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ABSTRACT Identification of multicomponent in traditional Chinese medicine (TCM) is complex and time-consuming. The inspection of the full-scan mass chromatograms was usually performed manually, which is labor-intensive. It is difficult [...]

Present studies have shown that Flos Chrysanthemi has anti-inflammatory and other effects and regulates intestinal function, while the chrysanthemum stem and leaf as non-medicinal parts of chrysanthemum have similar chemical components with chrysanthemum, but the activity and mechanisms are rarely elucidated. Therefore, this study used a DSS-induced zebrafish inflammatory bowel disease model to study the anti-inflammatory and antioxidant effects of chrysanthemum stem and leaf extracts. The results indicate that DSS induction leads to increased secretion of acidic mucin in the intestines of juvenile fish, enlargement of the intestinal lumen and the emergence of intestinal inflammation. Compared with the model group, each administration group differentially inhibited the expression of IL-1β, IL-8 and MMP9 in DSS-induced zebrafish, while upregulating the activity of superoxide dismutase. The quantitative analysis results showed that the flavonoids (including Linarin, Diosmetin-7-glucoside, Tilianin, etc.) and phenolic acids (including Isochlorogenic acid C, Isochlorogenic acid A, 1,3-Dicaffeoylquinic acid, etc.) in the alcohol extract were closely related with both anti-inflammatory and antioxidant activity, while the polysaccharides were also shown a certain anti-inflammatory and antioxidant activity. In conclusion, this study suggests that the flavonoids, phenolic acids and polysaccharides from chrysanthemum stem and leaf extracts can improve inflammatory bowel disease of zebrafish by regulating the expressions of IL-1β, IL-8 and MMP9. [ABSTRACT FROM AUTHOR]

Jujube (Ziziphus jujuba Mill.) fruit (JF) is widely consumed as food in Asian countries due to its potential effects for human health. As a traditional Chinese medicine, JF is often used to treat anorexia, fatigue and loose stools caused by spleen deficiency syndromes in China, but the mechanism underlying this effect has not been thoroughly elucidated. In this study, a rat model of spleen deficiency syndromes was adopted to investigate the therapeutic effect of JF extract and its possible mechanism by metabolomics analyses of plasma and urine as well as the intestinal flora analysis. The results showed that the changes in plasma and urine metabolites caused by spleen deficiency were reversed after administration of JF, and these changed endogenous metabolites were mainly involved in retinol metabolism, pentose and glucuronate interconversions, nicotinate and niacinamide metabolism pathways. The 16S rDNA sequencing results showed that JF could regulate intestinal flora imbalance caused by spleen deficiency. The covariance analysis of intestinal flora structure and metabolome indicated that Aerococcus may be a candidate strain for predicting and treating the metabolic pathways of spleen deficiency and related disorders. In summary, it can be revealed that spleen deficiency, which alters metabolic profiles and the intestinal flora, could be alleviated effectively by JF extract. [ABSTRACT FROM AUTHOR]

The seed of Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou (ZSS) is often used as a traditional Chinese medicine for insomnia due to its sedative and hypnotic effects, but the mechanism underlying this effect has not been thoroughly elucidated. In this study, an insomnia model induced by intraperitoneal injection of DL-4-chlorophenylalanine suspension in Sprague-Dawley rats was adopted to investigate the therapeutic effect of ZSS extract. Metabolomics analyses of plasma and urine as well as 16S rRNA gene sequencing of the intestinal flora were performed. The relationships between the plasma and urine metabolites and the intestinal flora in insomnia rats were also analyzed. The results showed that changes in plasma and urine metabolites caused by insomnia were reversed after administration of ZSS, and these changes were mainly related to amino acid metabolism, especially phenylalanine metabolism. The results of 16S rRNA gene sequencing and short-chain fatty acid determination showed that the ZSS extract could reverse the imbalance of intestinal flora caused by insomnia and increase the contents of SCFAs in feces. All of these improvements are mainly related to the regulation of inflammation. Therefore, it is concluded that insomnia, which alters metabolic profiles and the intestinal flora, could be alleviated effectively by ZSS extract. [ABSTRACT FROM AUTHOR]

