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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.

Makyo‐kanseki‐to has been used for the treatment of pneumonia, becoming a basic formula for coronavirus disease 2019. However, the chemical profile of Makyo‐kanseki‐to granule and its possible mechanism against acute lung injury from terminal metabolic regulation have been unclear. The aim of this study was to characterize the constituents in Makyo‐kanseki‐to granule and reveal the potential related mechanism of Makyo‐kanseki‐to granule treatment for acute lung injury using a rat model of lipopolysaccharide‐induced acute lung injury. Totally, 78 constituents were characterized based on ultra‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry. Makyo‐kanseki‐to granule could alleviate acute lung injury through modulating rectal temperature, pulmonary edema, histopathology, and processes of inflammatory and oxidative stress. Twenty‐two potential biomarkers in acute lung injury rats were identified by metabolomics based on ultra‐performance liquid chromatography coupled with quadrupole exactive high‐field mass spectrometry. They were mainly involved in amino acids and glycerophospholipid metabolism, which were regulated by Makyo‐kanseki‐to granule. The present results not only increase the understanding of the chemical profile and molecular mechanism of Makyo‐kanseki‐to granule mediated protection against acute lung injury but also provide an experimental basis and new ideas for further development and clinical application of Makyo‐kanseki‐to granule. [ABSTRACT FROM AUTHOR]

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)-α, and prostaglandin E2 (PGE2) 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-α, and PGE2 was 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-α, and PGE2, and the antipyretic mechanism may be related to changes in metabolites and metabolic pathways in the hypothalamus.

The quantification of antibiotics (ABs) in sediments is an analytical challenge, but at the same time, it is indispensable to understand the fate of ABs in aquatic systems such as rivers. The aim of this study was to develop a comprehensive method to determine 19 ABs classified as macrolides, sulfonamides, fluoroquinolones, tetracyclines, clindamycin and trimethoprim in river sediments, using a combination of pressurized liquid extraction and solid phase extraction with the separation and detection with liquid chromatography coupled with mass spectrometry. Our results showed that the physical-chemical properties (e.g., log(Kow) value) of the analytes affected the extraction efficiency. Therefore, we propose to order ABs based on their log(Kow) values instead of traditional classification (macrolides, sulfonamides etc.) to select a suitable extraction solvent. ABs with log(Kow) values below zero (mainly fluoroquinolones and tetracyclines) were difficult to extract with all of the tested protocols compared to ABs with a log(Kow) larger than zero. After comparing different extraction protocols for ABs from solid and sediments, we concluded that recoveries in the range of 0.8 to 64.8% could be achieved for ABs with a log(Kow) value larger than zero using a mixture of acetonitrile and 50 mM phosphoric acid (50/50, v/v) in two extraction cycles at 100 °C. [ABSTRACT FROM AUTHOR]

Interstitial cystitis (IC) is a clinical syndrome characterized by frequency, urgency, and bladder pain or pelvic pain; however, the underlying pathophysiological mechanisms and diagnostic markers are unknown. In this study, microbiome and metabolome analysis were used to explain the urine signatures of IC patients. Urine samples from 20 IC patients and 22 control groups were analyzed by using 16S rRNA sequence and liquid chromatography coupled with mass spectrometry. Four opportunistic pathogen genera, including Serratia, Brevibacterium, Porphyromonas, and Citrobacter, were significantly upregulated in IC group. The altered metabolite signatures of the metabolome may be related to sphingosine metabolism, amino acid metabolism, and fatty acid biosynthesis. Meanwhile, the associations were observed between different metabolites and microbiomes of IC. The present study suggests that the combined signatures of IC in urine microbiome and metabolome may become its prospective diagnostic markers. [ABSTRACT FROM AUTHOR]

Interstitial cystitis (IC) is a clinical syndrome characterized by frequency, urgency, and bladder pain or pelvic pain; however, the underlying pathophysiological mechanisms and diagnostic markers are unknown. In this study, microbiome and metabolome analysis were used to explain the urine signatures of IC patients. Urine samples from 20 IC patients and 22 control groups were analyzed by using 16S rRNA sequence and liquid chromatography coupled with mass spectrometry. Four opportunistic pathogen genera, including Serratia, Brevibacterium, Porphyromonas, and Citrobacter, were significantly upregulated in IC group. The altered metabolite signatures of the metabolome may be related to sphingosine metabolism, amino acid metabolism, and fatty acid biosynthesis. Meanwhile, the associations were observed between different metabolites and microbiomes of IC. The present study suggests that the combined signatures of IC in urine microbiome and metabolome may become its prospective diagnostic markers.

