 nitrogen-containing compounds, 6 phenylpropanoids, 6 organic acids and 7 other compounds. The established quantitative analytical method for the nine representative components showed good linearity within their respective linear ranges, and the precision, stability, reproducibility and recovery were in accordance with the requirements. The quantitative results showed that the contents of liquiritin, hesperidin, lobetyolin, liquiritigenin, glycyrrhizic acid, nobiletin, tangeretin, atractylenolide Ⅱ and Ⅰ in LJZT were 0.376 5, 2.602 1, 0.082 6, 0.128 1, 1.778 6, 0.015 7, 0.006 7, 0.030 4, 0.003 2 mg·g-1, respectively. ConclusionThe established method can quickly, sensitively and accurately analyze the chemical constituents in LJZT, clarify that the material basis of LJZT is mainly flavonoids, terpenoids and nitrogen-containing compounds, and simultaneously determine the contents of the 9 components, which can lay a foundation for the research on quality control, mechanism and clinical application of LJZT.
ABSTRACT
This study aimed to explore the mechanism of Qilongtian Capsules in treating acute lung injury(ALI) based on network pharmacology prediction and in vitro experimental validation. Firstly, UPLC-Q-TOF-MS/MS was used to analyze the main chemical components of Qilongtian Capsules, and related databases were used to obtain its action targets and ALI disease targets. STRING database was used to build a protein-protein interaction(PPI) network. Metascape database was used to conduct enrichment analysis of Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG). AutoDock software was used to perform molecular docking verification on the main active components and key targets. Then, the RAW264.7 cells were stimulated with lipopolysaccharide(LPS) for in vitro experiments. Cell viability was measured by MTT and ROS level was measured by DCFH-DA. NO content was measured by Griess assay, and IL-1β, IL-6, and TNF-α mRNA expression was detected by RT-PCR. The predicted targets were preliminarily verified by investigating the effect of Qilongtian Capsules on downstream cytokines. Eighty-four compounds were identified by UPLC-Q-TOF-MS/MS. Through database retrieval, 44 active components with 589 target genes were screened out. There were 560 ALI disease targets, and 65 intersection targets. PPI network topology analysis revealed 10 core targets related to ALI, including STAT3, JUN, VEGFA, CASP3, and MMP9. KEGG enrichment analysis showed that Qilongtian Capsules mainly exerted an anti-ALI effect by regulating cancer pathway, AGE-RAGE, MAPK, and JAK-STAT signaling pathways. The results of molecular docking showed that the main active components in Qilongtian Capsules, including crenulatin, ginsenoside F_1, ginsenoside Rb_1, ginsenoside Rd, ginsenoside Rg_1, ginsenoside Rg_3, notoginsenoside Fe, notoginsenoside G, notoginsenoside R_1, notoginsenoside R_2, and notoginsenoside R_3, had good binding affinities with the corresponding protein targets STAT3, JUN, VEGFA, CASP3, and MMP9. Cellular experiments showed that Qilongtian Capsules at 0.1, 0.25, and 0.5 mg·mL~(-1) reduced the release of NO, while Qilongtian Capsules at 0.25 and 0.5 mg·mL~(-1) reduced ROS production, down-regulated mRNA expression of IL-1β, IL-6, TNF-α, and inhibited the inflammatory cascade. In summary, Qilongtian Capsules may exert therapeutic effects on ALI through multiple components and targets.
