Liquid chromatography-mass spectrometry-based strategy for systematic profiling of chemical components and associated quantitative analysis of quality markers in Qi-Wei-Tong-Bi oral liquid.

Qi-Wei-Tong-Bi oral liquid (QWTB), a famous Chinese medicine preparation composed of seven crude drugs has a good therapeutic effect on rheumatoid arthritis and is widely used in China. However, its chemical composition and quality control have not been comprehensively and systematically investigated. In this study, high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was employed for its chemical profiling. As a result, 100 components were chemically characterized. Additionally, high-performance liquid chromatography coupled with a quadrupole linear ion trap mass spectrometry method was developed to simultaneously quantify nine bioactive components (hyperoside, ononin, quercetin, sinomenine, magnoflorine, gallic acid, protocatechuic acid, monotropein, and cyclo-(Pro-Tyr)) in multiple-reaction monitoring mode. After successful validation in terms of linearity, precision, repeatability, and recovery, the assay method was applied for the determination of 10 batches of QWTB. The results showed that QWTB was enriched in sinomenine and magnoflorine with the highest amount up to hundreds or even thousands of µg/mL, while quercetin, ononin, cyclo-(Pro-Tyr), and hyperoside were much lower with the lowest content below 10 µg/mL. This study work would help to reveal the chemical profiling and provide a valuable and reliable approach for quality evaluation and even pharmacodynamic material basis studies of QWTB.

Comparative analysis of the rhizosphere microbiome and medicinally active ingredients of Atractylodes lancea from different geographical origins.

This study aimed to explore the important role of the rhizosphere microbiome in the quality of Atractylodes lancea (Thunb.) DC. (A. lancea). The rhizosphere microbial community of A. lancea at two sampling sites was studied using metagenomic technology. The results of α-diversity analysis showed that the rhizosphere microbial richness and diversity were higher in the Maoshan area. The higher abundance of core microorganisms of the rhizosphere, especially Penicillium and Streptomyces, in the Maoshan area compared with those in the Yingshan area might be an important factor affecting the yield of A. lancea. Redundancy analysis illustrated that the available phosphorus had a significant effect on the rhizosphere microbial community structure of A. lancea. We also showed that the plant-microbe and microbe-microbe interactions were closer in the Maoshan area than in the Yingshan area, and Streptomyces were the main contributors to the potential functional difference between the two regions. A. lancea in the Maoshan area had a high content of atractylodin and atractylon, which might be related to the enhanced abundance of Streptomyces, Candidatus-Solibacter, and Frankia. Taken together, this study provided theoretical insights into the interaction between medicinal plants and the rhizosphere microbiome and provides a valuable reference for studying beneficial microbes of A. lancea.

Extraction, structural characterization, and biological activities of a new glucan from Codonopsis pilosula.

In this study, a powerful and rapid aqueous two-phase system (ATPS) method was used to extract polysaccharides from Codonopsis pilosula. The ATPS process was investigated with response surface methodology (RSM). At an ammonium sulfate concentration of 17%, ethanol concentration of 30%, and extraction temperature of 40 °C at pH 6, the total extraction yield of polysaccharides reached (31.57 ± 1.28)%. After separation and purification, a homogenized polysaccharide CPP 2-4 with molecular weight of 3.9 × 104 kDa was obtained from the bottom phase. The physicochemical properties and structural features confirmed that CPP 2-4 was an α-1,6-glucan. Activity studies showed that the IC50 of CPP 2-4 for DPPH radical scavenging was 0.105 mg/mL. The FRAP and ABTS assays showed that CPP 2-4 had strong antioxidant activity in a dose-dependent manner. Furthermore, CPP 2-4 inhibited NO release in RAW264.7 cells induced by lipopolysaccharide, which indicated a certain anti-inflammatory effect. This study improved the extraction rate of polysaccharides from C. pilosula and identified a glucan for the first time, that can contribute to a better understanding of the composition and structure of polysaccharides from C. pilosula and provide data support for the medicine and food homology of C. pilosula.

Mechanism exploration of ancient pharmaceutic processing (Paozhi) improving the gastroprotective efficacy of Aucklandiae Radix.

