Chemical Metabolites Research Articles

The integrated study on the chemical profiling and in vivo course to explore the bioactive constituents and potential targets of Chinese classical formula Qingxin Lianzi Yin Decoction by UHPLC-MS and network pharmacology approaches

Ethnopharmacological relevanceQingxin Lianzi Yin Decoction (QXLZY), a Chinese classical formula, has been widely used in the treatment of various chronic kidney diseases over 1,000 years. However, the current studies on QXLZY were mostly focused on its clinical efficacy, lacking systematic material basis research on constituents. Aim of the studyThis work aims to elucidate and quantify the chemical constituents, clarify the blood-absorbed components and excretion pathways, predict major bioactive constituents and discover potential therapeutic targets. Materials and methodsUHPLC-LTQ-Orbitrap HRMS was employed to clarify the chemical constituents and metabolites of QXLZY. The extraction of diagnostic ion and neutral loss fragment was aimed for searching specific type of constituents. The plasma, urine, bile and feces samples of rats after oral administration of QXLZY were systematically studied. UHPLC-QQQ-MS/MS was employed to simultaneously detect different types of constitutes. Based on the analysis of ingredients in vivo, the bioactive constituents and potential therapeutic targets in the treatment of diabetic nephropathy (DN) was investigated by using network pharmacological analysis. ResultsTotally, 220 compounds were identified or tentatively characterized by UHPLC-LTQ-Orbitrap HRMS. Among them, 59 compounds were confirmed by reference standards. Meanwhile, 21 representative components were simultaneously determined within 15 min by UHPLC-QQQ-MS/MS. 123 components (74 prototypes as well as 49 metabolites) were identified or tentatively characterized. By using network pharmacological analysis, baicalein, liquiritigenin, succinic acid, formononetin, wogonin might be the major effective constituents in QXLZY during the treatment of DN. ConclusionsFlavonoids, saponins and organic acids were the major chemical ingredients of QXLZY. Flavonoids were the main components absorbed into blood, followed by organic acids. Phase II conjugation reaction was the major metabolic type. The pathways that QXLZY in the treatment of DN were probably related to glucose and lipid metabolism, oxidative stress and inflammation.

Lyapunov stability of competitive cells dynamics in tumor mechanobiology

Poromechanics plays a key role in modelling hard and soft tissue behaviours, by providing a thermodynamic framework in which chemo-mechanical mutual interactions among fluid and solid constituents can be consistently rooted, at different scale levels. In this context, how different biological species (including cells, extra-cellular components and chemical metabolites) interplay within complex environments is studied for characterizing the mechanobiology of tumor growth, governed by intratumoral residual stresses that initiate mechanotransductive processes deregulating normal tissue homeostasis and leading to tissue remodelling. Despite the coupling between tumor poroelasticity and interspecific competitive dynamics has recently highlighted how microscopic cells and environment interactions influence growth-associated stresses and tumor pathophysiology, the nonlinear interlacing among biochemical factors and mechanics somehow hindered the possibility of gaining qualitative insights into cells dynamics. Motivated by this, in the present work we recover the linear poroelasticity in order to benefit of a reduced complexity, so first deriving the well-known Lyapunov stability criterion from the thermodynamic dissipation principle and then analysing the stability of the mechanical competition among cells fighting for common space and resources during cancer growth and invasion. At the end, the linear poroelastic model enriched by interspecific dynamics is also exploited to show how growth anisotropy can alter the stress field in spherical tumor masses, by thus indirectly affecting cell mechano-sensing.Graphic

Correlation Analysis of High-Throughput Fruit Phenomics and Biochemical Profiles in Native Peppers (Capsicum spp.) from the Primary Center of Diversification

The main goal of this work was to investigate the relationship between the fruit morphology and biochemical composition of peppers (Capsicum spp.). For that purpose, one hundred native varieties from the Andean region, where the genus Capsicum has its origin, were analysed for different phytochemical compounds. In addition, pepper fruits were assessed with the highly precise phenomics tool Tomato Analyzer. The collection showed a broad variability which was more evident within the C. annuum group. On average, C. frutescens accessions displayed the highest levels of solid soluble content, pH, polyphenols and antioxidant activity. The Tomato Analyzer descriptors under the categories of size, shape index, and latitudinal section, mostly contributed to the variance among Capsicum groups. C. annuum hold the larger fruits, whereas C. frutescens comprised fruits of smaller sizes. The correlation analysis revealed that biochemical traits were negatively correlated with the fruit parameters related to size, suggesting that huger fruits contain lower amounts of chemical metabolites. The multivariate approximations demonstrated that Andean peppers assorted according to morphometric and colorimetric characteristics, but independently of their species or geographical origin. Groups of valuable native varieties carrying promising traits were identified.