Subject(s)
Humans , Tumor Necrosis Factor-alpha , Ginsenosides , Caspase 3 , Matrix Metalloproteinase 9 , Interleukin-6 , Molecular Docking Simulation , Network Pharmacology , Reactive Oxygen Species , Tandem Mass Spectrometry , Acute Lung Injury/genetics , Capsules , RNA, Messenger , Drugs, Chinese Herbal/pharmacologyABSTRACT
The Compound Cheqian Tablets are derived from Cheqian Power in Comprehensive Recording of Divine Assistance, and they are made by modern technology with the combination of Plantago asiatica and Coptis chinensis. To investigate the material basis of Compound Cheqian Tablets in the treatment of diabetic nephropathy, in this study, the chemical components of Compound Cheqian Tablets were characterized and analyzed by UPLC-Q-TOF-MS/MS, and a total of 48 chemical components were identified. The identified chemical compounds were analyzed by network pharmacology. By validating with previous literature, six bioactive compounds including acteoside, isoacteoside, coptisine, magnoflorine, palmatine, and berberine were confirmed as the index components for qua-lity evaluation. Furthermore, the content of the six components in the Compound Cheqian Tablets was determined by the "double external standards" quantitative analysis of multi-components by single marker(QAMS), and the relative correction factor of isoacteoside was calculated as 1.118 by using acteoside as the control; the relative correction factors of magnoflorine, palmatine, and berberine were calculated as 0.729, 1.065, and 1.126, respectively, by using coptisine as the control, indicating that the established method had excellent stability under different conditions. The results obtained by the "double external standards" QAMS approximated those obtained by the external standard method. This study qualitatively characterized the chemical components in the Compound Cheqian Tablets by applying UPLC-Q-TOF-MS/MS and screened the pharmacodynamic substance basis for the treatment of diabetic nephropathy via network pharmacology, and primary pharmacodynamic substance groups were quantitatively analyzed by the "double external stan-dards" QAMS method, which provided a scientific basis for clarifying the pharmacodynamic substance basis and quality control of Compound Cheqian Tablets.
Subject(s)
Humans , Tandem Mass Spectrometry , Berberine/pharmacology , Chromatography, High Pressure Liquid/methods , Network Pharmacology , Diabetic Nephropathies , Drugs, Chinese Herbal/chemistry , Quality Control , TabletsABSTRACT
Aim To explore the mechanism of process¬ing and increasing efficiency of Arisaematis rhizomz preparatum. Methods UPLC-Q-TOF-MS/MS tech¬nology was used to detect the chemical components be¬fore and after processing of Arisaematis rhizomz prepara¬tum, and its mechanism of action was analysed in the treatment of 44 asthma and phlegm " by using network pharmacology. A rat model of allergic asthma was es- tablished to compare the efficacy of Arisaematis rliizoma before and after processing. Results A total of 27 chemical components were identified, among which cur- cumin ,6-gingerol and other components increased after processing. Combined with the database prediction, the action mechanism of the 36 chemical components in the treatment of 44 asthma and phlegm" diseases was dis¬cussed and predicted through network pharmacology. The results of animal experiments showed that the effect of processed Arisaematis rhizoma on allergic asth¬ma was better than that of Arisaematis rhizoma, but there was no significant difference. Conclusions The addition of curcumin, 6-gingerol, camphor, demethyl- curcumin and other components after the processed Ari¬saematis rhizomz preparatum may be the reason for the synergistic effect of Arisaematis rhizomz preparatum in the treatment of allergic asthma.
ABSTRACT
OBJECTIVE To analyze the main components of Chelidonii Herba-Corydalis Rhizoma (CHCR), and to predict pharmacodynamic substances against estrogen receptor (ER) -positive breast cancer and their potential targets and signaling pathways, followed by verifying experiments. METHODS The ethanol extract of CHCR was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS). The network pharmacology analysis was performed for the screened components. The network diagram of CHCR “active components-target-pathway” was constructed, and the enrichment pathway in vitro was validated. RESULTS A total of 58 chemical components were identified, including 57 alkaloids and 1 organic acid. A total of 38 active ingredients were screened from the network pharmacology, and 38 core targets were found in the protein-protein interaction network of “component-disease” intersection targets; 258 gene ontology entries and 137 Kyoto encyclopedia of genes and genomics pathways were obtained, mainly including estrogen signal pathway, phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) signal pathway, etc. The results of validation test showed that the median inhibitory concentration of CHCR to MCF-7 cells was 693 μg/mL; 150, 300, 600 μg/mL CHCR could significantly reduce the expressions of phosphorylated PI3K, phosphorylated Akt, ERα protein and ESR1 mRNA (P<0.01). CONCLUSIONS The anti-ER-positive breast cancer effect of CHCR may be related to the regulation of ER and PI3K/Akt pathways, which has the characteristics of multi-component and multi-target effects.