The present investigation showed that each of the three different liquid chromatography modes may be successfully used for the qualitative analysis of nusinersen metabolites in a patient's serum sample extract. However, the smallest number was detected by the hydrophilic interaction liquid chromatography. Furthermore, the response of the mass spectrometry is several times greater for ion pair chromatography compared to reversed-phase one. Various extraction methods were applied for the extraction of nusinersen metabolites from serum. Silica with bonded capture strand for hybridization was applied, as well as silica modified with amino and carboxyl groups for dispersive solid phase extraction. The hybridization allows selective extraction of nusinersen analogs, however, it fails in extraction of short metabolites. On the contrary, the efficiency of weak ion exchange-based extraction was high, even in the case of the direct extraction of nusinersen metabolites from diluted serum samples without a protein removal step. The new material is a great alternative to liquid-liquid extraction and hybridization for the isolation of nusinersen metabolites from the serum of patients with spinal muscular atrophy (SMA). It is a very simple method that uses a low concentration of organic salt and desorption occurs after changing its pH. Such complex studies were performed for the first time for nusinersen metabolites extracted from the serum of SMA patients treated with Spinraza. [Display omitted] • IP RP HPLC gave higher sensitivity and number of metabolites compared to RP HPLC, HILIC • Hybridization failed for extraction of very short nusinersen metabolites from serum • Successful extraction of nusinersen metabolites directly from serum for new material • New weak anion-exchange material is agreat alternative to LLE and hybridization • The newly developed oligo extraction from serum procedure is environmentally friendly [ABSTRACT FROM AUTHOR]

Objective: To elucidate the chemical profile and the pharmacological mechanism by which Jinlingzi powder (, 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-α and 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.

Abstract: Bavachinin (BVC), one of the main bioactive prenylated flavonoids derived from Psoralea corylifolia Linn, has a wide variety of pharmacological effects, such as antiangiogenic, antitumor, antiallergic, anti‐inflammatory and antibacterial activities, especially as a pan‐peroxisome proliferator‐activated receptors agonist. A rapid and sensitive method for quantifying BVC in rat plasma was developed and validated through ultra‐high‐performance liquid chromatography coupled with electrospray‐ionization tandem mass spectrometry. Furthermore, a complete metabolic investigation of BVC was performed through ultra‐high‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry. In the pharmacokinetic analysis, BVC exhibited rapid oral absorption (Tmax = 0.68 ± 0.21 h), good elimination (T1/2 = 2.27 ± 1.63 h) following oral administration and poor absolute bioavailability (5.27%). Moreover, 11 metabolites of BVC in plasma, urine, bile and feces were characterized. The main metabolic pathways of BVC involved isomeriszation, glucuronidation, sulfonation, hydroxylation, methoxylation and reduction. In conclusion, the present study provides a sensitive quantitative method with a lower limit of quantification of 1 ng/mL and an improved comprehension of the physiological disposition of BVC. [ABSTRACT FROM AUTHOR]

Summary: Brown seaweeds contain a number of bioactive compounds. The xanthophyll, fucoxanthin, has in vivo efficacy against disorders such as type 2 diabetes, obesity and cancer. Organic solvents are traditionally employed to extract fucoxanthin, but carry a toxic chemical and environmental burden. The aim of this study was to optimise a fucoxanthin extraction method using enzymes, water, low‐temperature dehydration and mechanical blending, to produce yields comparable to those achieved with an organic solvent (acetone). Response surface methodology was applied, using Fucus vesiculosus as a model species. A fucoxanthin yield of 0.657 mg g−1 (dry mass) was obtained from F. vesiculosus blade using the enzymatic method, equivalent to 94% of the acetone‐extracted yield. Optimum extraction parameters were determined to be enzyme‐to‐water ratio 0.52%, seaweed‐to‐water ratio 5.37% and enzyme incubation time 3.05 h. These findings may be applied to the development of value‐added nutraceutical products from seaweed. [ABSTRACT FROM AUTHOR]

Licorice ( Glycyrrhiza uralensis Fisch.) is one of the most popular herbal medicines worldwide. This study aims to identify the metabolites of seven representative bioactive licorice compounds in rats. These compounds include 22β-acetoxyl glycyrrhizin ( 1 ), licoflavonol ( 2 ), licoricidin ( 3 ), licoisoflavanone ( 4 ), isoglycycoumarin ( 5 ), semilicoisoflavone B ( 6 ), and 3-methoxy-9-hydroxy-pterocarpan ( 7 ). After oral administration of 250 mg/kg of 1 or 40 mg/kg of 2 – 7 to rats, a total of 16, 43 and 31 metabolites were detected in the plasma, urine and fecal samples, respectively. The metabolites were characterized by HPLC/DAD/ESI–MS n and LC/IT-TOF-MS analyses. Particularly, two metabolites of 1 were unambiguously identified by comparing with reference standards, and 22β-acetoxyl glycyrrhizin-6″-methyl ester ( 1 - M2 ) is a new compound. Compound 1 could be readily hydrolyzed to eliminate the glucuronic acid residue. The phenolic compounds ( 4 – 7 ) mainly undertook phase II metabolism (glucuronidation or sulfation). Most phenolic compounds with an isoprenyl group (chain or cyclized, 2–5 ) could also undertake hydroxylation reaction. This is the first study on in vivo metabolism of these licorice compounds. [ABSTRACT FROM AUTHOR]