Study on taste characteristics and microbial communities in Pingwu Fuzhuan brick tea and the correlation between microbiota composition and chemical metabolites.

The online version of this article contains supplementary material available at (10.1007/s13197-021-04976-y).

English

Capsicum fructescens is a variety of Capsicum with much nutritional and therapeutic usefulness. This study is aimed at investigating the antimicrobial, chemical and biochemical properties of C. fructescens. The biochemical, mineral and phytochemical analyses of the extract were carried out using standard methods. The organic contents of the extracts were determined by GCMS to identify its bioactive constituents which were tested for antibacterial and antifungal activities against 7 bacterial and 3 fungal isolates using disc-diffusion method. The biochemical analysis showed that the leaves were high in protein, fibre, carbohydrate and fat. Different mineral elements were detected in the leaves and they include Magnesium, Calcium, Iron, Sodium, Copper, Potassium, Zinc and Manganese. GC-MS revealed 13 different organic compounds belonging to four groups of chemicals namely alkanol, alkanoic acid, alkanoate and ester. The methanolic extracts of C. fructescens leaves at a dose range of 5 and 25 mg/ml showed significant antibacterial and antifungal activity on some test organisms. The presence of great quantity of dodecanoic acid among other compounds in the extract of bawa suggested the reason for its profound anti-staphylococcal and anti-candidal activities. This study concluded that C. fructescens foliar extract is rich in important chemical and biochemical metabolites which have shown some therapeutic properties. An advocacy in the higher consumption of these peppers among folks is hereby recommended.   Key words: Capsicum fructescens extracts, chemicals and biochemical composition, antimicrobial activity.

Maternal Urinary Phthalate Metabolites are Associated with Lipidomic Profiles Among Pregnant Women in Puerto Rico

Background/Aim: Endocrine disrupting chemicals (EDCs) such as phthalates have been reported to alter circulating lipid concentrations in animals, however, the relationship has rarely been investigated among humans. We assessed associations between phthalate metabolite biomarkers and lipidomic profiles among pregnant women (n = 99) in the Puerto Rico Testsite for Exploring Contamination Threats (PROTECT) birth cohort.Methods: We measured 19 urinary phthalate or phthalate replacement chemical metabolites during 24-28 weeks of pregnancy. Prenatal plasma lipidomic profiles were identified by liquid chromatography-mass spectrometry-based shotgun lipidomics. Multiple statistical strategies were used to assess relationships between phthalates and lipid profiles, including compound-by-compound comparisons using multiple linear regression and various dimension reduction techniques. We derived sums for each lipid class, sums for each lipid sub-class (saturated, monounsaturated, polyunsaturated), and used principal component analysis (PCA) to derive the principal component of each lipid class, which were then regressed on phthalates. Hierarchical clustering and multivariate analyses were also used for further investigation. False discovery rate (FDR) adjusted p-values (q-values) were used to account for multiple comparisons.Results: A total of 587 unique lipids from 19 lipid classes were profiled. When controlling for multiple comparisons, 47 phthalate-lipid associations remained highly significant (p-value<0.001, q-value<0.1). Lipid patterns constructed from PCA accounted for 30~57% of the variation for each lipid class. Mono(2-ethyl-5-carboxypentyl) phthalate (MECCP), mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), and the sum of dibutyl phthalates (ΣDBP) were associated with increased lysophosphatidylcholine (LPC), lysophosphatidylethanolamine (LPE), and phosphatidic acid (PA) lipid groups, particularly those containing saturated and monounsaturated fatty acid chains. This is consistent with the top associations selected from the compound-by-compound analysis. Cyclohexane-1,2-dicarboxylic acid monohydroxy isononyl ester (MHiNCH) was also associated with higher polyunsaturated triglyceride (TG).Conclusion: Certain phthalate/phthalate replacement biomarkers were associated with lipids integral to cell structure and function. Linking these lipid patterns with health parameters will be an important future step.