ABSTRACT
ObjectiveTo analyze the migrating components absorbed into blood of the aqueous extract of Euphorbia helioscopia, and to explore the pharmacodynamic material basis of the aqueous extract of E. helioscopia against chronic obstructive pulmonary disease(COPD). MethodUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) was used to detecte the migrating components absorbed into blood of rats after intragastric administration of aqueous extract of E. helioscopia. An Agilent RRHD SB-C18 column(3 mm×100 mm, 1.8 μm) was used with 0.1% formic acid aqueous solution(A)-acetonitrile(B) as the mobile phase for gradient elution(0-15 min, 5%-30%B; 15-20 min, 30%-50%B; 20-30 min, 50%-95%B; 30-35 min, 95%-5%B), and the detection wavelength of 190-800 nm, column temperature of 40 ℃, flow rate of 0.3 mL∙min-1 and injection volume of 4 μL. The electrospray ionization(ESI) was used in positive and negative ion modes, and the detection range was m/z 50-1 250. Network pharmacology was used to screen out the key components and the key targets of COPD through the interaction analysis. Metascape database was used to predict the molecular function, biological process, cellular composition and signal pathways mainly involved in the anti-COPD effect of E. helioscopia. Molecular docking technique was used to determine the affinity of key targets with key components. ResultA total of 29 migrating components absorbed into blood of rats were identified after intragastric administration of aqueous extract of E. helioscopia, 9 of which were prototype components and 20 were metabolites. Network pharmacological analysis showed that luteolin, quercetin, apigenin, naringenin and helioscopinolide C were the key components of E. helioscopia against COPD, and vascular endothelial growth factor A(VEGFA), albumin(ALB), protein kinase B1(Akt1), tumor necrosis factor(TNF) and interleukin-6(IL-6) were the key targets. Molecular docking results showed that one diterpene lactone(helioscopinolide C) and three flavonoids(naringenin, luteolin, apigenin) in the migrating components absorbed into blood all had strong binding activity to the key targets of E. helioscopia against COPD. ConclusionNaringenin, helioscopinolide C, luteolin and apigenin may be the main anti-COPD active substances of E. helioscopia.
ABSTRACT
In this study, untargeted metabolomics technology based on ultra-high-performance liquid chromatography-quadrupole/time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) was used to analyze and identify the overall chemical components of Juniperri Caulis et Folium. Chemical markers for the identification of different Juniperri Caulis et Folium species were screened by integrated principal component analysis and partial least squares discriminant analysis. A total of 58 chemical components were detected and 46 of them were identified, including 26 flavonoids, 8 organic acids and their derivatives, 4 phenylpropanoids, 3 terpenoids, and 5 other components. Among them, methylsyringin and ekersenin were identified for the first time. In the positive ion mode, 12 markers were screened, and in the negative ion mode, 13 markers were screened for species identification. In summary, UPLC-Q-TOF-MS/MS metabonomics technology combined with chemometrics method can effectively reveal the chemical composition differences of different Juniperri Caulis et Folium species, and provide reference for its species identification and quality control.
ABSTRACT
OBJECTIVE To study the pharmacological basis of Schisandra chinensis in the treatment of allergic asthma. METHODS The common components of 10 batches of S. chinensis from different habitats were analyzed by UPLC-Q-TOF-MS/MS. Furthermore, the allergic asthma model was established by intraperitoneal injection of ovalbumin (OVA) and aluminum hydroxide for stimulation combined with atomization exitation; general behavioral observation and the contents of interferon γ (IFN-γ), interleukin-4 (IL-4) and immunoglobulin E (IgE) in serum were taken as criteria for evaluating the therapeutic effect of S. chinensis from different habitats in the treatment of allergic asthma. Correlation coefficients between common peak area and efficacy evaluation index of each batch of medicinal material were analyzed through grey correlation degree and Pearson correlation analysis. RESULTS A total of 21 common components were identified in 10 batches of S. chinensis from different habitats. After administration of S. chinensis, symptoms such as shortness of breath, sneezing and curling of rats were alleviated. In addition, the content of IFN-γ was significantly increased while the contents of IL-4 and IgE in serum were distinctly decreased (P<0.01). Grey correlation analysis showed that 11 common components had high correlation coefficients with IFN-γ, IL-4 and IgE (rˉ>0.8). Pearson correlation analysis showed that 8 components were significantly positively correlated with the content of IFN-γ (P< 0.05), and 9, 8 components were significantly negatively correlated with the content of IL-4 and IgE (P<0.05). Based on the results of grey correlation degree and Pearson correlation analysis, 7 components such as peak 3, 4, 6, 7, 9, 19 and 20, were highly related to S. chinensis in the treatment of allergic asthma. CONCLUSIONS Schisandrol A, schisandrin B, schisandrin C, gomisin M2, gomisin J, pregomisin and angeloylgomisin H are the potential pharmacodynamic substance basis of S. chinensis in the treatment of allergic asthma.
