Characterization and discrimination of raw and vinegar-baked Bupleuri radix based on UHPLC-Q-TOF-MS coupled with multivariate statistical analysis.

Bupleuri Radix is a commonly used herb in clinic, and raw and vinegar-baked Bupleuri Radix are both documented in the Pharmacopoeia of People's Republic of China. According to the theories of traditional Chinese medicine, Bupleuri Radix possesses different therapeutic effects before and after processing. However, the chemical mechanism of this processing is still unknown. In this study, ultra-high-performance liquid chromatography with quadruple time-of-flight mass spectrometry coupled with multivariate statistical analysis including principal component analysis and orthogonal partial least square-discriminant analysis was developed to holistically compare the difference between raw and vinegar-baked Bupleuri Radix for the first time. As a result, 50 peaks in raw and processed Bupleuri Radix were detected, respectively, and a total of 49 peak chemical compounds were identified. Saikosaponin a, saikosaponin d, saikosaponin b3 , saikosaponin e, saikosaponin c, saikosaponin b2 , saikosaponin b1 , 4''-O-acetyl-saikosaponin d, hyperoside and 3',4'-dimethoxy quercetin were explored as potential markers of raw and vinegar-baked Bupleuri Radix. This study has been successfully applied for global analysis of raw and vinegar-processed samples. Furthermore, the underlying hepatoprotective mechanism of Bupleuri Radix was predicted, which was related to the changes of chemical profiling.

Validated UPLC-MS/MS method for simultaneous quantification of eight saikosaponins in rat plasma: Application to a comparative pharmacokinetic study in depression rats after oral administration of extracts of raw and vinegar-baked Bupleuri Radix.

A sensitive and efficient UPLC-MS/MS method was developed and validated for simultaneous quantification of eight saikosaponins (SSa, SSb1, SSb2, SSb3, SSb4, SSc, SSd and SSf) in rat plasma. Furthermore, comparative pharmacokinetic profiles of these sakosaponins were investigated, following oral administration of extracts of raw and vinegar-baked Bupleuri Radix to depression rats. Biosamples were processed with liquid-liquid extraction technique using ethyl acetate. Chromatographic separation was accomplished on Waters BEH C18 UPLC column. 0.05% formic acid in water and acetonitrile as mobile phase was used at a flow rate of 0.40mL/min. The analytes and internal standard, digoxin, were detected using negative ion electrospray ionization in multiple reaction monitoring mode. The lower limit of quantification was less than 0.62ng/mL for the each analyte. The validation parameters investigated, which were specificity, precision, accuracy, matrix effect, recovery and stability, were well within acceptable limits. Results showed that for some of analytes, AUC0-t and Cmax were significantly different after oral administration of extracts of raw and vinegar-baked Bupleuri Radix. Moreover, the pharmacokinetic study in pathological state could provide more useful information to guide the clinical usage of Bupleuri Radix.

Pharmacokinetic studies of active triterpenoid saponins and the total secondary saponin from Anemone raddeana Regel.

The rhizome of Anemone raddeana Regel, a Traditional Chinese Medicine (TCM) which has a robust history treating rheumatism and neuralgia. The total secondary saponin (TSS) from it has demonstrated antitumor activity. In this study, a rapid and validated LC-MS/MS method was developed to simultaneously determine the active compounds (Hederacolchiside A1 and Eleutheroside K). Analytes were separated on a reverse-phase C18 column with acetonitrile-water (5mmol/L ammonium acetate) as the mobile phase. This assay showed acceptable linearity (r>0.99) over the concentration range 5-1000 nmol/L for two analytes. The intra- and inter-day precision was within 8.06% and accuracy was ranged from -3.16% to 3.34% for two analytes. The mean extraction recoveries of analytes and IS from rat plasma were all more than 76.0%. Under the developed analytical conditions, the obtained values of main pharmacokinetic parameters (Cmax and AUC0-t) indicated that the pure compounds were more efficient than the TSS extract in Hederacolchiside A1 and Eleutheroside K absorption. In addition, pharmacokinetic studies of two individual compounds demonstrated their poor oral absorption in rat (aF%, 0.019-1.521). In the study of absorption and transportation of Hederacolchiside A1 and Eleutheroside K in Caco-2 cell monolayer model, the uptake permeability was in 10-6cm/sec range suggesting poor absorption, which confirmed the previous pharmacokinetic profiles in vivo. Interestingly, the uptake ratio of them declined significantly when treated with phloridzin (SGLT1 inhibitor). It indicated that the absorption of Hederacolchiside A1 in intestine was mainly through positive transport and SGLT1 might participate in its active absorption.

Chemical constituents from rhizome of Anemone amurensis.

Phytochemical investigation of the 70% EtOH extract of the rhizome of Anemone amurensis led to the isolation of two new oleanane-type triterpenoid saponins 1 and 2. Their structures were elucidated on the basis of chemical and spectral analysis, including 1D, 2D NMR data, and HR-ESI-MS. Compounds 1 and 2 were tested for cytotoxicities against two human cancer cell lines (A549 and Hep-G2). Compound 2 showed potent cytotoxicity with IC50 values of 38.53 and 66.17 µM, respectively, while compound 1 with IC50 > 100 µM.

New flavonoid glycosides from Sedum aizoon L.

Five new flavonoid glucosides (3-4, 10-12) and a new phenolic derivative (5), together with eight known compounds including three flavonoid glucosides (6-8), three phenolic compounds (1-2, 9) and two megastigmane glucosides (13, 14), were isolated from the ethanol extract of the aerial part of Sedum aizoon L. Among them, compounds 9, 13 and 14 were isolated and identified from this genus for the first time. The structures of compounds were elucidated on the basis of 1D and 2D NMR (HSQC, HMBC and COSY) spectra and the HR-ESI-MS data. These compounds were tested for their antibacterial efficacies against both Gram-positive and Gram-negative bacteria. Compounds 1, 2, 3, 7 and 10 showed certain antibacterial activity; it showed more potency against Gram-positive than against Gram-negative bacteria. Compound 2 showed the most pronounced antibacterial effectiveness against Staphylococcus aureus Rosenbach with MIC value of 7.8µg·mL(-1). The in vitro anti-proliferative activities against HepG2, MCF-7 and A549 tumor cell lines were also evaluated. The result suggested compound 7 exhibited moderate cytotoxic activities with IC50 values of 46.30, 75.27 and 49.76µmol/L, respectively.

Two new triterpenoid saponins from rhizome of Anemone amurensis.

Two new triterpenoid saponins were isolated from the 70% ethanol extract of the rhizome of Anemone amurensis, they are oleanolic acid 28-O-ß-d-glucopyranosyl-(1 → 3)-α-l-rhamnopyranosyl-(1 → 4)-ß-d-glucopyranosyl-(1 → 6)-ß-d-glucopyranosyl ester (1) and 23,27-dihydroxy oleanolic acid 3-O-α-l-arabinopyranoside (2). The structures of 1 and 2 were elucidated on the basis of chemical and spectral analysis, including 1D and 2D NMR data and HR-ESI-MS. Compounds 1 and 2 were tested for cytotoxicities against three human cancer cell lines (A549, Hep-G2, and MCF-7). Compound 1 showed potent cytotoxicity with IC50 values of 34.76, 41.17, and 28.92 µM, respectively, while compound 2 with IC50>100 µM.