Comprehensive characterization of the chemical constituents of Lianhua Qingwen capsule by ultra high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry.

Lianhua Qingwen capsule is a famous traditional Chinese medicine (TCM) prescription that is widely used for the treatment of respiratory diseases in China. To facilitate in-depth and global characterization of the chemical constituents of Lianhua Qingwen capsule, a profiling method based on ultra-high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) was applied in both positive and negative ion modes for the comprehensive characterization of the chemical profiles of Lianhua Qingwen capsule. A total of 596 compounds were identified or tentatively characterized, including 137 flavonoids, 46 phenylpropanoids, 43 phenylethanoid glycosides, 145 terpenoids, 83 organic acids and their derivatives, 15 quinones, 39 alkaloids, 32 alcohol glycosides and 56 other compounds. Thus, this results widely extended and enriched the chemical constituents of Lianhua Qingwen capsule, which will provide comprehensive and valuable information for its quality control and further pharmacological study, facilitate understanding the effective substance and pharmacodynamic material basis, thereby providing a solid foundation for further development of the Lianhuaqingwen capsule.

Deciphering the enigma between low bioavailability and high anti-hepatic fibrosis efficacy of Yinchen Wuling powder based on drug metabolism and network pharmacology.

ETHNOPHARMACOLOGICAL RELEVANCE: Yinchen Wuling powder (YCWLP) is a famous traditional Chinese medicine formula with the effect of "removing jaundice and eliminating dampness", which has the potential to prevent and treat hepatic fibrosis (HF). However, the mechanism of the active ingredients of YCWLP in treating HF remains to be clarified. AIM OF THE STUDY: This study aims to investigate the in vivo metabolic profile of YCWLP and the mechanism of its gut microbiota-mediated therapeutic effect on HF via network pharmacology. MATERIALS AND METHODS: In this comprehensive study, the UHPLC-FT-ICR-MS platform was used for the systematic characterization of the in vivo metabolic profile of YCWLP, and the mediating effect of gut microbiota was elucidated by comparing the differences of metabolites between the normal rats and pseudo germ-free rats administrated with YCWLP. Then, the identified active ingredients of YCWLP metabolized by gut microbiota and their targets associated with HF were used for further network pharmacological analysis, including the construction of PPI network, GO and KEGG enrichment and compound-target-pathway-disease network. RESULTS: Overall, 41 prototype compounds and 138 metabolites were identified in the biosamples after YCWLP administration. Among them, 15 drug prototypes are clearly metabolized by gut microbiota, and 91 metabolites showed significant differences between the N-YCWLP group and the PGF-YCWLP group, which might be attributed to the mediation of gut microbiota. Network pharmacology studies on the aforementioned 15 prototype components indicated crucial roles of arginine biosynthesis and complement and coagulation cascades-related genes such as PLG, NOS3, GC and F2 in the treatment of HF by YCWLP mediated by gut microbiota. CONCLUSIONS: The therapeutic effects of multiple active ingredients in YCWLP on HF depend on the metabolism of gut microbiota. This study offers novel insights into the relationship between bioactive chemical constituents and the action mechanism of YCWLP against HF.

Systematic Screening of the Chemical Constituents of Lanqin Oral Liquid by Ultra-High-Performance Liquid Chromatography Combined with Fourier Transform Ion Cyclotron Resonance Mass Spectrometry.

A rapid and sensitive method that combined ultra-high-performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) was used to identify the chemical constituents in Lanqin oral liquid. On the basis of UHPLC-FT-ICR-MS analysis, systematic characterization of the chemical profile of Lanqin oral liquid was carried out, and a total of 441 compounds were identified or tentatively characterized including alkaloids, flavonoids, terpenoids, organic acids, phenylpropanoids, and other types. The results provide a reference for improving quality control, contribute to establishing higher quality standards, provide a scientific basis for further research on the pharmacodynamic material basis, and help illustrate the relationship between the complicated constituents and therapeutic effects of Lanqin oral liquid.

Determination of the components of danyikangtai powder into the plasma and its pharmacodynamic study.

Danyikangtai powder has a definite therapeutic effect on pancreatitis. However, the internal mechanism is unclear. The purpose of this experiment is to quickly identify the blood components of danyikangtai powder and evaluate its efficacy. 25 blood components were identified by comparing the components with the same mass spectrometry information from in vivo and in vitro samples. The AR42J cells of the pancreatitis model were treated with drug-containing plasma, and the drug efficacy was evaluated by investigating the amylase release rate. This study provides a scientific reference for its pharmacological research and rational clinical application.