Zhi-Zi-Da-Huang decoction (ZZDHD), a classical traditional Chinese medicine (TCM) prescription composed of four herbal medicines, has been widely used in treating various hepatobiliary disorders for a long time. The objective of this study was to develop a sensitive and efficient liquid chromatography coupled with mass spectrometry (LC-MS) method for quantitative determination of 20 active constituents, including three iridoid glycosides, 11 flavonoids, three anthraquinones and three tannins in ZZDHD. Separation was achieved on a phenomenex kinetex C18 column (150 × 4.6 mm, 2.6 µm) using gradient elution with a mobile phase consisting of acetonitrile and 0.1% formic acid in water. Detection was performed with electrospray ionization source in the negative ionization and selected ion monitoring mode. The established method was validated by determining the linearity ( r2 ≥ 0.9983), limit of quantification (0.16-300 ng/mL), precision (RSD ≤ 4.6%), average recovery (96.0-105.6%), repeatability (RSD ≤ 3.2%) and stability (RSD ≤ 4.5%). Then, the method was successfully applied to investigate the chemical composition variations owing to the interaction between the four component herbs of ZZDHD during the extraction process. It was found that different combinations of the herbs affect the extraction efficiency of chemical constituents in different ways. The validated LC-MS method provides a meaningful basis for quality control and further research on ZZDHD. Copyright © 2015 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

To discover new natural compounds from herbal medicines tends to be more and more difficult. In this paper, a strategy integrating orthogonal column chromatography and liquid chromatography/mass spectrometry (LC/MS) analysis was proposed, and was applied for rapid discovery of new ginsenosides from Panax ginseng (PG), Panax quinquefolium (PQ), and Panax notoginseng (PN). The ginsenosides extracts were fractionated by MCI gel×silica gel orthogonal column chromatography. The fractions were then separated on a C18 HPLC column, eluted with a three-component mobile phase (CH3CN/CH3OH/3 mM CH3COONH4 = H2O), and detected by electrospray ionization tandem mass spectrometry. The structures of unknown ginsenosides were elucidated by analyzing negative and positive ion mass spectra, which provided complementary information on the sapogenins and oligosaccharide chains, respectively. A total of 623 comprising 437 potential new ginsenosides were characterized from the ethanol extracts of PG, PQ and PN. New acylations, diversified saccharide chains and C-17 side chains constituted novelty of the newly identified ginsenosides. An interpretation guideline was proposed for structural characterization of unknown ginsenosides by LC/MS. To confirm reliability of this strategy, two targeted unknown trace ginsenosides were obtained in pure form by LC/MS-guided isolation. Based on extensive NMR spectroscopic analysis and other techniques, they were identified as 3-O-[6-O-(E)-butenoyl-β-D-glucopyranosyl(1,2)-β-D-glucopyranosyl]-20(S)-protopanaxadiol-20-O-β-D-glucopyranosyl(1,6)-β-D-glucopyranoside (named ginsenoside IV) and 3-O-β-D-glucopyranosyl(1,2)-β-D-glucopyranosyl-3β,12β,20(S),24(R)-tetra hydroxy-dammar-25-ene-20-O-β-D-glucopyranosyl(1,6)-β-D-glucopyranoside (ginsenoside V), respectively. The fully established structures were consistent with the MS-oriented structural elucidation. This study expanded our understanding on ginsenosides of Panax species, and the proposed strategy was proved efficient and reliable in the discovery of new minor compounds from herbal extracts. [ABSTRACT FROM AUTHOR]

Abstract: Entropy-based methods have been extensively used to measure the uncertainty information in a variety of fields. In this article, a novel information theory-based method for reducing noise of liquid chromatography–mass spectrometry (LC/MS) data was developed. The uncertainty existed in the LC/MS chromatograms was captured and evaluated by information entropy. By comparing the information entropy computationally derived from mass chromatograms, the good quality chromatograms and the noisy chromatograms can be distinguished. The proposed method was applied in processing LC/MS data of “Jing–Zhi–Guan–Xin” troche which is a well-known preparation of traditional Chinese medicine (TCM). The obtained result indicated that this method is beneficial to reduce noise of LC/MS data of complicated chemical samples, such as TCM. [Copyright &y& Elsevier]