ABSTRACT
@#Heibu Yaogao (ointment), which made of Galla chinensis, Scolopendra, etc., is a commonly used hospital preparation with definite clinical efficacy, yet its material basis has not been fully clarified.By ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS), the chemical constituents of Heibu Yaogao (ointment) and its crude drug mixture were qualitatively analyzed, and the differences of components before and after preparation were discussed.Scanned by electrospray ionization (ESI) source in positive and negative ion modes, a total of 73 compounds were identified via precise molecular weight, characteristic fragment ions and chromatographic retention time.57 and 65 compounds were detected in crude drug mixture and Heibu Yaogao (ointment) respectively, including tannins, phenolic acids, nucleosides, amino acids, etc.In the process of preparation, the chemical structure changes are mainly concentrated in tannins.The change of components of Heibu Yaogao (ointment) before and after processing were proved by UPLC-Q-TOF-MS/MS, which provides important reference for elucidating the material basis of Heibu Yaogao (ointment), illustrating the rationality of preparation technology and further promoting the quality control of the preparation.
ABSTRACT
ObjectiveTo develop a quality control method for the simultaneous determination of multiple active components in Nymphaeae Flos aiming at the problems of the single index for quality control and the relatively low overall quality control level. MethodUltra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS)was used to identify and select the index components for quality control with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B)for gradient elution (0-2 min, 3%-8%B; 2-4 min, 8%-10%B; 4-13 min, 10%-15%B; 13-19 min, 15%-20%B; 19-26 min, 20%-45%B) at a flow rate of 0.4 mL·min-1, detection wavelength of 350 nm, electrospray ionization(ESI), negative ion scanning mode, ion source temperature of 120 ℃, scanning range of m/z 100-1 200, transmit collision energy of 6 eV for low-energy scanning and 25-50 eV for high-energy scanning. High performance liquid chromatography(HPLC)was used to establish the quality control method for the simultaneous determination of multi-index components with the mobile phase of 0.2% phosphoric acid aqueous solution(A)-acetonitrile(B) for gradient elution(0-30 min, 12%-15%B; 30-60 min, 15%-22%B; 60-90 min, 22%-40%B)and detection wavelength of 350 nm. The preparation method of the test solution for content determination was refluxing extraction for 60 min with 80 times the amount of 70% methanol. ResultBy comparing the retention time, ultraviolet absorption characteristics, MS and MS/MS spectrometric signals in the samples with the reference substances, 8 active components with high contents, including brevifolincarboxylic acid, ellagic acid, rutin, nicotiflorin, astragalin, quercetin, quercetin-3-methylether and kaempferol, were identified qualitatively from Nymphaeae Flos, which were selected as the index components for quality control. Under the established HPLC conditions, the above 8 components could be well separated(resolution>1.5), and showed good linearity(r=0.999 9)between the concentration ranges of 1.99-99.6, 1.76-176, 1.52-75.8, 3.60-180, 0.964-96.4, 1.18-118, 1.94-96.8, 1.04-104 mg·L-1 and the peak areas, respectively. The detection limits of them were 10-49 μg·L-1, and the limits of quantitation were 34-164 μg·L-1. The average recoveries were 97.12%-103.1% with the relative standard deviations (RSDs) were 1.1%-2.2%. ConclusionA quality control method for simultaneous determination of the multiple active components in Nymphaeae Flos have been developed, which is simple, accurate and reproducible, and it can provide a scientific basis for the formulation of quality standard of this herb and lay a research foundation for the transformation of Uygur hospital preparations containing Nymphaeae Flos into new drugs.