Screening of differential components of Gegenqinlian decoction and their comparative pharmacokinetics in normal and pyrexia rats using UHPLC-FT-ICR-MS and UHPLC-MS/MS.

UHPLC combined with Fourier-transform ion cyclotron resonance MS metabonomic approach was employed to screen the differential components between normal rats and yeast-induced pyrexia rats after an oral administration of Gegenqinlian decoction (GQLD). Nine compounds, namely puerarin, daidzein, baicalin, wogonoside, wogonin, berberine, palmatine, jateorhizine, and coptisine, were identified as differential components in the plasma. A rapid, sensitive, selective, and accurate UHPLC-MS method was developed and fully validated for the simultaneous determination of the screened components in rat plasma after an oral administration of GQLD. The values for the limit of quantification ranged from 0.025 to 5.0 ng/mL. The inter- and intra-day precision of all analytes was ≤10.7%, with an accuracy of ≤10.5%. Good extraction recovery and matrix effects were also obtained. The method was successfully applied to a comparative pharmacokinetic study of GQLD in normal and pyrexia rats. The results showed that the pharmacokinetic behavior of the analytes was changed in pyrexia rats compared to normal rats. These results could provide beneficial guidance for clinical applications of GQLD.

Rapid characterization of chemical constituents of Gansuibanxia decoction by UHPLC-FT-ICR-MS analysis.

Gansuibanxia decoction (GSBXD) is one of the most famous traditional Chinese medicine (TCM). It is a herbal formula used for treating hydrops, such as cancerous ascites, pleural effusion, pericardial effusion, etc. However, the chemical constituents of GSBXD were still unclear. In this study, an UHPLC-FT-ICR-MS method was established and applied to the separation and characterization of the chemical constituents of GSBXD. A total of 62 components were chemically defined or tentatively identified, including diterpenoids, triterpenoids, flavonoids, monoterpene glycosides and alkaloids. The results is meaningful for a better understanding of the material basis of GSBXD and can be the basis for its further in vitro and in vivo studies.

Metabonomic analysis of plasma biochemical changes in pyrexia rats after treatment with Gegenqinlian decoction, aspirin and itraconazole by UHPLC-FT-ICR-MS / 药物分析学报

A metabonomic approach involving an ultrahigh-performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) was used to investi-gate the changes in the endogenous metabolites in the plasma of rats with yeast-induced pyrexia treated with Gegenqinlian decoction (GQLD), aspirin and itraconazole. The differences in the small molecule profiles of treatment using traditional Chinese medicine, etiological treatment and symptomatic treat-ment were elucidated. Thirty-six plasma metabolites were identified or putatively identified, and the effects of the three medicines on the thirty-six metabolites were studied. Their metabolic pathways indicated that GQLD, aspirin and itraconazole ameliorated the rats with yeast-induced pyrexia pre-dominantly by regulating the metabolisms of phospholipid, sphingolipid, fatty acid oxidation, fatty acid amides, amino acid and glycerolipid in vivo. The pharmacodynamics and metabonomic results showed that the three medicines exhibited the therapeutic effects on pyrexia by regulating the perturbations of multiple metabolisms. The study provided a scientific basis for an in-depth understanding of the ther-apeutic effects of GQLD, aspirin and itraconazole on rats with yeast-induced pyrexia.

Metabonomic analysis of plasma biochemical changes in pyrexia rats after treatment with Gegenqinlian decoction, aspirin and itraconazole by UHPLC-FT-ICR-MS.

A metabonomic approach involving an ultrahigh-performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) was used to investigate the changes in the endogenous metabolites in the plasma of rats with yeast-induced pyrexia treated with Gegenqinlian decoction (GQLD), aspirin and itraconazole. The differences in the small molecule profiles of treatment using traditional Chinese medicine, etiological treatment and symptomatic treatment were elucidated. Thirty-six plasma metabolites were identified or putatively identified, and the effects of the three medicines on the thirty-six metabolites were studied. Their metabolic pathways indicated that GQLD, aspirin and itraconazole ameliorated the rats with yeast-induced pyrexia predominantly by regulating the metabolisms of phospholipid, sphingolipid, fatty acid oxidation, fatty acid amides, amino acid and glycerolipid in vivo. The pharmacodynamics and metabonomic results showed that the three medicines exhibited the therapeutic effects on pyrexia by regulating the perturbations of multiple metabolisms. The study provided a scientific basis for an in-depth understanding of the therapeutic effects of GQLD, aspirin and itraconazole on rats with yeast-induced pyrexia.