Mulberry leaf was reported that it has antidiabetic activity, although the mechanisms underlying the function have not been fully elucidated. In the present study, the results of network pharmacology suggested that mulberry leaves could regulate key biological process in development of diabetes, and the process implicates multiple signaling pathways, such as JAK-STAT, MAPK, VEGF, PPAR, and Wnt. Then, the research in vitro indicated that mulberry leaves remarkably ameliorated high glucose-induced epithelial to mesenchymal transition, which was characterized with significant reduction of intracellular reactive oxygen species (ROS) levels as well as downregulation of NADPH oxidase subunits NOX1, NOX2, and NOX4, and it was found to be connected with the ERK1/2 signaling pathway in human tubular epithelial cells (HK-2). Moreover, the results of bioinformatics and the dual luciferase report showed that ZEB1 might be a target gene of miR-302a; decreased miR-302a and increased ZEB1 expressions could significantly promote epithelial to mesenchymal transition. However, mulberry leaves could reverse these modulations. Our results demonstrated that network pharmacology could provide a guidance role for traditional Chinese medicine research, and mulberry leaves could be of benefit in preventing high glucose-induced EMT in HK-2 cells, which proved that it was related to the upregulation of miR-302a by targeting ZEB1 and the inhibition of NADPH oxidase/ROS/ERK1/2 pathway. [ABSTRACT FROM AUTHOR]

Plant-microorganism interaction in the rhizosphere is thought to play an important role in the formation of soil fertility, transformation and absorption of nutrients, growth and development of medicinal plants, and accumulation of medicinal ingredients. Yet, the role that they play in the phthalides accumulation of Angelica sinensis (Oliv.) Diels remains unclear. In the present study, we report a correlative analysis between rhizosphere microorganisms and phthalides accumulation in A. sinensis from Gansu, China where was the major production areas. Meanwhile, Bacillus was explored the potential functions in the plant growth and phthalide accumulation. Results revealed that the common bacterial species detected in six samples comprised 1150 OTUs which were involved in 368 genera, and predominant taxa include Actinobacteria , Acidobacteria , and Proteobacteria. The average contents of the six phthalides were 4.0329 mg/g. The correlation analysis indicated that 20 high abundance strains showed positive or negative correlations with phthalides accumulation. Flavobacterium , Nitrospira , Gaiella , Bradyrhizobium , Mycobacterium , Bacillus , RB41 , Blastococcus , Nocardioides , and Solirubrobacter may be the key strains that affect phthalides accumulation on the genus level. By the plant-bacterial co-culture and fermentation, Bacillus which were isolated from rhizosphere soils can promote the plant growth, biomass accumulation and increased the contents of the butylidenephthalide (36∼415%) while the ligustilide (12∼67%) was decreased. Altogether, there is an interaction between rhizosphere microorganisms and phthalides accumulation in A. sinensis , Bacillus could promote butylidenephthalide accumulation while inhibiting ligustilide accumulation. [ABSTRACT FROM AUTHOR]

Although Scrophulariae Radix (root of Scrophularia ningpoensis) has received much attention, little is known about the nonmedicinal parts of S. ningpoensis. A comprehensive evaluation of the multibioactive constituents in the flowers, rhizomes, leaves, and stems of S. ningpoensis during different growth stages would be of value to fully understand the potential medicinal properties of all parts of the plant. Ultra‐high‐performance liquid chromatography coupled with triple quadrupole mass spectrometry was performed for accurately determining nine compounds in S. ningpoensis. The results indicated the content of total analytes in S. ningpoensis was in the order of flowers (81.82 mg/g) > roots (31.95 mg/g) > rhizomes (26.68 mg/g) > leaves (16.86 mg/g) > stems (14.35 mg/g). The chemometric analysis showed that these plant parts were rich in iridoids and should not be discarded during the processing of medicinal materials. Dynamic accumulation analysis suggested that the early flowering stage was the optimum time for harvesting flowers and appropriate amounts of stems and leaves. Moreover, considering the accumulation of constituents and biomass of medicinal materials, the medicinal parts should be harvested around December with the rhizomes attached. This research provides a theoretical basis and scientific evidence for comprehensive development and utilization of S. ningpoensis resources. [ABSTRACT FROM AUTHOR]