UHPLC/GC-TOF-MS metabolomics, MTT assay, and molecular docking studies reveal physostigmine as a new anticancer agent from the ethyl acetate and butanol fractions of Kigelia africana (Lam.) Benth. fruit extracts.

Kigelia africana plant is widely used as a herbal remedy in preventing the onset and the treatment of cancer-related infections. With the increase in the research interest of the plant, the specific chemical compound or metabolite that confers its anticancer properties has not been adequately investigated. The ethyl acetate and butanol fractions of the fruit extracts were evaluated by 2-(4,5-dimethylthiazol-2-yl)-3,5-diphenyl-2H-tetrazolium bromide assay against four different cell lines, with the ethyl acetate fraction having inhibition concentration values of 0.53 and 0.42 μM against Hep G2 and HeLa cells, respectively. More than 235 phytoconstituents were profiled using UHPLC-TOF-MS, while more than 15 chemical compounds were identified using GC-MS from the fractions. Molecular docking studies revealed that physostigmine, fluazifop, dexamethasone, sulfisomidine, and desmethylmirtazapine could favorably bind at higher binding energies of -8.3, -8.6, -8.2, and -8.1 kcal/mol, respectively, better than camptothecin with a binding energy of -7.9 kcal/mol. The results of this study showed that physostigmine interacted well with topoisomerase IIα and had a high score of pharmacokinetic prediction using absorption, distribution, metabolism, excretion, and toxicity profiles, thereby suggesting that drug design using physostigmine as a base structure could serve as an alternative against the toxic side effects of doxorubicin and camptothecin.

Partial least squares regression with compositional response variables and covariates

ABSTRACT The common approach for regression analysis with compositional variables is to express compositions in log-ratio coordinates (coefficients) and then perform standard statistical processing in real space. Similar to working in real space, the problem is that the standard least squares regression fails when the number of parts of all compositional covariates is higher than the number of observations. The aim of this study is to analyze in detail the partial least squares (PLS) regression which can deal with this problem. In this paper, we focus on the PLS regression between more than one compositional response variable and more than one compositional covariate. First, we give the PLS regression model with log-ratio coordinates of compositional variables, then we express the PLS model directly in the simplex. We also prove that the PLS model is invariant under the change of coordinate system, such as the ilr coordinates with a different contrast matrix or the clr coefficients. Moreover, we give the estimation and inference for parameters in PLS model. Finally, the PLS model with clr coefficients is used to analyze the relationship between the chemical metabolites of Astragali Radix and the plasma metabolites of rat after giving Astragali Radix.

Rapid identification of chemical composition and metabolites of Pingxiao Capsule in vivo using molecular networking and untargeted data-dependent tandem mass spectrometry.

Pingxiao capsule (PXC) is a herbal medicine used for adjuvant therapy in breast cancer. However, the constituents and absorbed components of the formula and their related metabolites have not been elucidated to date. PXC is a typical traditional Chinese medicine formula consisting of Strychnos nux-vomica L., Curcuma wenyujin Y. H., Agrimonia pilosa Ledeb., Toxicodendron vernicifluum, Trogopterus dung, alumen, potassium nitrate (saltpeter) and Citrus aurantium L. In this study, a ultra-high performance liquid chromatography system equipped with high resolution Q-Orbitrap mass spectrometry (MS) and comparative Global Natural Product Social molecular networking together with the Compound Discoverer software were used to identify metabolites of PXC in vitro and in vivo. Based on untargeted data-dependent MS2 and data-mining techniques, 89 peaks of alkaloids, flavonoids, organic acid and phenolic compounds were identified in a PXC 70% methanol extract. Furthermore, 15 absorbed prototype compounds and their metabolites were rapidly confirmed in rat blood. Glucuronidation, oxidation, methylation and hydroxylation were the main metabolic pathways. We fully clarified the chemical constituents of PXC and provided a scientific and efficient strategy for rapid discovery and identification of prototypes and their metabolites in rat plasma using high-resolution MS aided by Global Natural Product Social and Compound Discoverer software.

1H NMR-Based Metabolomics Coupled With Molecular Docking Reveal the Anti-Diabetic Effects and Potential Active Components of Berberis vernae on Type 2 Diabetic Rats.