ABSTRACT
ObjectiveUsing multi-omics technology, we conducted the present study to determine whether dexamethasone has therapeutic effect on pneumonia rats through the regulation of intestinal flora and metabolites. MethodsTotally 18 Sprague-Dawley rats were randomly divided into 3 groups (n = 6 each): Control group, Model group and Dexamethasone (Dex) group. Lipopolysaccharide (LPS) was continuously injected intraperitoneally into rats at a dose of 4 mg/kg for 7 days to induce pneumonia except the Control group. Then the Dex group was given Dex at a dose of 2 mg/kg via oral gavage for 12 days, and both the other two groups received continuously equal volume of sterile PBS buffer for 12 days. On the 19th day, lung, plasma, feces and intestinal contents of rat were collected. Hematoxylin-eosin (H&E) staining and Bio-plex suspension chip system were applied to evaluate the effect of Dex on pneumonia. Furthermore, metagenomic sequencing and UPLC-Q-TOF-MS/MS technology were employed to determine the intestinal flora and metabolites of rats, respectively. ResultsH&E staining results showed that the lung tissue of the Model group was infiltrated with inflammatory cells, the alveolar septum was increased, alveolar hemorrhage, and histological lesions were less severe in Dex group than in the model group. The levels of 3 inflammatory cytokines including TNF-α (P < 0.000 1), IL-1α (P = 0.009 6) and IL-6 (P < 0.000 1) in the Model group were increased compared with the Control group, while Dex treatment reduced the levels of the three inflammatory factors. Taken together, Dex treatment effectively reversed the features of pneumonia in rats. Metagenomic analysis revealed that the intestinal flora structure of the three groups of rats was changed. In contrast with the Model group, an increasing level of the Firmicutes and an elevated proportion of Firmicutes/Bacteroidetes were observed after Dex treatment. Dex-treated rats possessed notably enrichment of Bifidobacterium, Lachnospiraceae and Lactobacillus. Multivariate statistical analysis showed a great separation between Model group and Dex group, indicating metabolic profile changes. In addition, 69 metabolites (P < 0.05) were screened, including 38 up-regulated in the Model group and 31 elevated in the Dex group, all of which were mainly involved in 3 metabolic pathways: linoleic acid metabolism, tryptophan metabolism and primary bile acid biosynthesis. ConclusionsIn summary, we demonstrate the beneficial effects of Dex on the symptoms of pneumonia. Meanwhile, integrated microbiome-metabolome analysis reveals that Dex improves LPS-induced pneumonia in rats through regulating intestinal flora and host metabolites. This study may provide new insights into the mechanism of Dex treatment of pneumonia in rats.
ABSTRACT
Lilii Bulbus is a commonly used Chinese herbal medicine with both medicinal and edible values, while the market products usually has the problem of sulfur fumigation. Therefore, the quality and safety of Lilii Bulbus products deserve attention. In this study, ultra-high performance liquid chromatography-time of flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was combined with principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA) to analyze the differential components of Lilii Bulbus before and after sulfur fumigation. We identified ten markers generated after sulfur fumigation, summarized their mass fragmentation and transformation patterns, and verified the structures of phenylacrylic acid markers of sulfur fumigation. At the same time, the cytotoxicity of the aqueous extracts of Lilii Bulbus before and after sulfur fumigation was evaluated. The results showed that in the concentration range of 0-800 mg·L~(-1), the aqueous extract of Lilii Bulbus after sulfur fumigation had no significant effect on the viability of human liver LO2 cells, human renal proximal tubular HK-2 cells, and rat adrenal pheochromocytoma PC-12 cells. Moreover, the viability of the cells exposed to the aqueous extract of Lilii Bulbus before and after sulfur fumigation showed no significant difference. This study identified phenylacrylic acid and furostanol saponins as markers of sulfur-fumigated Lilii Bulbus for the first time, and made clear that proper sulfur fumigation of Lilii Bulbus would not produce cytotoxicity, providing a theoretical basis for the rapid identification and quality and safety control of sulfur-fumigated Lilii Bulbus.