Investigation of the mechanism of incompatible herb pair gansui-gancao-induced hepatotoxicity and nephrotoxicity and the attenuated effect of gansuibanxia decoction by UHPLC-FT-ICR-MS-based plasma metabonomic analysis.

Gansui-Gancao is one of the "eighteen incompatible herb pairs" which was recorded 2000 years ago according to TCM (Traditional Chinese Medicine) theory for their toxicity when using together. Nevertheless, Gansuibanxia decoction contained the herb pair have satisfactory effect on the treatment of cancerous ascites, pericardial effusion, etc. The present study aimed to investigate the mechanism of the incompatibility of Gansui-Gancao and the compatibility of Gansuibanxia decoction using UHPLC-FT-ICR-MS in a metabonomic perspective. Rats were divided into four groups administrated with different herb combination extracts for successive 14 days. Orthogonal partial least squares-discriminant analysis (OPLS-DA) was used to plot the metabolic state and screen the potential biomarkers in plasma. A total of 20 biomarkers contributed to the separation of Gansui-Gancao group and control group were tentatively identified mainly involved in 7 metabolic pathways related to hepatotoxicity and nephrotoxicity. The contents of these biomarkers were adjusted to normal levels in Gansuibanxia decoction group. Thus, the results of our study reveled the mechanism of the incompatibility of Gansui-Gancao and the compatibility of Gansuibanxia decoction in a metabonomic perspective and it's valuable for better understanding the "eighteen incompatible madicaments" of TCM theory.

Metabolic profiling of Gegenqinlian decoction in rat plasma, urine, bile and feces after oral administration by ultra high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry.

Gegenqinlian decoction (GQLD) as a traditional Chinese medical (TCM) prescription is made from four Chinese herbs and has been used for centuries to treat pyrexia and diarrhea. In this work, an ultra-high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) method was established to profile the metabolic fate of GQLD in rat plasma, urine, bile and feces. The analysis was performed on a Waters Acquity UPLC HSS T3 column (2.1 mm × 100 mm, 1.8 µm) at 35 °C within 25 min. A total of 174 compounds mainly including flavonoids, alkaloids and triterpenoids in rat plasma, urine, bile and feces after oral administration of GQLD were identified or tentatively characterized by comparison with retention times, accurate mass within 5 ppm error and their characteristic MS fragmentation ions. Among compounds in rats, 107 compounds are prototypes and 67 compounds are metabolites. Results demonstrated that metabolic pathways of GQLD in rats included methylation, demethylation, hydrogenation, hydrolysis, hydroxylation, methoxylation, sulfate conjugation and glucuronide conjugation. A clear understanding of the absorption and metabolism of GQLD is very important in its rational clinical use and pharmacological research. These findings enhance our understanding of the metabolism and effective forms of GQLD.

Characterization of global metabolic profile of Rhodiola crenulata after oral administration in rat plasma, urine, bile and feces based on UHPLC-FT-ICR MS.

Rhodiola crenulata has been widely used as a health food, antifatigue and antidepressant in China and many other countries for centuries. However, to date the metabolism of it in vivo still remains unclear. In this study, UHPLC-FT-ICR MS was used to analyze the major components and their metabolites in rats after oral administration of Rhodiola crenulata for the first time. A total of 179 constituents, including 37 prototype compounds and 142 metabolites (89 phase I metabolites and 53 phase II metabolites) were tentatively identified. The metabolic pathways included hydroxylation, deglycosylation, dehydrogenation, glucuronidation and sulphate conjugation. In summary, this study showed an insight into the metabolism of Rhodiola crenulata in vivo, which may provide helpful chemical information for better understanding the multiple functions of it. And also, the developed method could be used as a reliable strategy to study the metabolic profile for other traditional chinese medicines.

In vivo study of erysolin metabolic profile by ultra high performance liquid chromatography coupleded to Fourier transform ion cyclotron resonance mass spectrometry.

An ultra high performance liquid chromatography coupled to Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) method was developed for the first time to study the in vivo metabolism of erysolin, a compound derived from cruciferous plants which has a definite effect of anti-tumor and anti-nerve injury. In this research, the chromatographic separation was performed on an ACQUITY UPLC® BEH C18 column (2.1 mm×100mm, 1.7µm, Waters, USA) and eluted by a gradient program, the identification work was achieved on a Bruker ultra-high resolution spectrometer in positive ion mode. Plasma, urine, feces and bile samples were collected from rats to screen metabolites after an intragastric administration of erysolin at the dose of 100mg/kg. As a result, the parent drug and a total of six phase II metabolites were detected and preliminarily identified by analyzing their MS and MS/MS spectrometry profiles. Our results indicated that erysolin mainly metabolized via the mercapturic acid metabolic pathway, erysolin first react with glutathione to form glutathione conjugate, followed by taking off the glutamic acid and glycine to form cysteine conjugate, then the N-acetylation reaction occurs, the product would be excreted out of the body at last. In conclusion, results obtained in our study may contribute to a better understanding of the metabolism process and characteristics of erysolin in vivo, and provide an important reference for future research.