The dried stem bark of Berberis vernae C.K.Schneid., known as “Xiao-bo-pi” in Chinese, is a representative anti-diabetic herb in traditional Tibetan medical system. However, its anti-diabetic mechanisms and active components remain unclear. In this study, 1H NMR-based metabolomics, biochemistry assay, molecular docking, and network analysis were integrated to evaluate the anti-diabetic effects of B. vernae extract on type 2 diabetic rats, and to explore its active components and underlying mechanisms. Diabetes was induced by high-fat diet and streptozotocin. After 30 days of treatment, B. vernae extract significantly decreased the serum levels of fasting blood glucose, insulin, insulin resistance index, glycated serum protein, TNF-α, IL-1β, and IL-6, whereas significantly increased the serum levels of insulin sensitivity index in type 2 diabetic rats. A total of 28 endogenous metabolites were identified by 1H NMR-based metabolomics, of which 9 metabolites that were changed by diabetes were significantly reversed by B. vernae extract. The constructed compound-protein-metabolite-disease (CPMD) interaction network revealed the correlation between chemical constituents, target proteins, differential metabolites, and type 2 diabetes. Ferulic acid 4-O-β-D-glucopyranoside, bufotenidine, jatrorrhizine, and berberine showed good hit rates for both the 30 disease-related proteins and 14 differential metabolites-related proteins, indicating that these four compounds might be the active ingredients of B. vernae against type 2 diabetes. Moreover, pathway analysis revealed that the anti-diabetic mechanisms of B. vernae might be related to its regulation of several metabolic pathways (e.g., butanoate metabolism) and disease-related signal pathways (e.g., adipocytokine signaling pathway). In summary, B. vernae exerts a significant anti-diabetic effect and has potential as a drug candidate for the treatment of type 2 diabetes.

Insights into the development of hepatocellular fibrillar inclusions in European flounder (Platichthys flesus) from UK estuaries

Hepatocellular fibrillar inclusions (HFI) are an unusual pathology of unknown aetiology affecting European flounder (Platichthys flesus), particularly from estuaries historically impacted by pollution. This study demonstrated that the HFI prevalence range was 6–77% at several UK estuaries, with Spearman rank correlation analysis showing a correlation between HFI prevalence and sediment concentrations of ∑PBDEs and ∑HBCDs. The data showed that males exhibit higher HFI prevalence than females, with severity being more pronounced in estuaries exhibiting higher prevalence. HFI were not age associated indicating a subacute condition. Electron microscopy confirmed that HFI were modified proliferating rough endoplasmic reticulum (RER), whilst immunohistochemistry provided evidence of VTG production in HFI of male P. flesus. Despite positive labelling of aberrant VTG production, we could not provide additional evidence of xenoestrogen exposure. Gene transcripts (VTG/CHR) and plasma VTG concentrations (>1 μg ml−1), were only considered elevated in four male fish showing no correlation with HFI severity. Further analysis revealed that reproductively mature female P. flesus i.e. >3-year-old, did not exhibit HFI, whereas males of all ages were affected. This, combined with previous reports that estradiol (E2) can impair mixed function oxygenase activity, supports a hypothesis that harmful chemical metabolites (following phase 1 metabolism of their parent compounds) are potentially responsible for HFIs observed in male and ≤ 3-year-old female fish. Consequently, HFI and xenoestrogenic induced VTG production could be independent of each other resulting from different concurrent toxicopathic mechanisms, although laboratory exposures will likely be the only way to determine the true aetiology of HFI.

Identification of Metabolic Alterations in Breast Cancer Using Mass Spectrometry-Based Metabolomic Analysis.

Breast cancer (BC) is a major global health issue and remains the second leading cause of cancer-related death in women, contributing to approximately 41,760 deaths annually. BC is caused by a combination of genetic and environmental factors. Although various molecular diagnostic tools have been developed to improve diagnosis of BC in the clinical setting, better detection tools for earlier diagnosis can improve survival rates. Given that altered metabolism is a characteristic feature of BC, we aimed to understand the comparative metabolic differences between BC and healthy controls. Metabolomics, the study of metabolism, can provide incredible insight and create useful tools for identifying potential BC biomarkers. In this study, we applied two analytical mass spectrometry (MS) platforms, including hydrophilic interaction chromatography (HILIC) and gas chromatography (GC), to generate BC-associated metabolic profiles using breast tissue from BC patients. These metabolites were further analyzed to identify differentially expressed metabolites in BC and their associated metabolic networks. Additionally, Chemical Similarity Enrichment Analysis (ChemRICH), MetaMapp, and Metabolite Set Enrichment Analysis (MSEA) identified significantly enriched clusters and networks in BC tissues. Since metabolomic signatures hold significant promise in the clinical setting, more effort should be placed on validating potential BC biomarkers based on identifying altered metabolomes.

