Ganoderma lucidum ethanol extracts ameliorate hepatic fibrosis and promote the communication between metabolites and gut microbiota g_Ruminococcus through the NF-κB and TGF-ß1/Smads pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Ganoderma lucidum, a traditional edible medicinal mushroom, has been widely reported to improve liver diseases as a dietary intervention for people. Ganoderma lucidum extracts, primarily total triterpenoids (GLTTs), are one of the bioactive ingredients that have excellent beneficial effects on hepatic fibrosis. Therefore, its prevention and reversal are particularly critical due to the increasing number of patients with chronic liver diseases worldwide. AIM OF THE STUDY: The study aimed to evaluate whether GLTTs had a hepatoprotective effect against hepatic fibrosis through metabolic perturbations and gut microbiota changes and its underlying mechanisms. MATERIALS AND METHODS: The compound compositions of GLTTs were quantified, and carbon tetrachloride (CCl4)-induced hepatic fibrosis rats were used to investigate the cause of the improvement in various physiological states with GLTTs treatment, and to determine whether its consequent effect was associated with endogenous metabolites and gut microbiota using UPLC-Q-TOF-MSE metabolomics and 16S rRNA gene sequencing technology. RESULTS: GLTTs alleviated physical status, reduced liver pathological indicators, proinflammatory cytokines, and deposition of hepatic collagen fibers via regulating the NF-κB and TGF-ß1/Smads pathways. The untargeted metabolomics analysis identified 16 potential metabolites that may be the most relevant metabolites for gut microbiota dysbiosis and the therapeutic effects of GLTTs in hepatic fibrosis. Besides, although GLTTs did not significantly affect the α-diversity indexes, significant changes were observed in the composition of microflora structure. In addition, Spearman analysis revealed strong correlations between endogenous metabolites and gut microbiota g_Ruminococcus with hepatic fibrosis. CONCLUSION: GLTTs could provide a potential target for the practical design and application of novel functional food ingredients or drugs in the therapy of hepatic fibrosis.

An UPLC - Q - Orbitrap method for pharmacokinetics and tissue distribution of four triterpenoids in rats after oral administration of Poria cocos ethanol extracts.

Poria cocos (Schw.) Wolf, is a fungus that is widely used as medicine and dietary supplement in China. But its action mechanism is still not very clear. In this paper, a rapid, specific and sensitive high performace liquid chromatography coupled with hybrid quadrupole - orbitrap mass sepctrometry (UPLC - Q - Orbitrap MS) method has been developed and validated to simultaneously determine of four triterpenoids including Dehydrotumulosic acid (DTA), Dehydropachymic acid (DPA), Pachymic acid (PA), Dehydrotrametenolic acid (DMA) from Poria cocos in rat plasma and tissues. The analyte was extracted from rat plasma and tissue homogenates by protein precipitation with acetonitrile using glibenclamide as the internal standard (IS). Chromatographic separation was carried out on ACQUITY UPLC BEH - C18 column (2.1 mm × 50 mm, 1.7 µm) with a mobile phase composed of acetonitrile - water (containing 1.0 mmol/L ammonium acetate) using gradient elution at a flow rate of 0.2 mL/min. Electrospray ionization (ESI -) under negative ion mode was used, and its quantization was performed with multiple reaction monitoring (MRM) mode. The method was fully validated and successfully applied to pharmacokinetics and tissue distribution study in rats after oral administration of ethanol extracts of Poria cocos. Compared with that of plasma exporsure, triterpenoids could be detected in various tissues with a relatively high degree of tissue distribution. After oral administration, the concentration orders in seven different tissues were ranked as DTA > PA > DPA > DMA in intestine and stomach, wheras DTA > DMA > PA > DPA in heart, liver, spleen, lung and kidney tissues, which is speculated that DPA, PA may be converted into DMA in vivo. In conclusion, this results may provide a material basis for study of the pharmacological actions of triterpenoids in Poria cocos.

Simultaneous determination of the carboxylate and lactone forms of 10-hydroxycamptothecin in human serum by restricted-access media high-performance liquid chromatography.

A simple restricted-access media (RAM) HPLC method for simultaneous determination of the lactone and carboxylate forms of 10-hydroxycamptothecin (HCPT) in human serum was established. Using a RAM Hisep analytical column, serum samples were directly injected into the HPLC system. The eluted peaks of two forms of HCPT were monitored with a fluorescence detector. The separation was completed in 17 min. The linear range was 20-1000 ng/ml, intra-day and inter-day variations being less than 5%. The kinetic equation was introduced according to the analytical results. The equation shows that the course of the HCPT lactone form converting to carboxylate form in human serum at 4 degrees C is a first-order kinetic course. The concentration of each form at the moment of sampling was calculated by extrapolation.

Restricted-access media high pressure liquid chromatography vs fluorescence polarization immunoassay for analysis of carbamazepine in human plasma.

AIM: To compare restricted-access media high performance liquid chromatographic (RAM-HPLC) method with fluorescence polarization immunoassay (FPIA) for analysis of carbamazepine (CBZ) in human blood. METHODS: An RAM-HPLC method was established for the determination of CBZ in plasma. RESULTS: The two methods do not need sample clean-up prior to analysis and they have almost 100 % recovery and good reproducibility. There is a good correlation between the CBZ concentration in venous plasma samples determined by FPIA and that in both venous and fingertip plasma samples obtained by RAM-HPLC, the correlation coefficients being 0.989 and 0.995, respectively. It is shown by t-test that the data sets of venous and fingertip plasma samples given by RAM-HPLC are consistent with each other but significantly different from the results obtained by FPIA. CONCLUSION: Both direct injection RAM-HPLC and FPIA may be applied in determining CBZ in therapeutic drug monitoring (TDM). FPIA is well-suited to the routine TDM. RAM-HPLC is more useful in TDM related research and especial cases.