In this study, a rapid and highly sensitive ultra high performance liquid chromatography with triple quadrupole mass spectrometry method with the mobile phase of acetonitrile and 0.1% aqueous formic acid was established and successfully applied to comparatively analyze main active components after their compatibility. Besides, the effects of Scutellariae Radix, Coptidis Rhizoma and combined extracts on type 2 diabetic rats induced by high‐fat diet along with low dose of streptozocin were investigated. Under the optimized chromatographic conditions, good separation of seven target components was achieved within 12 min. All calibration curves exhibited good linearity (R2 ≥ 0.999). The relative standard deviation of precision, repeatability and stability varied from 0.69 to 2.23, 0.98 to 2.56, and 0.92 to 2.57%, respectively. The recovery ranged from 91.11 to 105.35%. The contents of seven active components were notably reduced after compatibility; however, the hypoglycemic effect of combined extracts was stronger than single drug by decreasing the activities of fructose‐1,6‐bisphosphatase, glucose 6‐phosphatase, phosphoenolpyruvate carboxykinase and increasing the activities of glucokinase, phosphofructokinase, pyruvate kinase. Accordingly, the established analytical method was accurate and sensitive enough for quantitative evaluation of seven investigated compounds. Moreover, the combined extract had definite effects on type 2 diabetes through multiple components against multiple targets. [ABSTRACT FROM AUTHOR]

A rapid and sensitive ultra high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry method was established and employed to determine 21 nucleosides, nucleobases, and amino acids in 60 samples from different parts of Angelicae Sinensis Radix. The established methods were validated by good linearity (r2 > 0.9937), limits of detection (0.12–77.75 ng/mL), limits of quantitation (0.31–272.13 ng/mL), intra‐ and interday precisions (RSD ≤ 4.84%, RSD ≤ 6.26%), stability (RSD ≤ 5.92%), repeatability (RSD ≤ 7.14%), recovery (91.4–103.4%), and matrix effects (0.92–1.03). Chemical comparative analysis revealed that the content of total analytes in four parts of Angelicae Sinensis Radix were different, and exhibited the order: Head (14.89 mg/g) > Body (10.15 mg/g) > All (8.22 mg/g) > Tail (6.23 mg/g). Principal component analysis showed that the samples could be classified into four groups in accord with four different parts of Angelicae Sinensis Radix. The results could provide a scientific basis and reference for the quality control of Angelicae Sinensis Radix, and may be conducive to further research on the pharmacological activities of Angelicae Sinensis Radix. [ABSTRACT FROM AUTHOR]

Rehmannia glutinosa Libosch (RG), is officially listed in the Chinese Pharmacopoeia and is widely used in China. In this paper, a sensitive and rapid ultra-performance liquid chromatography-mass spectrometry method including multiple-reaction monitoring mode was developed and applied to study the pharmacokinetic effect of acteoside from total glycoside extracted from the leaves of Rehmannia (TLR) and Dihuangye total glycoside capsule (DTG) in normal and diabetic nephropathy rats. The diabetic nephropathy rat model was induced by intraperitoneal injection of a small dose of streptozotocin and high-fat diet and plus 5% glucose drinking water. Samples of plasma of rats were obtained at different times after rats were administered TLR (7.2 g/kg) and DTG (360 mg/kg). After deproteinization by acetonitrile, the concentrations of acteoside in rats at different time points were detected by UPLC-TQ-MS method and pharmacokinetics parameters were calculated using DAS 3.2.8 software. A good linearity of acteoside was shown in the range of 8.51-3404.8 ng/m L ( r2 = 0.9987). The mean extraction recovery of analyte was in the range of 63.55-79.49%, and the intra- and inter-day RSD values were <8.8%. Compared with the normal group, the maximum plasma concentration, AUC0- t, AUC0-∞ and apparent plasma clearance corresponding dose in model group rats decreased significantly. After rats were administered TLR and DTG, the acteoside reached the maximum plasma concentration at about 15 min. The method proved to be simple, rapid and specific, and to be suitable for the determination of acteoside in plasma of diabetic nephropathy rats and pharmacokinetic study. [ABSTRACT FROM AUTHOR]