Chemical defense against microfouling by allelopathic active metabolites of Halymenia floresii (Rhodophyta)

During the experimental cultivation of the red alga Halymenia floresii under Integrated MultiTrophic Aquaculture (IMTA), the establishment of opportunist green algae and the colonization of sessile invertebrates, which were usually disturbing the cultivation, were not observed. The culture tanks were clean and the surface of the H. floresii was remarkably free from any fouling organisms. This phenomenon could reveal that the presence of H. floresii may prevent biofilm formation by releasing allelopathic active compounds that ultimately interfere with the settlement and growth of competitors. In order to understand this phenomenon, H. floresii was cultivated under controlled environmental conditions and analyzed for its surface chemical defense metabolites. The surface-associated metabolites were extracted using the DIP extraction method, using different solvents with increasing polarity and immersion periods. Using epifluorescence microscopy, n-hexane was found to be the suitable immersion solvent for H. floresii for a period of 10 to 60 s to extract surface metabolites. The whole cell metabolites were extracted exhaustively with the same solvent for a period of 24 h. The chemical profiling of the surface compounds was performed by liquid chromatography mass spectroscopy (LC-MS), followed by a Mass Bank search and compared with those obtained from the whole-cell extracts. The mean concentration of H. floresii surface metabolites was 600 ng cm−2 (c. 60 g of a fresh sample) whereas the whole-cell metabolite concentration was around 4.5 μg mg−1 (400 mg of the lyophilized sample). The bioactivity of the H. floresii surface extracts was studied by evaluating their quorum quenching behavior on the surface-associated bacteria. The cultivable bacteria isolated from the surface of H. floresii were identified as Vibrio owensii (B3IM), Alteromonas sp. (B7CC), Pseudoalteromonas arabiensis (B4BC), Ruegeria sp. (B4CC), Tenacibaculum sp.(B9BC), Maribacter sp. (B9IM) and Aquimarina sp. (B9.1CC). All the isolated strains belonged to Alphaproteobacteria, Gammaproteobacteria and Bacteroides. The results of this bioactivity proved that the surface-associated metabolite extract (DIP) interferes with the communication signals produced by the bacteria isolates with the reporter strain employed. According to the Mass Bank compound analysis, we hypothesized that flavonoids and/or halogenated compounds might have contributed to this activity. This work provides an understanding of the influence of surface-associated metabolites on the associated bacterial community and by which H. floresii manages to control the biofouling on its surface.

Design and Rationale of the Biomarker Center of the Household Air Pollution Intervention Network (HAPIN) Trial.

Background:Biomarkers of exposure, susceptibility, and effect are fundamental for understanding environmental exposures, mechanistic pathways of effect, and monitoring early adverse outcomes. To date, no study has comprehensively evaluated a large suite and variety of biomarkers in household air pollution (HAP) studies in concert with exposure and outcome data. The Household Air Pollution Intervention Network (HAPIN) trial is a liquified petroleum gas (LPG) fuel/stove randomized intervention trial enrolling 800 pregnant women in each of four countries (i.e., Peru, Guatemala, Rwanda, and India). Their offspring will be followed from birth through 12 months of age to evaluate the role of pre- and postnatal exposure to HAP from biomass burning cookstoves in the control arm and LPG stoves in the intervention arm on growth and respiratory outcomes. In addition, up to 200 older adult women per site are being recruited in the same households to evaluate indicators of cardiopulmonary, metabolic, and cancer outcomes.Objectives:Here we describe the rationale and ultimate design of a comprehensive biomarker plan to enable us to explore more fully how exposure is related to disease outcome.Methods:HAPIN enrollment and data collection began in May 2018 and will continue through August 2021. As a part of data collection, dried blood spot (DBS) and urine samples are being collected three times during pregnancy in pregnant women and older adult women. DBS are collected at birth for the child. DBS and urine samples are being collected from the older adult women and children three times throughout the child’s first year of life. Exposure biomarkers that will be longitudinally measured in all participants include urinary hydroxy-polycyclic aromatic hydrocarbons, volatile organic chemical metabolites, metals/metalloids, levoglucosan, and cotinine. Biomarkers of effect, including inflammation, endothelial and oxidative stress biomarkers, lung cancer markers, and other clinically relevant measures will be analyzed in urine, DBS, or blood products from the older adult women. Similarly, genomic/epigenetic markers, microbiome, and metabolomics will be measured in older adult women samples.Discussion:Our study design will yield a wealth of biomarker data to evaluate, in great detail, the link between exposures and health outcomes. In addition, our design is comprehensive and innovative by including cutting-edge measures such as metabolomics and epigenetics. https://doi.org/10.1289/EHP5751