A plasma metabonomic analysis on potential biomarker in pyrexia induced by three methods using ultra high performance liquid chromatography coupled with Fourier transform ion cyclotron resonance mass spectrometry.

Pyrexia usually is a systemic pathological process that can lead to metabolic disorders. Metabonomics as a powerful tool not only can reveal the pathological mechanisms, but also can give insight into the progression of pyrexia from another angle. Thus, an ultra high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) metabonomic approach was employed for the first time to investigate the plasma biochemical characteristics of pyrexia induced by three methods and to reveal subtle metabolic changes under the condition of pyrexia so as to explore its mechanism. The acquired metabolic data of the models were subjected to principal component analysis (PCA) for allowing the clear separation of the pyrexia rats from the control rats. Variable importance for project values (VIP) and Student's t-test were used to screen the significant metabolic changes caused by pyrexia. Fifty-two endogenous metabolites were identified and putatively identified as potential biomarkers primarily associated with phospholipid metabolism, sphingolipid metabolism, fatty acid oxidation metabolism, fatty acid amides metabolism and amino acid metabolism, and related to bile acid biosynthesis and glycerolipid catabolism. LysoPC (14:0), LysoPC (18:3), LysoPC (20:4), LysoPC (16:0), phytosphingosine, Cer (d18:0/12:0), N-[(4E,8E)-1,3-dihydroxyoctadeca-4,8-dien-2-yl]hexadecanamide, oleamide, fatty acid amide C22:1, tryptophan, acetylcarnitine, palmitoylcarnitine and stearoylcarnitine were considered as common potential biomarkers of pyrexia rats induced by three methods: Our results revealed that the UHPLC-FT-ICR-MS-based metabolomic method is helpful for finding new potential metabolic markers for pyrexia detection and offers a good perspective in pyrexia research.

A rapid and sensitive UHPLC-FT-ICR MS/MS method for identification of chemical constituents in Rhodiola crenulata extract, rat plasma and rat brain after oral administration.

A rapid and sensitive UHPLC-FT-ICR MS/MS method was developed for the first time to analyze the extract of Rhodiola crenulata and the constituents absorbed into rat blood and brain after oral administration. Under the optimized conditions, a total of 64 chemical constituents were identified or tentatively characterized in vitro in 30min, and also 24 and 9 chemical constituents were detected in rat plasma and brain respectively, by comparing the retention time, accurate mass and/or MS/MS data of blank and dosed sample. The results indicated that the developed UHPLC-FT-ICR MS/MS method was suitable for detection and identifying the chemical constituents in Rhodiola crenulata extract, rat plasma and rat brain, and it could be used as a powerful and reliable analytical strategy for rapid identification of chemical constituents in vitro and in vivo for other traditional Chinese herbal medicines (TCMs). Furthermore, the detected chemical constituents in rat brain could be speculated to be the pharmacodynamic substances of Rhodiola crenulata for Alzheimer's disease (AD) and it could also provide useful chemical information for further mass spectrometry imaging and bioactive substances research on Rhodiola crenulata.

Metabolic profile of 2-(2-hydroxypropanamido) benzoic acid in rats by ultra high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry.

An ultra high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry (UHPLC-FT-ICR-MS) method was developed to investigate the in vivo metabolism of 2-(2-hydroxypropanamido) benzoic acid (HPABA), a marine-derived anti-inflammatory drug, for the first time. Plasma, urine, feces and bile samples were collected from male and female rats after a single intragastric administration of HPABA at a dose of 100mg/kg. Besides the parent drug, a total of 13 metabolites (3 phase I and 10 phase II metabolites) were detected and tentatively identified through comparing their mass spectrometry profiles with those of HPABA. Results demonstrated that metabolic pathways of HPABA in rats included decarboxylation, hydroxylation, dehydrogenation, glucuronidation, glycine conjugation and N-acetyl conjugation. In summary, this work provided valuable information regarding the metabolism of HPABA in rats, which would contribute to better understanding of its safety and mechanism of action.