Flos Abelmoschus manihot is a traditional herbal medicine widely used in clinical practice to tackle chronic kidney disease (CKD) for thousands of years. Nowadays, many studies indicate that gut bacteria are closely related to the progression of CKD and CKD-related complications. In this study, a UPLC-Q-TOF/MS method coupled with the MetaboLynx™ software was established and successfully applied to investigate the metabolites and metabolic profile of Flos A. manihot extract by intestinal bacteria from normal and CKD rats. Eight parent components and eight metabolites were characterized by their protonated ions. Among these compounds, 15 were detected in the two group samples while M16 was only determined in the CKD model samples. Compared with the quercetin-type glycosides, fewer myricetin-type and gossypetin-type metabolites were obtained in the two group samples. These metabolites suggested that deglycosylation and methylation are the major metabolic pathways of Flos A. manihot extract. Few differences of metabolite classes were observed in the two group samples. However, the concentrations of aglycones such as quercetin, myricetin and gossypetin in the normal samples were notably higher than those in the CKD model samples. The results are important in unravelling the pharmacological effects of A. manihot and clarifying its mechanism of action in vivo. [ABSTRACT FROM AUTHOR]

Flos Chrysanthemi, dried flower of Chrysanthemum morifolium Ramat, has drawn much attention recently owing to its potential beneficial health effects for human. Flos Chrysanthemi products are usually taken orally and metabolized by intestinal microflora. However, there has been no investigation of the comprehensive metabolic profile of the Flos Chrysanthemi extract by intestinal flora owing to its chemical complexity and the limitations of analytical methods. In this paper, a rapid, sensitive and automated analysis method, ultra-performance liquid chromatography/quadrupole time of flight mass spectrometry including MSE technology and automated data processing Metabolynx™ software, was developed and successfully applied for the biotransformation and metabolic profile of flavonoids in the Flos Chrysanthemi extract by intestinal flora from human and rat. A total of 32 metabolites were detected and tentatively identified in human and rat intestinal bacterial samples. These metabolites indicated that hydrolysis, hydroxylation, acetylation, methylation, hydrogenation and deoxygenation were the major conversion pathways of flavonoids in the Flos Chrysanthemi extract in vitro. Furthermore, the effects of the Flos Chrysanthemi extract on the growth of different intestinal bacteria were detected using an Emax precision microplate reader. Certain pathogenic bacteria such as Enterobacter, Enterococcus, Clostridium and Bacteroides were significantly inhibited by Flos Chrysanthemi, while commensal probiotics such as Lactobacillus and Bifidobacterium were moderately promoted. Our observation provided further evidence for the importance of intestinal bacteria in the metabolism and potential activity of the Flos Chrysanthemi extract. The results will also be helpful for the further pharmacokinetic study of Flos Chrysanthemi and to unravel how it works in vivo. [ABSTRACT FROM AUTHOR]

Platycodin D (PD), a bioactive triterpenoid saponin isolated from Platycodi Radix (PR), possesses a vast range of biological activities. Although the pharmacological activities and pharmacokinetics of PD have been well demonstrated, information regarding the intestinal metabolisms of PD is very limited. In this study, human and rat fecal microflora were prepared and anaerobically incubated with PD at 37C for 48h, respectively. A highly sensitive and specific ultra performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was developed for the analysis of PD and related metabolites in the reaction samples. A Liquid-liquid extraction method was used for sample pretreatment and the chromatographic separation was performed on a 1.7 m particle size Syncronis C column using gradient elution system. Finally, a total of seven metabolites were detected and tentatively identified, such as the demethylation metabolite (M1), deoxidation metabolites (M3, M7) and hydrolysis at the C-28 oligosaccharide metabolites (M5, M6), which were first discovered in this experiment. The results indicate that hydrolysis, demethylation, dehydroxylation, and acetylation were the major metabolic pathways of PD in vitro. Additionally, four bacterial strains from human feces including Enterococcus sp.41, Bacillus sp.46, Escherichia sp.49 A and Escherichia sp.64 were detected and further identified with 16S rRNA gene sequencing due to their relatively strong metabolic capacity toward PD. The present study provides important information about the metabolism of PD, which will help elucidate the impact of intestinal bacteria on this active component. [ABSTRACT FROM AUTHOR]

