•An UPLC-MS/MS method was developed for simultaneous quantitation of 8 saikosaponins.•The method was successfully applied to pharmacokinetic study in depression rat.•PK on two saikosaponins from Bupleuri Radix was first carried out.•Comparison between PK of raw and vinegar-baked Bupleuri Radix was first studied.•The result showed that vinegar-baked could change absorption of some saikosaponins.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 0.40 mL/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 0.62 ng/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.

BackgroundAlthough flowers of Panax ginseng Meyer (FPG), Panax quinquefolius L. (FPQ), and Panax notoginseng Burk. (FPN) have been historically used as both medicine and food, each is used differently in practice.MethodsTo investigate the connection between components and enhancing immunity activity of FPG, FPQ, and FPN, a method based on a rapid LC coupled with quadrupole time-of-flight MS and immunomodulatory activity study evaluated by a carbon clearance test were combined.ResultsAccording to quantitative results, the ratio of the total content of protopanaxatiol-type ginsenosides to protopanaxadiol-type ginsenosides in FPN was 0, but ranged from 1.10 to 1.32 and from 0.23 to 0.35 in FPG and FPQ, respectively. The ratio of the total content of neutral ginsenosides to the corresponding malonyl-ginsenosides in FPN (5.52 ± 1.33%) was higher than FPG (3.2 ± 0.64%) and FPQ (2.39 ± 0.57%). The colorimetric analysis showed the content of total ginsenosides in FPQ, FPG, and FPN to be 13.75 ± 0.60%, 17.45 ± 0.42%, and 12.45 ± 1.77%, respectively. The carbon clearance assay indicated that the phagocytic activity of FPG and FPQ was higher than that of FPN. A clear discrimination among FPG, FPQ, and FPN was observed in the principal component analysis score plots. Seven compounds were confirmed to contribute strongly by loading plots, which may be the cause of differences in efficacy.ConclusionThis study provides basic information about the chemical and bioactive comparison of FPG, FPQ, and FPN, indicating that protopanaxtriol-type ginsenosides and malonyl-ginsenosides may play a key role in their enhancing immunity properties.

•Influence of drying on phenolics contents and antioxidants was studied.•19 analytes were determined to investigate the influence of drying.•The optimized drying method for Cimicifuga dahurica was oven-drying at 110 °C.•The effective constituents contributing to antioxidant activity were ensured.Cimicifuga dahurica (Turcz.) Maxim is usually used as traditional Chinese medicine and functional tea. In present work, the change of its phenolics profiles and antioxidant activities during drying processes were investigated. Nineteen main bioactive analytes of C. dahurica were simultaneously quantified by HPLC-DAD to assess the effect of drying method. Meanwhile, the influence of different drying processes on antioxidant activities were evaluated by DPPH (1,1-diphenyl-2-picrylhydrazyl), ABTS [2,2′-Azino-bis-(3-ethylbenzthiazoline-6-sulphonate)], FRAP (ferric reducing antioxidant power) and hydroxyl radical scavenging method. Results showed that drying method had significant effects on the contents of phenolics and antioxidant properties. The appropriate drying method was oven-dried at 110 °C. Moreover, the content of phenolic acids and antioxidant activity showed significant correlation coefficient. And the most effective constituents contributing to antioxidant activity were isoferulic acid (8), cimicifugic acid B (14) and 2-isoferuloyl piscidic acid (16).

Three new compounds (1, 6, 9), with six known compounds (2–5, 7–8) were obtained from water-soluble extract of Cimicifuga dahurica (Turcz.) Maxim. by bioactivity-guided isolation. Their structures were elucidated by chemical and spectral analysis, including 1D, 2D NMR data and HRESIMS. H2O2-induced neurotoxicity on PC12 cells model were conducted to evaluate the neuro-protective capability of these compounds. The piscidic acid derivatives compounds 4–7 showed marked neuro-protective effect at certain concentration.Download high-res image (109KB)Download full-size image