ETHNOPHARMACOLOGICAL RELEVANCE: Processing, also called Paozhi in Chinese, is an ancient Chinese pharmaceutic processing technique developed along with the Chinese herbal medicines (CHMs). The understanding of the mechanism of Paozhi has been investigated for several decades. Aucklandiae Radix (CAR) and its roasted processed products are all used in indigestion as a kind of CHMs. Processed Aucklandiae Radix (PAR) had a stronger effect to protect gastric mucosa than CAR, while the main compounds in CAR were reduced sharply after being processed. The underlying mechanism of this phenomenon is still unclear. AIM OF THE STUDY: This study was aimed to evaluate whether PAR have a stronger gastroprotective effect than CAR and the underlying mechanisms of such circumstance. MATERIALS AND METHODS: Ultra-fast liquid chromatography coupled with quadrupole time of flight mass spectrometry (UFLC-QTOF-MS/MS) coupled with multivariate statistical analyses was employed to explore chemical compounds which had a relatively stable content in PAR. Based on the compounds selected as the research object, network pharmacology was applied to visualize the relationships between the selected components and the gastroprotective-related targets from disease database, at the same time the possible intervention path of CAR/PAR which might be responsible for the effect of CAR/PAR on gastritis-induced rats was also built. Then, the key proteins were detected by western blotting to verify and compare the pharmacological effects of CAR/PAR. RESULTS: Through UFLC-QTOF-MS/MS and orthogonal partial least squares discriminant analysis (OPLS-DA), sixteen compounds stable in PAR were discovered, of which saussureamine C and saussureamine B were estimated as the core compounds to exert gastroprotective in PAR predicted by network pharmacology analysis. Under the guide of KEGG pathway enrichment analysis, PI3K/AKT, p38 MAPK (Mitogen-activated protein kinase) and nuclear factor-kappa B (NF-κB) signaling pathways were forecasted as the possible healing mechanisms of CAR/PAR, and that result was verified by the experiments in vivo. PAR performed a stronger ability to reduce the level of p38 MAPK and NF-κB p65 than CAR, which may partially explain the different ability of CAR/PAR against gastric mucosa damage. CONCLUSION: This study clarified that although Paozhi entailed a sharp decrease on the main compounds of CAR, there were some compounds which were not sensitive to high temperature and preserved in PAR and had a relative higher content in PAR than in CAR. PAR has stronger influence on MAPKs/NF-κB signaling pathway than CAR, which may reveal that the stronger gastroprotective effect of PAR perhaps rely on the constitutions with a higher relative abundance after Paozhi. The present research combined UFLC-QTOF-MS/MS and network pharmacology deeply investigated the impact of the roasted processing on the chemical constitutions and gastroprotective effect of CAR and offered reference for the clinical application of CAR/PAR.

Metabolomics analysis of dandelions from different geographical regions in China.

INTRODUCTION: Dandelion (Taraxacum mongolicum Hand.-Mazz.) is a perennial herb with diverse pharmacological effects. The development and utilization of dandelion have attracted much attention. OBJECTIVES: Our aims were to provide a reference basis for the identification of the origin of dandelions and to study the influence of their origin on their quality. Methods High-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was used to analyze metabolites from dandelions from four different geographical regions in China, namely Gansu, Henan, Shanxi, and Jiangsu. Metabolite analysis was performed using orthogonal partial least-squares discriminant analysis, and to identify potential metabolic pathways, MBRole was used to perform Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. RESULTS: Principal component analysis revealed that the chemical components of dandelions sampled from the four regions showed noticeable differences. Twenty-six, six, six, eight, eight, and fifteen differentially produced metabolites were identified upon comparison between Gansu and Jiangsu, Gansu and Shanxi, Gansu and Henan, Henan and Shanxi, Henan and Jiangsu, and Shanxi and Jiangsu, respectively. These differentially produced metabolites were mainly phenolic compounds. Further, KEGG pathway enrichment analysis showed that the main metabolic pathways involved were biosynthesis of phenylpropanoids and flavonoids. CONCLUSION: The methods reported herein can be used to identify the origin of dandelions; moreover, our results can serve as a reference basis for future studies.

Resveratrol suppresses human cervical carcinoma cell proliferation and elevates apoptosis via the mitochondrial and p53 signaling pathways.

Numerous studies have demonstrated the apoptotic and anti-proliferative effects of resveratrol, a natural polyphenolic phytoalexin, on various cancer cell lines. However, the effects of resveratrol on the regulation of human cervical carcinoma, and the mechanisms underlying these effects, remain to be elucidated. In the present study, the potential mechanisms underlying the effects of resveratrol in HeLa cervical carcinoma cells were investigated. The results revealed that resveratrol inhibited proliferation and induced apoptosis in HeLa human cervical cancer cells in a dose-dependent and time-dependent manner. Resveratrol induced cell shrinkage in HeLa cells and apoptosis accompanied by the activation of caspase-3 and -9. Furthermore, resveratrol upregulated the expression of the pro-apoptotic B-cell lymphoma (Bcl)-2-associated X protein and downregulated the expression of the anti-apoptotic proteins Bcl-2 and Bcl-extra large in HeLa cells. In addition, p53, a protein that is essential for cell survival and cell cycle progression, exhibited elevated expression levels in resveratrol-treated HeLa cells. Therefore, resveratrol may be a promising novel inhibitor of human cervical cancer.