Subject(s)
Humans , Animals , Rats , Fumigation , Tandem Mass Spectrometry , Chromatography, High Pressure Liquid , Epithelial Cells , SulfurABSTRACT
The chemical components of Huanglian Decoction were identified by ultra-performance liquid chromatography-quadrupole-time-of-flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS) technology. The gradient elution was conducted in Agilent ZORBAX Extend-C_(18) column(2.1 mm×100 mm, 1.8 μm) with the mobile phase of 0.1% formic acid aqueous solution(A)-acetonitrile(B) at a flow rate of 0.3 mL·min~(-1) and the column temperature of 35 ℃. The MS adopted the positive and negative ion mode of electrospray ionization(ESI), and the MS data were collected under the scanning range of m/z 100-1 500. Through high-resolution MS data analysis, combined with literature comparison and confirmation of reference substances, this paper identified 134 chemical components in Huanglian Decoction, including 12 alkaloids, 23 flavonoids, 22 terpenes and saponins, 12 phenols, 7 coumarins, 12 amino acids, 23 organic acids, and 23 other compounds, and the medicinal sources of the compounds were ascribed. Based on the previous studies, 7 components were selected as the index components. Combined with the network pharmacology research and analysis me-thods, the protein and protein interaction(PPI) network information of the intersection targets was obtained through the STRING 11.0 database, and 20 core targets of efficacy were screened out. In this study, UPLC-Q-TOF-MS/MS technology was successfully used to comprehensively analyze and identify the chemical components of Huanglian Decoction, and the core targets of its efficacy were discussed in combination with network pharmacology, which laid the foundation for clarifying the material basis and quality control of Huanglian Decoction.
Subject(s)
Tandem Mass Spectrometry , Network Pharmacology , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , TechnologyABSTRACT
ObjectiveA rapid method for identification of chemical constituents in Puerariae Lobatae Radix dispensing granules was established in order to clarify the material basis. MethodThe chemical constituents of Puerariae Lobatae Radix dispensing granules was qualitatively analyzed by ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS) under positive and negative ion modes, and the chromatographic conditions were on an ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8 μm) with 0.1% formic acid aqueous solution(A)-0.1% formic acid acetonitrile solution(B) as mobile phase for gradient elution(0-4 min, 5%-10%B; 4-10 min, 10%-15%B; 10-20 min, 15%-16%B; 20-27 min, 16%-31%B; 27-33 min, 31%-59%B; 33-42 min, 59%-95%B; 42-42.1 min, 95%-5%B; 42.1-45 min, 5%B), the flow rate was 0.35 mL·min-1, the column temperature was 40 ℃, the injection volume was 5 μL, and electrospray ionization(ESI) was selected. Then these chemical constituents were comprehensively identified based on PeakView 1.2, PubChem, ChemicalBook, ChemSpider, comparative control profiles and literature information. ResultA total of 128 chemical constituents were identified from the dispensing granules, including 60 flavonoids, 26 organic acids, 7 glycosides, 6 coumarins, 3 nucleosides and 26 other compounds. By focusing on the cleavage patterns of flavonoids, organic acids, glycosides, coumarins, nucleosides and other compounds, 12 compounds that have not been reported in Puerariae Lobatae Radix species were identified from the dispensing granules. ConclusionThe established method can systematically and rapidly identify the chemical constituents in Puerariae Lobatae Radix dispensing granules, and cleared it composition is mainly flavonoids and organic acids. Laying a foundation for the study of the material basis, mechanism of action and clinical application of the dispensing granules.
