Sensitive Analysis and Pharmacokinetic Study of Berberrubine Using LC-MS/MS

Objective To develop a sensitive and reproducible liquid chromatography-tandem mass spectrometry（LC-MS/MS） method to evaluate the pharmacokinetic behavior of berberrubine（BRB） and its glucuronide（BRBG） in rats. Methods BRB, BRBG and tetrahydroberberine（THB, internal standard） were isolated by liquid-liquid extraction in rat biological samples. Chromatographic separation was achieved on an Agilent Zorbax Eclipse Plus C_（18）（2.1 mm × 50 mm, 3.5-Micron） with a gradient mobile phases primarily containing acetonitrile, water with 0.1% formic acid and 5 mm ammonium acetate. The analytes were monitored by MS/MS in positive electrospray ionization mode. Herein, the feasibility of new developed method was validated with respect to specificity, linearity, precision, accuracy, stability, extraction efficiency and matrix effect. The appropriate method was used for the pharmacokinetic study in rats.Results The new developed method could be applied to the pharmacokinetic study of BRB in rats. BRB and BRBG showed good linearity over the ranges of 2-1000 ng/mL and 5-2000 ng/mL, respectively, and precision was no more than 15%. The accuracy, specificity and stability could be acceptable. Conclusion The new method is sensitive and reproducible. In pharmacokinetic study, BRB showed nonlinear elimination property. Meanwhile, BRB was rapidly absorbed and widely distributed in various tissues with the highest exposure of BRB in kidney and liver. The absolute bioavailability of BRB was determined to be 8.2% and at the dose of 40 mg/kg, a total of 44% BRB was excreted in urine and feces.

System pharmacology-based dissection of the potential effective material basis and mechanism of Xiaoxianxiong decoction for type 2 diabetes mellitus

Background:Xiaoxianxiong decoction is a classic formula in traditional Chinese medicine used to treat type 2 diabetes mellitus and proven to be effective.But the material basis and underlying mechanisms remain unclear.The aim of the present study was to elucidate the potential effective material basis of Xiaoxianxiong decoction and molecular mechanism treating type 2 diabetes mellitus.Methods:The absorbed bioactive components were identified based on serum pharmacochemistry.Network analysis were performed to obtain effect targets for docking verification with the absorbed prototypes to determine the potential effective material basis.On the above basis,network pharmacology was conducted to explore the molecular mechanism.Results:76 compounds were identified of Xiaoxianxiong decoction and 61 absorbed bioactive compounds were investigated.Serine/threonine kinase 1 and ALB were key targets acquired by network analysis for molecular docking.Subsequently,5 compounds were considered as the potential effective material basis,namely berberine,berberrubine,lariciresinol and gingerenone A,jatrorrhizine.Further,the mechanism mainly lies in the insulin signaling pathway,HIF-1 signaling pathway,PI3K-Akt signaling pathway,FoxO signaling pathway,AGE-RAGE signaling pathway in diabetic complications,phospholipase D signaling pathway to regulate blood glucose levels on target tissues as well as organs and exhibit anti-inflammatory,promote cell differentiation and cell growth,maintain oxygen homeostasis and affect the enzymes along with key metabolites.Conclusion:This integrated research strategy to investigate the treatment of Xiaoxianxiong decoction on type 2 diabetes mellitus provides valuable insights for further study and clinical practice of Xiaoxianxiong decoction.