The leaves of Ginkgo biloba L. have received much attention, whereas there has been little systematic analysis of the cortex and xylem from roots and branches. A comprehensive evaluation of the 44 compounds in the cortex and xylem would thus be of value to fully understand the potential medicinal properties of roots and branches. An assay of amino acids, terpene lactones, flavones, and phenolic acids was accomplished using ultra high performance liquid chromatography with tandem mass spectrometry. All of the calibration curves showed good linear regression ( R2 > 0.9902) within the tested ranges. The intra- and interday precision was less than 4.9% and the accuracy was within ±6.8%. The amount of terpene lactones in the cortex was 1.75-2.07-fold higher than that in the leaves. The amount of glutamine (360 μg/g) in the taproot xylem was 2.64-fold higher than that in the leaves (136 μg/g). Principal component analysis decreased in the order leaves > taproot cortex > rootlet > laterals cortex > branch cortex > stem cortex > taproot xylem > branch xylem > laterals xylem > stem xylem. The taproot of G. biloba might provide a supplementary source of terpene lactones, especially ginkgolide A and C, and of glutamine. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Traditional Chinese Medicines (TCMs) are increasingly used in combination with Western medicine. Scutellaria baicalensis Georgi (Lamiaceae) is a widely used TCM in treating various diseases. However, the in vivo metabolism of its main bioactive flavonoids, baicalin, baicalein, wogonoside and wogonin, needs further study. Materials and methods A systematic method based on ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technique combined with Metabolynx TM software was developed to speculate the metabolites and excretion profiles of the main flavonoids in S. baicalensis extract in rats bile, urine and feces samples after oral administration of the extract. Results Four parent components and a total of 15 metabolites were tentatively detected in vivo . All metabolites were detected including sulfate and glucuronide conjugates, hydroxylated, methylated, acetylated and deoxygenated products. Twelve metabolites were from the rat urine, five from the feces and two from the bile. Among them, several products were reported firstly. Conclusion The research provided useful information for further study of the pharmacology and mechanism of action of S. baicalensis extract in vivo and a proposed method which could develop an integrated template approach to analyze screening and identification of biological samples after oral administration of TCMs. [ABSTRACT FROM AUTHOR]

An ultra high performance liquid chromatography coupled with a triple quadrupole mass detection and electrospray ionization mass spectrometry method has been established for the simultaneous determination of the 14 nucleosides and nucleobases, 24 amino acids and two main alkaloids in mulberry leaf. In this method, a complicated mobile phase, the flow rate of which was 0.4 mL/min, was applied to the gradient elution, which provided a satisfied separation of the 40 compounds. The present method was validated, and sufficient reproducibility and accuracy were obtained for the quantitative measurement of the 40 compounds. The method was subsequently applied to ten mulberry leaves and the results showed that almost all of these samples were rich in nucleosides, nucleobases, amino acids, and alkaloids. The proposed method, which is convenient and economical, could serve as a prerequisite for the quality control of mulberry leaf herbs and be applied analogously to other Chinese medicines. [ABSTRACT FROM AUTHOR]