ABSTRACT
Zhachong Shisanwei Pills, composed of 13 Chinese medicinal materials, are used for treating the diseases such as hemiplegia, pain of muscles and bones, rheumatism, and joint pain. The chemical composition and pharmacodynamics of Zhachong Shisanwei Pills have not been reported. Ultra-performance liquid chromatography/quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to quickly identify the chemical components of Zhachong Shisanwei Pills, which was performed with Shim-pack GIST C_(18) column(4.6 mm×150 mm, 5 μm). The gradient elution was conducted with methanol-0.05% acetic acid as the mobile phase. Electrospray ionization mass spectrometry(ESI-MS) was carried out in both positive and negative ion modes. The compounds were identidied based on accurate relative molecular weight, fragment ion species, and the MS data of reference substances and in literature. In conclusion, a total of 98 compounds were identified, including 19 organic acids, 36 flavonoids, 13 volatile oils, 8 tannins, 5 2-(2-phenylethyl)chromones, 5 amino acids, 3 sesquiterpenoids, 3 alkaloids, and 2 other compounds. This study characte-rized the chemical components of Zhachong Shisanwei Pills rapidly for the first time, laying a foundation for further research on the pharmacodynamic material basis and quality evaluation.
Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Liquid , Drugs, Chinese Herbal/chemistry , Spectrometry, Mass, Electrospray Ionization , Tandem Mass SpectrometryABSTRACT
Suanzaoren Decoction(SZRD) is a classical formula for the clinical treatment of insomnia. This study analyzed the effect of SZRD on endogenous metabolites in insomnia rats based on metabonomics and thereby explored the anti-insomnia mechanism of SZRD. To be specific, DL-4-chlorophenylalanine(PCPA) was used to induce insomnia in rats. Then pathological changes of the liver and brain were observed and biochemical indexes such as 5-hydroxytryptamine(5-HT), dopamine(DA), glutamate(Glu), γ-aminobutyric acid(GABA), and norepinephrine(NE) in the hippocampus and prostaglandin D2(PGD2), tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and IL-6 in the serum of rats were detected. On this basis, the effect of SZRD on PCPA-induced insomnia rats was preliminarily assessed. The metabolic profile of rat serum samples was further analyzed by ultra-performance liquid chromatography-quadrupole-time of flight-tandem mass spectrometry(UPLC-Q-TOF-MS/MS). Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were combined with t-test and variable importance in projection(VIP) to identify differential metabolites, and MetaboAnalyst 5.0 was employed for pathway analysis. The results showed that SZRD could improve the pathological changes of brain and liver tissues, increase the levels of neurotransmitters 5-HT, DA, and GABA in hippocampus and the level of PGD2 in hypothalamic-pituitary-adrenal axis(HPA axis), and reduce the levels of IL-1β and TNF-α in serum of insomnia rats. Metabonomics analysis yielded 12 significantly changed potential metabolites: 5-aminovaleric acid, N-acetylvaline, L-proline, L-glutamate, L-valine, DL-norvaline, D(-)-arginine, pyroglutamic acid, 1-methylguanine, L-isoleucine, 7-ethoxy-4-methylcoumarin, and phthalic acid mono-2-ethylhexyl ester(MEHP), which were related with multiple biochemical processes including metabolism of D-glutamine and D-glutamate, metabolism of alanine, aspartate, and glutamate, metabolism of arginine and proline, arginine biosynthesis, glutathione metabolism. These metabolic changes indicated that SZRD can improve the metabolism in insomnia rats by regulating amino acid metabolism.
