Pharmacokinetic study on five components in phthalide target areas of Chaxiong and its β-CD inclusion compounds based on UPLC-MS/MS / 中国中药杂志

This study aims to establish a method for the determination of the concentration of five main components of phthalide target areas of Chaxiong(CPTA) and its inclusion of β-CD in the plasma of rats, and determine the pharmacokinetic parameters, absolute bioavailability and relative bioavailability of CPTA/β-CD inclusion compound in vivo. The plasma concentrations of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide were determined with UPLC-MS/MS. The content determination was conducted at the chromatographic conditions as follows: Shim-pack GIST C_(18)-AQ HP column(2.1 mm×100 mm, 3 μm), mobile phase of 0.1% formic acid solution(A)-acetonitrile(B), gradient elution, flow rate of 0.3 mL·min~(-1), column temperature of 35 ℃ and injection volume of 2 μL. The mass spectra were obtained with electrospray ion source(ESI), positive ion mode and multi reaction monitoring. CPTA/β-CD inclusion compound was prepared by grinding method, DAS 2.0 software was used to model the data, and the absolute bioavailability of CPTA and relative bioavailability of inclusion compound were calculated. Finally, the methods for the determination of five components of senkyunolide A, N-butylphthalide, new osthol lactone, Z-ligustilide and butenyl phthalide in CPTA, were successfully established. The linear relationship among the five components was good within their respective ranges, r>0.99. The absolute bioavailability of the five components in rats was 22.30%, 16.32%, 21.90%, 10.16% and 12.43%, respectively. After CPTA/β-CD inclusion was prepared, the relative bioavailability of the five components was 138.69%, 198.39%, 218.01%, 224.54% and 363.55%, respectively, significantly improved. This method is rapid, accurate and sensitive, so it is suitable for the pharmacokinetic study of extracts in traditional Chinese medicine and their preparations.

Chemical comparison on different parts of Angelica sinensis Radix based on NMR metabolomics / 中草药

Objective: To compare the chemical constituents in different parts of Angelica Sinensis Radix (ASR). Methods: In this study, NMR-based metabolomics approach was used to compare the chemical constituents of different parts in ASR. And the metabolites of different parts in ASR were analyzed using PCA, OPLS-DA, and univariate analysis. Results: Totally 36 metabolites were identified in the NMR spectra. Multivariate analysis revealed that higher levels of some compounds, such as valine and isoleucine, were present at the head of ASR, while the tail showed higher contents of Z-butylidenephthalide, cis-Z,Z'-3a.7a,7a.3a'- dihydroxyligustilide, etc. The body showed higher contents of tryptophan, coniferyl ferulate, and the levels of aspartic acid, Z-ligustilide, etc were present at higher levels in both head and tail. For some metabolites, such as alanine and arginine, no significant difference was observed among the three parts of ASR. Conclusion: Both the primary and secondary metabolites of ASR can be detected at the same time by 1H-NMR based metabolic profiling approach used in this study, and the results revealed in this study laid the foundation for the further investigation of the relationship between the chemical components and biological effects of different parts in ASR.

Protection of Z-ligustilide on human umbilical vein endothelial cells and its mechanism / 中草药

Objective: To observe the effect of Z-ligustilide on the expression of tumor necrosis factor-α (TNF-α)-induced human umbilical vein endothelial cells (HUVEC) intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and nuclear factor kappa B (NF-κB), and to investigate the protection on endothelial cells injured by inflammatory cytokines and its mechanism. Methods: TNF-α (10 ng/mL) was added to in vitro cultured HUVEC to induce the cell injury. After co-incubated with the different concentration (5, 10, and 20 μmol/L) of Z-ligustilide for 24 h, the cell viability in each group was measured by MTT. The levels of ICAM-1 and VCAM-1 were assayed using enzyme linked immunosorbent assay (ELISA). Western blotting was used to detect the expression of NF-κB protein of cell in each group. Results: TNF-α could induce the obvious injury of HUVEC compared with the control group (P < 0.05, 0.01), and increase the expression of ICAM-1 and VCAM-1 in HUVEC (P < 0.01). Compared with the model group, Z-ligustilide could dose-dependently reduce the expression of ICAM-1 (P < 0.01) and VCAM-1 (P < 0.05) and obviously decrease the NF-κB expression (P < 0.05). Conclusion: Z-ligustilide could attenuate TNF-α-induced injury in HUVEC via suppressing the activation of NF-κB and inhibiting the excessive expression of ICAM-1 and VCAM-1.

One step separation and preparation of senkyunolide A and Z-ligustilide in Ligusticum chuanxiong Hort by high speed counter current chromatography / 中成药

AIM: To research how to separate the active component in Ligusticum chuanxiong Hort by high speed counter current chromatography. METHODS: Senkyunolide A and Z-ligustilide,the main components of volatile oil in Ligusticum chuanxiong Hort were one step separated by high speed counter current chromatography. n-hexane-ethyl acetate-ethanol-water,1 ∶ 1 ∶ 1 ∶ 1(v/v),was used as the solvent system for HSCCC. Top phase and bottom phase were respectively used as static phase and mobile phase. Optimum speed and flow rate were 900 r/min and 1. 2 mL/min respectively. RESULTS: Collected fractions were analyzed by HPLC and identified by EI-MS and 1HNMR. Purity could reach more than 95% . CONCLUSION: Lactone is fit to be separated and prepared by high speed counter current chromatography with good resolution and high purity. We find a fast and efficient way to separate these.