Screening of quality markers and activity verification of Glycyrrhizae Radix et Rhizoma based on small molecule compound-protein interaction / 中国中药杂志

In order to solve the problem of weak correlation between quality control components and efficacy of Glycyrrhizae Radix et Rhizoma, this study detected the interaction between small molecular chemical components of Glycyrrhizae Radix et Rhizoma and total proteins of various organs of mice by fluorescence quenching method to screen potential active components. The 27 chemical components in Glycyrrhizae Radix et Rhizoma were detected by HPLC and their deletion rates in 34 batches of Glycyrrhizae Radix et Rhizoma were calculated. Combined with the principle of component effectiveness and measurability, the potential quality markers(Q-markers) of Glycyrrhizae Radix et Rhizoma were screened. RAW264.7 macrophage injury model was induced by microplastics. The cell viability and nitric oxide content were detected by CCK-8 and Griess methods. The levels of inflammatory factors(TNF-α, IL-1β, IL-6, CRP) and oxidative stress markers(SOD, MDA, GSH) were detected by the ELISA method to verify the activity of Q-markers. It was found that the interaction strength between different chemical components and organ proteins in Glycyrrhizae Radix et Rhizoma was different, reflecting different organ selectivity and 18 active components were screened out. Combined with the signal-to-noise ratio of the HPLC chromatographic peaks and between-run stability of the components, seven chemical components such as liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin, liquiritigenin, isoliquiritigenin and ammonium glycyrrhizinate were finally screened as potential Q-markers of Glycyrrhizae Radix et Rhizoma. In vitro experiments showed that Q-markers of Glycyrrhizae Radix et Rhizoma could dose-dependently alleviate RAW264.7 cell damage induced by microplastics, inhibit the secretion of inflammatory factors, and reduce oxidative stress. Under the same total dose, the combination of various chemical components could synergistically enhance anti-inflammatory and antioxidant effects compared with the single use. This study identified Q-markers related to the anti-inflammatory and antioxidant effects of Glycyrrhizae Radix et Rhizoma, which can provide a reference for improving the quality control standards of Glycyrrhizae Radix et Rhizoma.

Establishment and verification of method for detection of hydrogen sulfide in organs by fluorescent probe method / 中国药理学通报

Aim To rapidly prepare and purify hydrogen sulfide specific fluorescent probe (WSP-5), establish and optimize the fluorescent probe method for the determination of hydrogen sulfide in animal tissues, and verify the applicability of the method in the model of malignant pleural effusion. Methods The preparation solvent of fluorescent probe reaction solution, DMSO addition volume, pH, reaction solution solvent and reaction solution volume, sample pretreatment temperature, grinding times, and standing time after grinding were investigated. The mouses model of malignant pleural effusion was established with S-180 ascites tumor cells, and the concentration of hydrogen sulfide in various organs and tissues of the model animal was measured. Results After optimization, silica gel and dextran gel were used as stationary phases, dichloromethane methanol formic acid (60: 1: 0.1, V/V/V) and dichloromethane methanol (1: 1, V/V) were used as eluents for separation and purification, and the first eluting component was taken to prepare WSP-5 with a purity of more than 700 mg. Animal tissue samples and sodium hydrosulfide standard solution were added with 5 times of cold reaction solution, after low temperature vibration grinding, highspeed centrifugation, the supernatant was incubated in dark for 12 hours, the fluorescence intensity was measured by fluorescent microplate reader. Hydrogen sulfide concentration was calculated according to the standard curve. The LOD of this method was about 0. 6 |JLmol • L

Determination of nine kinds of nucleosides in Spirodelae Herba from different habitats by UPLC-MS/MS / 中草药

Objective: To establish a UPLC-MS/MS method for the simultaneous determination of nine kinds of nucleosides in Spirodelae Herba, and analyze and investigate the difference of nucleosides ingredients in samples from 13 habitats. Methods: The determination was performed on the Waters XbridgeTM Amide-C18 (150 mm × 4.6 mm, 3.5 μm), the 0.1% formic acid-water (A) and 0.1% acetonitrile-water (B) in gradient elution as mobile phase. The flow rate was 0.5 mL/min. The column temperature was 30 ℃. MS instrument was equipped with ESI+ ion source. Gaining the extracted ion chromatograms, then the peak area for quantitative, principal component analysis (PCA), and hierarchical clustering analysis (HCA) were used to comprehensive evaluation. Results: All nine kinds of nucleoside showed good linearity (r ≥ 0.998 9). The RSD of the precision, repeatability, and stability tests were less than 3.5%. The average recovery rates were in the range of 94.4%-101.9%, RSD were in the range of 1.73% - 3.58%. There are differences in the composition and content from different habitats. The content of 2'-deoxyuridine and 2'-deoxyinosine in the lower levels, cytosine was the lowest, while the contents of uridine, inosine, and xanthine were in higher levels. The largest content was in Spirodelae Herba from Huangshan, Anhui province. Conclusion: The method for the simultaneous determination of the nine kinds of nucleosides is simple, accurate, and reproducible, that will provide the basis for the formulation of herbs Spirodelae Herba for quality control standards.

Multi-level identification and analysis about infrared spectroscopy of lophatheri herba / 中国中药杂志

Based on the infrared spectra of Lophatheri Herba and Commelinae Herba, one-dimensional infrared spectra, second derivative spectra and two-dimensional correlated spectra were used to find out the differences between Lophatheri Herba and its imitations, respectively. The common peak ratio and variant peak ratio dual-indexes sequential were calculated and established according to infrared spectra of eleven batches of herbs. Infrared spectral data of Lophatheri Herba cluster analysis was applied to explore the similarity between each sample. The grouping results trend of sequential analysis of dual-indexes and cluster analysis was accordant. The results showed that the differences could be found by multi-level identification, and the source and the quality of the herbs could be effectively distinguished by the two analysis methods. Infrared spectroscopy, used in the present work exhibited some advantages on quick procedures, less sample required, and reliable results, which could provide a new method for the identification of traditional Chinese medicine with the imitations and adulterants, and the control of quality and origin.