Subject(s)
Animals , Rats , Chromatography, High Pressure Liquid/methods , Drugs, Chinese Herbal , Hypothalamo-Hypophyseal System , Metabolomics/methods , Pituitary-Adrenal System , Sleep Initiation and Maintenance Disorders/drug therapy , Tandem Mass SpectrometryABSTRACT
With the ultra high performance liquid chromatography-quadruple-electrostatic field orbitrap high resolution mass spectrometry(UHPLC-Q Exactive Orbitrap-MS)-based metabonomics technology, this study aims to analyze the effect of Chaiqin Ningshen Granules(CNG) on endogenous metabolites in insomnia rats of liver depression syndrome and explore the sleep-improving mechanism of this prescription. Parachlorophenylalanine(PCPA, ip) and chronic stimulation were combined to induce insomnia of liver depression pattern in rats, and the effect of CNG on the macroscopic signs, hemorheology, and neurotransmitters in the hippocampus of insomnia rats of liver depression syndrome was observed. After the administration, rat hippocampus was collected for liquid chromatography-mass spectrometry(LC-MS) analysis of the metabolomics. Principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA), and orthogonal partial least squares discriminant analysis(OPLS-DA) were employed for analyzing the metabolites in rat hippocampus and screening potential biomarkers. MetPA was used to yield the related metabolic pathways and metabolic networks. The results show that the drugs can significantly improve the mental state, liver depression, and blood stasis of rats, significantly increase the content of 5-hydroxytryptamine(5-HT) and gamma aminobutyric acid(GABA) in hippocampus(except low-dose CNG), and significantly reduce the content of glucose(Glu)(except low-dose CNG). Among them, estazolam and high-dose CNG had better effect than others. Metabolomics analysis yielded 27 potential biomarkers related to insomnia. MetPA analysis showed 4 metabolic pathways of estazolam in intervening insomnia and 3 metabolic pathways of high-dose CNG in intervening insomnia, involving purine metabolism, glycerophospholipid metabolism, histidine metabolism, and caffeine metabolism. CNG can alleviate insomnia by regulating endogenous differential metabolites and further related metabolic pathways. The result lays a basis for further elucidating the mechanism of CNG in improving sleep.
Subject(s)
Animals , Rats , Biomarkers , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/pharmacology , Estazolam , Hippocampus/metabolism , Metabolomics/methods , Sleep , Sleep Initiation and Maintenance Disorders/drug therapyABSTRACT
OBJECTIVE To study the absorbed components of Xiebai powder in blood. METHODS UPLC-Q-TOF-MS/MS method was adopted. SD rats were randomly divided into blank group and administration group ,with 10 rats in each group. Blank group was given water intragastrically ,and administration groups were given 2 g/mL(by the amount of crude drug )Xiebai powder solution intragastrically. Administration volume was 11.3 mL/kg,twice a day for 3 days. One point five hours after last administration,blood was taken from the abdominal aorta of each rat ,the serum was processed to obtain the supernatant for analysis;the relevant data in positive and negative ion mode were collected ,and the absorbed components of Xiebai powder in blood were analyzed and identified by using self-built secondary mass spectrometry database and consulting the relevant literature. RESULTS Totally 17 components from Xiebai powder were identified ,among which 6 components came from sovereign Moru salba,7 from minister Cortex Lycii ,12 from assistant Glycyrrhiza uralensis ,i.e. kukoamine A ,chlorogenic acid ,tachiogroside B,astringin,neoglycyrrhizin,glycyrrhizin,azelaic acid ,isoglycyrrhizin,glycyroside,anthocyanin,sebacic acid ,parthenolide, anthocyanin,18β-glycyrrhetinic acid ,6-gingerol,palmitoamide,erucamide. These compounds were mainly flavonoids ,alkaloids and organic acids. CONCLUSIONS In this study ,17 absorbed components of Xiebai powder in blood are preliminarily determined,which are consistent with the effect of Xiebai powder. They may be the pharmacodynamic substances of Xiebai powder.
ABSTRACT
@#Fufang Shechuangzi Xiji (Lotion) is a common hospital preparation made from clinical prescriptions. Its clinical efficacy is accurate, yet its pharmacodynamic material basis has not been clarified.An ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) technique was developed to identify the chemical constituents of Fufang Shechuangzi Xiji (Lotion).The chemical constituents of Fufang Shechuangzi Xiji (Lotion) were scanned by electrospray ionization (ESI) source in positive and negative ion modes.As a result, a total of 118 compounds were identified and characterized via reference standards and by comparing mass spectrometry data with literature, including 45 alkaloids, 21 coumarins and chromones, 19 flavonoids, 14 saponins, 10 anthraquinones and 9 organic acids.As a result, UPLC-QTOF-MS/MS technology can quickly and sensitively identify the chemical constituents of Fufang Shechuangzi Xiji (Lotion), which provides a useful reference for exploring the pharmacodynamic material basis and further quality control study of the preparation.