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1.
Article in Chinese | WPRIM | ID: wpr-906467

ABSTRACT

Objective:The differences of chemical compositions and pharmacological activities between the core and pulp of Phyllanthi Fructus were investigated by chemical analysis and <italic>in vitro</italic> test to explore the effect of the core on the quality of this medicinal material. Method:Literature, medicinal material standards and market research on the appearance of Phyllanthi Fructus were conducted based on existing databases. Ultra-high performance liquid chromatography-quadrupole-electrostatic field orbital trap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) was used to identify the constituents of the core and pulp. The analysis was performed on Thermo Scientific Accucore C<sub>18</sub> column (2.1 mm×100 mm, 2.6 μm) with the mobile phase of 0.1% formic acid aqueous solution (A)-methanol (B) for gradient elution (0-25 min, 5%B; 25-30 min, 5%-95%B; 30-35 min, 95%-5%B), the flow rate was 0.2 mL·min<sup>-1</sup>, heating electrospray ionization (HESI) was adopted with positive and negative ion modes, and the scanning range was <italic>m</italic>/<italic>z</italic> 100-1 500. High performance liquid chromatography (HPLC) was used to determine the contents of gallic acid, corilagin, chebulagic acid and ellagic acid in the core and pulp of Phyllanthi Fructus. Analysis was performed on Welchrom C<sub>18</sub> column (4.6 mm×250 mm, 5 μm) with mobile phase of methanol (A)-0.05% phosphoric acid aqueous solution (B) for gradient elution (0-6 min, 5%A; 6-15 min, 5%-7%A; 15-20 min, 7%-15%A; 20-25 min, 15%-21%A; 25-31 min, 21%-22%A; 31-41 min, 22%A; 41-47 min, 22%-28%A; 47-51 min, 28%-32%A; 51-57 min, 32%-38%A; 57-70 min, 38%-45%A; 70-80 min, 45%-65%A; 80-85 min, 65%-5%A), the detection wavelength was set at 270 nm. The antibacterial effects of the core and pulp of Phyllanthi Fructus on <italic>Escherichia coli</italic> and <italic>Staphylococcus aureus</italic> were investigated by filter paper method, and their antioxidant activities were compared by 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging assay. Result:A total of 47 compounds were identified in the core and pulp of Phyllanthi Fructus, mainly including tannins, flavonoids, phenolic acids, fatty acids, amino acids, organic acids, saccharides and glycosides, most of which were concentrated in the pulp, and the fatty acids in the core accounted for a higher proportion. The contents of gallic acid, corilagin, chebulagic acid, ellagic acid and other phenolic compounds in the pulp of 20 batches of Phyllanthi Fructus were much higher than those in the core. The results of antibacterial test showed that the core of Phyllanthi Fructus with different concentrations had no antimicrobial effect. The DPPH radical scavenging test showed that the antioxidant activity of the core [half-inhibitory concentration (IC<sub>50</sub>)=199.632 mg·L<sup>-1</sup>] was much less than that of the pulp (IC<sub>50</sub>=12.688 mg·L<sup>-1</sup>). Conclusion:From the perspectives of polyphenol content, antibacterial and antioxidant activities, it is scientific to use Phyllanthi Fructus pulp in ancient and modern times, which may be to remove the secondary parts of Phyllanthi Fructus, so as to enhance the actual utilization rate and therapeutic effect of medicinal materials. In view of the large proportion of the core of Phyllanthi Fructus and its high content of fatty acids and other components, whether or not to use it remains to be further studied in clinical application.

2.
Article in Chinese | WPRIM | ID: wpr-906438

ABSTRACT

Objective:To provide a scientific basis for the classification of Phyllanthi Fructus product grades. Method:A total of 30 batches of Phyllanthi Fructus currently available in the market were collected for quantification based on such appearance indexes as diameter, thickness, grain weight, and crust colour (<italic>L</italic><sup>*</sup>, <italic>a</italic><sup>*</sup>, and <italic>b</italic><sup>*</sup> values). The contents of gallic acid, corilagin, chebulagic acid, and ellagic acid were measured by high performance liquid chromatography (HPLC), followed by descriptive statistical analysis (DSA), analysis of variance (ANOVA), and principal component analysis (PCA) to determine the importance of each main index and explore the correlations between the appearance indexes and internal components. The classification standard of Phyllanthi Fructus product grades was formulated, and its scientificity was verified in hepatocelular carcinoma HepG2 cells. Result:The correlation analysis revealed that the crust colour <italic>L</italic><sup>*</sup>, <italic>a</italic><sup>*</sup>, and <italic>b</italic><sup>*</sup> values were significantly negatively correlated with corilagin, chebulagic acid, and ellagic acid (<italic>|r|</italic>>0.5, <italic>P</italic><0.01), but irrelevant to gallic acid (<italic>|r|</italic><0.1). Considering the variable coefficient of each index, PCA results, and the requirement of gallic acid as quality indicator for Phyllanthi Fructus in <italic>Chinese Pharmacopoeia</italic>, the crust colour <italic>L</italic><sup>*</sup>, <italic>a</italic><sup>*</sup>, and <italic>b</italic><sup>*</sup> values and gallic acid content were determined to be the classification indexes. The K-means cluster analysis confirmed that products with crust colour <italic>L</italic><sup>*</sup><44, <italic>a</italic><sup>*</sup><7, and <italic>b</italic><sup>*</sup><10 and gallic acid content >1.6% could be classified into the first class, and those failing to meet the above requirements into the second class. The cell experiment demonstrated that the half-maximal inhibitory concentration (IC<sub>50</sub>) of the first-class product against hepatocelular carcinoma HepG2 cells was lower than that of the second-class product. A colourimetric card was developed based on crust colour <italic>L</italic><sup>*</sup>, <italic>a</italic><sup>*</sup>, and <italic>b</italic><sup>*</sup> values to provide a visual tool for on-site evaluation of Phyllanthi Fructus products. Conclusion:This study has initially established the classification standard of Phyllanthi Fructus product grades, which contributes to guiding price negotiation of Phyllanthi Fructus products based on quality grade and thus ensuring high quality and high price.

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