Context: Primary dysmenorrhea (PDM), a common, clinically heterogeneous endocrine disorder affecting young women, is associated with endocrinopathy and metabolic abnormalities. The Xiang-Fu-Si-Wu Decoction (XFSWD) is a traditional Chinese medicine preparation used to treat PDM. Objective: In the current study, a plasma metabonomics method based on the ultra-high-performance liquid chromatography-quantitative time-of-flight-mass spectrometry (UHPLC-Q-TOF-MS) system was employed to examine the mechanism of XFSWD action in PDM. Materials and methods: Estradiol benzoate (0.01 g/kg/d) and oxytocin (5 mL/kg) were used to create the dysmenorrhea rat model. Based on the chromatographic data of plasma samples at different time-points following oral administration of XFSWD mixed in water (37.8 g crude herbs/kg) on day 7, partial least square (PLS) and discriminate analysis (DA) were applied to visualize group differentiation and marker selection. Results: Systemic changes occurring in PDM reflect alterations in not only uterus function but also whole-body metabolism. The XFSWD was effective as a therapeutic agent for PDM by reflect metabolic pathway. Prostaglandins and lysophospholipids were identified as two marker types for oxytocin-induced dysmenorrhea syndrome, including LysoPC(18:4), LysoPE(22:2/0:0), LysoPC(17:0), PGJ2, 11-deoxy-11-methylene-PGD2, 15-deoxy-δ-12,14-PGJ2, LysoPC(20:3), etc. Specifically, the concentrations of prostaglandins compounds (PGJ2, 11-deoxy-11-methylene-PGD2, 15-deoxy-δ-12,14-PGJ2) were increased while those of lysophospholipid compounds [lysoPC(18:4), LysoPE(22:2/0:0), LysoPC(17:0)] were decreased to a significant extent ( p < 0.05) in dysmenorrheal rats. Upon treatment with the XFSWD at 12 h, the concentrations of lysophospholipids showed no significant differences ( P > 0.05) between the model and normal groups. The lysophospholipid levels were restored. Lysophospholipids were the key factors in phospholipid metabolism. Thus, disruption of phospholipids metabolism appears critical for the development of dysmenorrhea. The XFSWD exerted its effects by interfering with the sphingolipid metabolic pathway. Discussion and conclusions: The metabonomics method presents a promising tool to treat PDM in animal models, and may be applicable for clinical treatment of the human disease in the future. [ABSTRACT FROM AUTHOR]

A simple and specific UPLC-Q-TOF-MS method with automated data analysis (MetaboLynx) has been developed and applied for the primary investigation of metabolism pathway of active Xiang-Fu-Si-Wu Decoction fraction (XFSW-8). Through the online ESI-MSEanalysis and automated data analysis (MetaboLynx), a total of fifteen constituents have been identified or tentatively characterized from the nine tested samples after oral administration of XFSW-8. Six parent compounds and nine metabolites were identified in blood and eight organs. Methylates/demethylates sulfation, glucuronidation, sulfation, and glucuronidation conjugates were the major metabolic pathways of alkaloids in XFSW-8 in vivo. The present study provided important information regarding the metabolism of alkaloids in Xiang-Fu-Si-Wu Decoction which will be helpful for fully understanding of the mechanism of action of this Chinese medicine. Supplemental materials are available for this article. Go to the publisher's online edition ofthe Journal of Liquid Chromatography and Related Technologiesto view the free supplemental file. [ABSTRACT FROM AUTHOR]

ABSTRACT Introduction The pollen of Typha angustifolia L. has been used traditionally for the treatment of dysmenorrhea, stranguria and metrorrhagia in China. Recently, nucleosides and nucleobases have been proven as important bioactive compounds. Exploration of the nucleoside and nucleobase profiles from the pollen of T. angustifolia is important for improving its therapeutic value and could be convenient for its quality evaluation. Objective To establish an UPLC-PDA-MS method for simultaneous determination of nucleosides and nucleobases in the pollen of T. angustifolia. Methodology The analysis was performed on an Acuity UPLCHSS T3 column with a gradient elution of 5 m m ammonium acetate and methanol solution at a flow rate of 0.3 mL/min. Results Satisfactory separation of these compounds was obtained in less than 12 min. All calibration curves showed good linear regression ( r2 > 0.9995). The method provided good accuracy, precision, recovery, and sensitivity for the quantification of the 10 compounds analysed. Conclusion The UPLC method established is very helpful for optimising their content and could be convenient for quality evaluation of the pollen of T. angustifolia, which has not been reported as far as we are aware. Copyright © 2011 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

A validated liquid chromatography (LC) method coupled with hierarchical clustering analysis (HCA) has been established for investigating the fingerprint chromatograms of ten kinds of animal horns. The method showed good performances in terms of precision, repeatability, linearity, accuracy, and stability. The results showed that the LC fingerprint method coupled with HCA could be used to classify animal horns, which could make it simple and feasible to search substitution of precious horns. [ABSTRACT FROM PUBLISHER]