Bupleuri Radix (BR) is a traditional Chinese medicine (TCM) widely used in Asian nations, which originates from Bupleurum chinense DC or Bupleurum scorzonerifolium Willd. It can be baked with vinegar to obtain vinegar-baked Bupleuri Radix (VBBR). It has been reported that VBBR exhibits more potential activity for relieving the effects of a depressed liver than BR. However, the antidepressant effects and underlying pharmacological mechanisms of BR and VBBR remain unclear. The present study identified the main chemical compositions in BR and VBBR. Furthermore, behavioral studies along with biochemical assessments, as well as 1H NMR analysis of the hippocampus and liver were employed to systematically assess the pathology of depression and therapeutic effects of BR and VBBR using a rat model of chronic unpredictable mild stress (CUMS)-induced depression. Behavioral studies demonstrated that treatment with VBBR resulted in better antidepressant effects than BR. And VBBR significantly regulated the levels of neurotransmitters in the hippocampus and frontal cortex. Additionally, twelve and ten metabolites from the hippocampus and liver were identified as potential biomarkers associated with depression. Such biomarkers are primarily involved in energy metabolism, amino acid metabolism, glycolysis, inositol phosphate metabolism, lipid metabolism and the TCA cycle. The results presented here showed that VBBR significantly reversed the pathological process of CUMS-induced depression, partially via regulation of the disturbed metabolic pathways. The comparative study facilitated a better understanding of the antidepressant mechanisms of BR and VBBR.

•The Ononin inhibited the productions of NO, PGE2, TNF-α, IL-1β and IL-6.•The Ononin inhibited mRNA expression of COX-2 and iNOS.•The Ononin inhibited the phosphorylation of IκBα and MAPKs.Increasing evidence has shown that ononin, a major isoflavone, has anti-inflammatory effects on lipopolysaccharide (LPS)-induced inflammation. However, the molecular mechanisms underlying the anti-inflammatory effects of ononin are still unclear. In the present study, we investigated these effects and the underlying mechanisms of ononin on LPS-induced inflammatory responses. Mouse RAW 264.7 cells were treated with 1 μg/mL LPS and 5, 25, 50, 100 or 150 μM ononin for 18 h. Cell viability was assessed using MTT assays, and the production of nitric oxide (NO), prostaglandin E2 (PGE2) and the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 in cultures was examined by Griess and ELISA analyses. qRT-PCR was performed to detect the mRNA expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2). Mitogen-activated protein kinases (MAPKs) and nuclear transcription factor Kappa-B (NF-κB) signalling pathway-related proteins were assessed by western blot assays. The results showed that cell viability was not significantly affected by up to 100 μM ononin. The production of NO, PGE2 and the pro-inflammatory cytokines TNF-α, IL-1β and IL-6 in the cultures, the mRNA expression of two major inflammatory mediators, COX-2 and iNOS, and the expression of phosphorylated IκB-α, ERK, JNK, and p38 MAPKs proteins in LPS-treated cells were significantly increased. These changes could be reversed by treatment with ononin in a concentration-dependent manner (P < 0.05). The results suggest that ononin has anti-inflammatory effects on LPS-induced inflammatory responses by inhibiting the NF-κB and MAPK pathways and may be a potential treatment for inflammation.Download high-res image (145KB)Download full-size image

Three new phenolic compounds (1–3), along with five known compounds (4–8) were isolated from the rhizome of Cimicifuga dahurica (Turcz.) Maxim. Their structures were elucidated by spectroscopic methods including 1D-NMR, 2D-NMR and HR-MS techniques. DPPH method and protective effect on PC12 cells against H2O2-induced oxidative damage model were carried to evaluate the antioxidant capability of these compounds. Compound 5 showed significant antioxidant activity with IC50 values 9.33 μM in DPPH assay and compound 2 displayed marked neuro-protective effect with 87.65% cell viability at the concentration of 10 μM. Additionally, the possible structure-activity relationships of these phenolic compounds were tentatively discussed.Download high-res image (300KB)Download full-size image

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 (5 mmol/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 10−6 cm/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.