[Macroscopic and Microscopic Identification of Ganoderma and The Confusable Medicinal Mushrooms of "Zhi"].

OBJECTIVE: To compare the macroscopic and microscopic characteristics of Ganoderma and the confusable medicinal mushrooms of "Zhi". METHODS: The methods of macroscopic and microscopic examination were employed to authenticate Ganoderma and the other medicinal mushrooms of "Zhi" in the fungal group. RESULTS: The medicinal mushroom named "Zhi" could be distinguished from each other by the shape of pileus, stipe and the characteristics of mycelium and spore. CONCLUSION: There are abundant resources of fungal group from Ganodermataceae in China. The medicinal mushrooms of "Zhi", which are quite complicated in the market, are needed to be differentiated.

[Study on HPLC-DAD Fingerprint of Xiaoying Jiedu Decoction].

OBJECTIVE: Based on backtracking method, identification and quality evaluation of Xiaoying Jiedu Decoction (XYJDD) by HPLC fingerprint analysis were carried out. METHODS: HPLC-DAD fingerprint of XYJDD was conducted with Dikma Platisil ODS C18 column (250 mm x 4.6 mm, 5 µm) and gradient elution with the mobile phase consisting of acetonitrile-(0.1% phosphoric acid-0.1% triethylamine solution) at column temperature of 30 °C. On the basis of the established chromatographic pattern of XYJDD tracking backward to the corresponding crude herbal drugs in the formula and the attribution of most peaks in the XYJDD fingerprint can be figured out. RESULTS: 16 peaks of XYJDD HPLC fingerprint were assigned by parallel comparison with the fingerprint of the five corresponding crude drugs in the formula of Scutellariae Radix, Glycyrrhizae Radix et Rhizoma, Houttuyniae Herba, Aurantii Fructus, Arnebiae Radix. Four peaks can be identified with the chemical reference substances. CONCLUSION: The entirety of XYJDD HPLC fingerprint enhances the specialty for control and assessment of the product quality, and the backtracking experimental method can be expected to be applied in chromatographic fingerprinting analysis of complex Chinese patent medicine.

[Chemical constituents of Juncus setchuensis].

OBJECTIVE: To study the chemical constituents of Juncus setchuensis. METHODS: Column chromatography was used in the isolation procedure. The structures of isolated compounds were elucidated by spectral data. RESULTS: Eight compounds were isolated and their structures were identified as 2-hydroxy-3-methylanthraquinone (1), physcion (2), stigmasterol (3), stigmast-3,6-dione (4), vanillin (5), n-heptacosanoic acid (6), trans-hydroxycinnamic (7) and 4-hydroxy-3-methoxy benzoic acid (8). CONCLUSION: Compound 1, 2, 4 and 6 are obtained from this genus for the first time and all the compounds are obtained from this plant for the first time.

[Study on the chemical constituents of Psoralea corylifolia].

OBJECTIVE: To study the chemical constituents of Psoralea corylifolia. METHODS: Column chromatography was used in the isolation procedure. The structures of isolated compounds were elucidated by spectral data. RESULTS: Seven compounds were isolated and their structures were identified as isopsoralen (1), psoralen (2), bavachalcone (3), 4", 5"-dehydroisopsoralidin (4), methyl 4-hydroxybenzoate (5), psoralidin (6), corylin (7). CONCLUSION: Compounds 4 and 5 are obtained from Psoralea for the first time.

[Determination of baicalin in Xiaoyanling Decoction by high-performance liquid chromatography].

OBJECTIVE: To establish a high-performance liquid chromatography (HPLC) method for the determination of baicalin in Xiaoyanling Decoction. METHODS: HPLC method was used for determination. A Kromosil C18 column was used. The mobile phase consisted of MeOH-acid water (H(2)O:H(3)PO(4)=53:0.2) 40:60, and the detective wavelength was 277 nm. RESULTS: The content of baicalin in each Xiaoyanling Decoction sample was not less than 0.40 mg/ml (RSD<5%). CONCLUSION: The content of baicalin in Xiaoyanling Decoction is high and stable, and it can be used as the quality standard for Xiaoyanling Decoction.

[Effect of ultra-fine powder technique on dissolution rates of andrographolide and dehydroandrographolide in Andrographis paniculata].

OBJECTIVE: To investigate the effects of ultra-fine powder technique on dissolution rates of the components in Andrographis paniculata. METHODS: High performance liquid chromatography was employed to determine the concentration of andrographolide and dehydroandrographolide in common powdered or ultra-fine powdered Andrographis paniculata. RESULTS: The dissolution rates of andrographolide and dehydroandrographolide in ultra-fine powdered Andrographis paniculata were higher than those of the general powder. CONCLUSION: Ultra-fine powder technique promotes the dissolution rates of andrographolide and dehydroandrographolide.