Isolation, identification and antimicrobial activity of secondary metabolites of endophytic fungi from annona leaves (Annona squamosa L.) growing in dry land

Research on isolation, identification, and antimicrobial activity of secondary metabolites of endophytic fungi from leaves of Annona squamosa growing in dryland had been undertaken. The work includes the isolation of endophytic fungi, cultivation, and extraction of fungal extract and identification of chemical metabolites together with the antibacterial test. The pure colony of endophytic fungi was grown on solid rice media using 1 L Erlenmeyer flask. The grown fungi were extracted with ethyl acetate. The ethyl acetate crude extract was then further subjected to chemical analysis and its antibacterial properties. Endophytic fungi species was identified as Aspergillus niger based on macroscopic and microscopic analysis. Analysis LC-MS/MS had revealed the presence of five metabolites including ephedradine A, ergosine, Ia, mudanpioside H, and trichosanic acid. The extract showed strong inhibition against Staphyloccus aureus with diameter of zone inhibition of 16.1 mm and moderate inhibition against Escherichia coli 0175H7 and Salmonella enteritidis ATCC 6939 with the observed diameter of zone inhibition of 9.6 mm and 11.3 mm, respectively.

Antimicrobial, antioxidant, phytochemical and pharmacognostic study of the leaf powder of Ficus carica L.

Ficus carica L., (Moraceae), commonly found in tropical and subtropical areas, known for its ethno-medicinal uses. The current work aimed to study macro and microscopic features and screen the plant ethanol extract for its antioxidant and antimicrobial potential. 1, 1-diphenyl-2-picryl hydrazyl (DPPH) was used for antioxidant and Agar-well-diffusion method for screening antimicrobial activity. The results revealed that powdered drug possess unicellular, uniserrate covering trichomes, parenchymatous tissue containing spiral vascular strands, anomocytic stomata and calcium oxalate crystals. Phytochemically, the extract possesses reducing sugars, polysaccharides, oxalic acid, amino acid and proteins. The antioxidant activity of Ficus carica leaves ethanol extract, using various concentration (125, 250, 500, 750 and 1000 µg/ml) gave the results of 21.42±01, 29.65±03, 53.15±03, 57.00±01 and 62.99±05, respectively. The leaves ethanol extract used for antibacterial activity have concentrations of 200 and 500 mg/ml, both of which were found effective against the selected bacterial strains. K. pneumonia was inhibited (18 & 28mm), E. coli was found (20 & 26 mm) susceptible, Staphylococcus aureus was inhibited with (24 and 26mm) and Pseudomonas aeruginosa was inhibited by (22 and 28mm). It is concluded from the present study that leaf ethanol extract of Ficus carica has good antioxidant constituents and possess positive antimicrobial chemical metabolites. These metabolites can be used to treat various infectious and chronic diseases caused by microbes. This work is important in order to assess the significance of Ficus carica used locally. More research is required to isolate these metabolites to establish clinical trials for human benefits. Keywords: Antimicrobial; Antioxidant; Ficus Carica; Medicinal Plants; Phytochemicals http://dx.doi.org/10.19045/bspab.2020.90105

Characterization of chemical components of Periplocae Cortex and their metabolites in rats using ultra‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry

Periplocae Cortex, named Xiang-Jia-Pi in China, has been widely used to treat autoimmune diseases, especially rheumatoid arthritis. However, the in vivo substances of Periplocae Cortex remain unknown yet. In this study, an ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used for profiling the chemical components and related metabolites of Periplocae Cortex. A total of 98 constituents were identified or tentatively characterized in Periplocae Cortex: 42 C21 steroidal glycosides, 10 cardiac glycosides, 23 organic acids, 4 aldehydes, 7 triterpenes, and 12 other types. Among them, 18 components were unambiguously identified by comparison with reference standards. In addition, 176 related xenobiotics (34 prototypes and 142 metabolites) were screened out and characterized in rats' biosamples (plasma, urine, bile, and feces) after the oral administration of Periplocae Cortex. Moreover, the metabolic fate of periplocoside S-4a, a C21 steroidal glycoside, was proposed for the first time. In summary, phase II reactions (methylation, glucuronidation, and sulfation), phase I reactions (hydrolysis reactions, oxygenation, and reduction), and their combinations were the predominant metabolic reactions of Periplocae Cortex in rat. It is the first report to reveal the in vivo substances and metabolism feature of Periplocae Cortex. This study also provided meaningful information for further pharmacodynamics study of Periplocae Cortex, as well as its quality control research.

Delivery of microbial metabolites for human health care: a review

Microbial bioactive compounds are one of the most natural and raw form of chemical metabolites present in nature. There has been recently an increasing interest in the use of microorganisms for the isolation and production of metabolites for application in health care. A large number of microbial metabolites have been isolated and screened for the treatment of human diseases. Here, we review microbial isolates having antidiabetic, anticancerous, antibacterial and antifungal properties. The advanced automated equipments have allowed to screen a large number of microbial compounds in vitro. Yet there are still major challenges for the delivery of bioactive compounds in vivo. Here, we focus on microbial metabolites used as therapeutics, and delivery routes of microbial metabolites in vivo.

Characterization of covalent protein modification by triclosan in vivo and in vitro via three-dimensional liquid chromatography-mass spectrometry: New insight into its adverse effects

Triclosan (TCS), an antimicrobial agent widely used in personal care products and ubiquitously exists in environment, has drawn increasing concern due to its potential to exert multiple adverse effects, ranging from endocrine disruption to carcinogenesis. However, the mechanism of these adverse effects is still not fully elucidated. More and more studies have shown that chemical reactive metabolites (RMs) covalently binding to proteins is a possible reason for these adverse effects, but there is still a lack of appropriate methods to predict or evaluate these adverse effects due to the extremely low abundance of the modified proteins in complex biological samples. In this study, we attempted to address this problem and investigate the possible mechanism of TCS adverse effects by a shotgun proteomics approach based on three-dimensional-liquid chromatography-mass spectrometry (3D-LC-MS). First, the in vitro incubation with model amino acids and protein in microsomes showed that TCS could react with cysteine residue of proteins through 3 types of RMs. Then, a 3D-LC-MS approach was developed to sensitively determine the low abundant modified proteins, which resulted in the identification of 45 TCS-modified proteins, including albumin, haptoglobin and NR1I2, in rats. STRING analysis indicated that these modified proteins mainly were involved in reproductive and development system, endocrine and immune system, and carcinogenesis, which were in accord with the main reported TCS-induced adverse effects and suggested that the covalent modification of TCS RMs for proteins might affect their activities and functions, thus inducing serious adverse effects. This study provided a new insight into the mechanism of TCS adverse effects and may serve as a valuable method to predict or evaluate adverse effects of ubiquitous chemicals.

Comprehensive investigation of mechanism and effective ingredients of Fangji Huangqi Tang by serum pharmacochemistry and network pharmacology.

Fangji Huangqi Tang (FHT), has been reported to show effects on nephrotic syndrome, but its mechanism of action and bioactive components have not yet been determined. In this study, a method using UPLC-HRMS/MS was established for the detection and identification of the chemical constituents and metabolites absorbed into the blood. Absorbed components in serum were then used for the network pharmacology analysis to deduce the mechanism and effective components. A total of 86 compounds were identified or tentatively characterized. Based on the same instrumental conditions, 85 compounds were found in rat serum after oral administration of FHT, including 22 prototypes and 63 metabolites. Network pharmacology analysis showed that absorbed components, such as (3R)-2',3',4',7-tetrahydroxyisoflavan, astrapterocarpan, cycloastragenol, 7,2'-dihydroxy-3',4'-dimethoxyisoflavan, astragaloside IV, astrapterocarpan glucoside and glycyrrhetinic acid, could be responsible for the pharmacological activity of nephrotic syndrome by regulating the VEGF signaling pathway, focal adhesion and MAPK signaling pathway. Furthermore, the pathway-target network showed that the MAPK1, AKT2 and CDC42 were involved in the signal pathways above. This study provides a scientific basis for the mechanism and effective ingredients of FHT.