[Simultaneous determination of content of eight components in Caesalpinia decapetala by UPLC-MS/MS].

The present study established the ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) method for simultaneous determination of the content of eight major active components in Caesalpinia decapetala and performed the quality evaluation of C. decapetala from different habitats with the chemical pattern recognition. The analysis was carried out on a Waters BEH C_(18) column(2.1 mm×100 mm, 1.7 µm) at 40 ℃, with the mobile phase of water containing 0.1% formic acid(A) and acetonitrile containing 0.1% formic acid under gradient elution, the flow rate of 0.3 mL·min~(-1), and the injection volume of 1 µL. The electrospray ionization(ESI) source in the negative mode and multiple reaction monitoring(MRM) were used for MS quantitative analysis. The content results were analyzed by the hierarchical cluster analysis(HCA) and principal component analysis(PCA) for the evaluation of the quality difference. Eight components showed good linear relationships within their respective concentration ranges(r>0.999), with the average recoveries of 96.85%-103.4% and RSD of 0.52%-2.8%. The analysis results showed that the quality of samples from different batches was different. The samples were classified into three clusters by HCA and PCA. The method is simple, sensitive, accurate, and efficient, and can be used for the quality evaluation of C. decapetala.

[Pharmacokinetics of three index components of Periploca forrestii in rats by microdialysis combined with UPLC-MS/MS].

The present study established a UPLC-MS/MS method for the content determination of Periploca forrestii microdialysis samples and investigated the pharmacokinetics of three index components of P. forrestii in rats. The effects of flow rate and concentration of perfusate on the recovery rate were investigated by the concentration difference method(increment method and decrement method). The microdialysis samples at different time points were collected, and the concentrations of three index components were determined by UPLC-MS/MS. The actual drug concentrations were corrected with the in vivo recovery rate, and the pharmacokinetic parameters were calculated by WinNonlin 8.2. In the in vitro recovery test, the recovery rate measured by the increment method and the decrement method was inversely proportional to the flow rate and independent of the concentration. The pharmacokinetic parameter AUC_(0-t) values of 3-O-caffeoyl quinic acid, 4-O-caffeoyl quinic acid, and 5-O-caffeoyl quinic acid were(23 911.23±5 679.67),(16 688.43±3 448.45), and(9 677.02±1 606.74) min·µg·L~(-1), respectively. C_(max) values were(170.66±58.02),(121.61±48.14), and(69.69±18.23) µg·L~(-1), respectively. The UPLC-MS/MS method has the advantages of specificity, rapidity, high sensitivity, and accurate quantification. It can simultaneously determine the concentration of 3-O-caffeoyl quinic acid and other two index components in microdialysis samples and is suitable for the pharmacokinetics study of the three index components of P. forrestii in normal rats.

Biancaea decapetala (Roth) O.Deg. extract exerts an anti-inflammatory effect by regulating the TNF/Akt/NF-κB pathway.

BACKGROUND: Biancaea decapetala (Roth) O.Deg. (Fabaceae) is used to treat colds, fever, and rheumatic pain caused by inflammation. However, the mechanism underlying its anti-inflammatory properties remains unclear. PURPOSE: This study aimed to evaluate the anti-inflammatory activity of Biancaea decapetala extract (BDE) in vitro and in vivo and explore the possible underlying mechanism and potential targets. METHODS: The release of nitric oxide (NO) and inflammatory cytokines in LPS-stimulated RAW264.7 cells and rats were measured using Griess reagent and enzyme-linked immunosorbent assay (ELISA). Hematoxylin and eosin (H&E) staining was employed to examine the pathology of animal tissues. Transcriptome analysis was performed to screen the pathways related to BDE-mediated inhibition of inflammation, and the expression of related proteins was measured using real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, ELISA, and immunofluorescence methods. Surface Plasmon Resonance (SPR) and the Drug Affinity Reaction Target Stability (DARTS) method were used to verify whether BDE binds to TNF-α target protein, while a L929 cell model and NF-κB gene reporter systematic method were used to investigate the inhibitory effect of BDE on the activity of TNF-α protein. RESULTS: BDE inhibited the expression of TNF-α, IL-1ß, IL-6, and NO in RAW264.7 cells and rats, and improved the pathological changes in lung tissue. RNA-seq showed that BDE may regulate the TNF/Akt/NF-κB pathway to inhibit inflammation onset. BDE significantly downregulated the mRNA expression of TNF-α, IL-6, IL-1ß, and that of relevant proteins, including TNF-α, p-p65, p-Akt, p-IκBα. Furthermore, BDE inhibited the nuclear translocation of NF-κB (p65) and the activation of the Akt pathway by SC79. The L929 cell model, luciferase reporter gene analysis, DARTS, and SPR experiments showed that BDE may bind to TNF-α and inhibit the TNF-α-NF-κB pathway. CONCLUSION: BDE may target TNF-α to inhibit the TNF/Akt/NF-κB pathway, thereby attenuating inflammation. These findings reveal the anti-inflammatory effects and mechanisms of BDE and provide a theoretical basis for the further development and utilization of BDE.