As a traditional Chinese medicine (TCM) formula, Wu-tou decoction has been used for treating rheumatoid arthritis (RA) for more than a thousand years. Identifying pharmacodynamic constituents (PCs) of WTD and exploring their in vivo process are very meaningful for promoting the modernization of TCM. However, the pathological state might change this process. Hence, it is necessary and significant to compare the process in vivo of drugs both in normal and disease state and clarify their action mechanism. Taking Wu-tou decoction (WTD) as the research object, a comprehensive strategy based on liquid chromatography coupled with mass spectrometry (LC-MS) was developed to identify PCs, clarify and compare their absorption and distribution in normal and model rats, and then explore the potential mechanism of TCM. Firstly, the PCs in WTD were identified. Then, the pharmacokinetics (PK) and tissue distribution of these ingredients were studied. Finally, the constituents with the difference between normal and model rats were selected for target network pharmacological analysis to clarify the mechanism. A total of 27 PCs of WTD were identified. The absorption and distribution of 20 PCs were successfully analyzed. In the disease state, the absorption and distribution of all these components were improved to have better treatment effects. The results of target network pharmacological analysis indicated that PTGS1, PTGS2, ABCB1, SLC6A4, CHRM2, ESR1, ESR2, CDK2, TNF and IL-6 are 10 key targets for WTD against RA. The regulatory effects of WTD on the expression of PTGS2 and TNF were further verified. Pathway enrichment analysis showed that the key mechanism of WTD against RA is to reduce inflammation and regulate the immune response. These results indicated that this strategy could better understand the in vivo process and mechanism of WTD under the pathological state. Furthermore, this strategy is also appropriate for other TCM. [Display omitted] • The PCs, absorption and distribution of WTD in normal and AIA rats were compared and clarified. • In disease state, the absorption and distribution of PCs were improved to have better treatment effects. • The main mechanism of WTD in treating RA is to regulate inflammation and immune response. [ABSTRACT FROM AUTHOR]

Background: Exploring the pharmacokinetic (PK) changes of various active components of single herbs and their combinations is necessary to elucidate the compatibility mechanism. However, the lack of chemical standards and low concentrations of multiple active ingredients in the biological matrix restrict PK studies., Methods: A putative multiple reaction monitoring strategy based on liquid chromatography coupled with mass spectrometry (LC-MS) was developed to extend the PK scopes of quantification without resorting to the use of chemical standards. First, the compounds studied, including components with available reference standard (ARS) and components lacking reference standard (LRS), were preclassified to several groups according to their chemical structures. Herb decoctions were then subjected to ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry analysis with appropriate collision energy (CE) in MS 2 mode. Finally, multiple reaction monitoring transitions transformed from MS 2 of ultrahigh-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry were used for ultrahigh-performance liquid chromatography coupled with triple quadrupole mass spectrometry to obtain the mass responses of LRS components. LRS components quantification was further performed by developing an assistive group-dependent semiquantitative method., Results: The developed method was exemplified by the comparative PK process of single herbs Radix Ginseng (RG), Radix Polygala (RP), and their combinations (RG-RP). Significant changes in PK parameters were observed before and after combination., Conclusion: Results indicated that Traditional Chinese Medicine combinations can produce synergistic effects and diminish possible toxic effects, thereby reflecting the advantages of compatibility. The proposed strategy can solve the quantitative problem of LRS and extend the scopes of PK studies., (© 2018 The Korean Society of Ginseng. Publishing services by Elsevier B.V.)

Bioactive lipid mediators play crucial roles in promoting the induction and resolution of inflammation. Eicosanoids and other related unsaturated fatty acids have long been known to induce inflammation. These signaling molecules can modulate the circulatory system and stimulate immune cell infiltration into the site of infection. Recently, DHA- and EPA-derived metabolites have been discovered to promote the resolution of inflammation, an active process. Not only do these molecules stop the further infiltration of immune cells, they prompt non-phlogistic phagocytosis of apoptotic neutrophils, stimulating the tissue to return to homeostasis. After the rapid release of lipid precursors from the plasma membrane upon stimulation, families of enzymes in a complex network metabolize them to produce a large array of lipid metabolites. With current advances in mass spectrometry, the entire lipidome can be accurately quantified to assess the immune response upon microbial infection. In this review, we discuss the various lipid metabolism pathways in the context of the immune response to microbial pathogens, as well as their complex network interactions. With the advancement of mass spectrometry, these approaches have also been used to characterize the lipid mediator response of macrophages and neutrophils upon immune stimulation in vitro. Lastly, we describe the recent efforts to apply systems biology approaches to dissect the role of lipid mediators during bacterial and viral infections in vivo., (Copyright © 2013 Elsevier Ltd. All rights reserved.)