Pharmacokinetics And Relative Bioavilablity Of Diammonium Glycyrrhizinate Capsules

The Hepatitis B (HB) caused by Hepatitis B virus (HBV) is a serious infectiousdisease. Diammonium Glycyrrhizinate has outstanding liver protecting and enzymereducing action. It aslo has anti-inflammatory and anti-fibrosis effects which lead toprevent liver cancer. Pharmacokinetics and relative bioavilablity of DiammoniumGlycyrrhizinate in Chinese Healthy Volunteers will be reported in this thesis, and itwill provide academic foundation for the development and clinical use ofDiammonium Glycyrrhizinate.Objective To sduty pharmacokinetics and relative bioavilablity of DiammoniumGlycyrrhizinate in Chinese Healthy Volunteers.Methods A 2-way crossover design was performed based on High-PerformanceLiquid Chromatography (HPLC). Twenty healthy male volunteers took 300 mgDiammonium Glycyrrhizinate capsules (the test product and the reference product) insingle doseing open label, randomized crossover sduty. The interval was one week.The character of volunteers included normal function of liver, renal, and heart, nohistory hypersceptibility. The volunteers were forbidden using other drugs, startingfrom 2 weeks before the test to the end of the test. After overnight fast, each of themtook a single 300mg oral dose diammonium glycyrrhizinate capsules with 200mLtepid water. A standerized meal of low fat and protien was served speratedly at 4hrs after the drug ingestion. Venous blood samples(5mL) were collected , immediatelybefore the drug administration and at 2,3,4, 6, 8,10,12,14,16, 24,36,48 hours afterdosing. The sammples were taken with heparin anticoagulant immediately after thecollection, and stored at -30℃until assayed. There were 18 volunteers drug plasma inthis report because the No.2 and No.20 didn't finish the process completely. Drugplasma concentration of glycyrrhetinic acid was examed by high-performance liquidchromatography (HPLC) assay. The sample processing: The heparised blood plasma2mL was taken accurately, 5ml ethyl acetate was added for extraction. Let it mixvortically for 3min, centrifuged at 4000rpm for 15min. The 4mL organic up-layer wasextracted accurately blowed dry with nitrogen around the 50℃water, then disslovedwith 100μL the mobile phase. And 50μL was sampled. The chromatographiccondition: Scienhome C₁₈ column(5μm, 4.6×250 mm) with the mobile phase ofacetonitrile: 10mmol·L^-1 ammonium acetate (55 : 45)(v/v) at a flow rate of1.0mL·min^-1, the UV detection wavelength was 254nm; with the temperature of 40℃.The plasma concentration-time data were disposed with BECS program(vol.1.0.2). The peak connection (C_max) and the peak time (T_max) of glycyrrhetinic acidwere determined from the respective observed concentration-time data. The area ofcurve (AUC) were calculated by the liner trapezoidal method. Diference inpharmacokinetics data between standard reference formulation and test formulationwere evaluated statistically by use of two-sample t test.Results The results showed that this method was specific and selective forglycyrrhetinic acid in plasma and impurity subtance in plasma didn't interfere withdetermination. Obtained linear regression equation of metformin is C =0.001078A-0.5717 (r = 0.9998) within the line range of 2.5～500μg·L^-1 forglycyrrhetinic acid in plasma. The relative recoveries, difference intra-day andintra-day all meet with the examination of living creature sample.The main pharmacokinetic parameters of the test and the reference formulation arefollowed: T_max were(13.67±4.77)h and (13.00±3.84) h, C_max were(116.30±61.78)μg·L^-1 and(118.64±89.01)μg·L^-1, t_1/2β were (13.56+10.99)h and (12.01±4.50) h, AUC₀^t were (1931.1±831.0)μg·h·L^-1 and (1959.5±904.8)μg·h·L^-1, AUC₀^∞were (2272.0+1202.0)μg·h·L^-1 and (2207.0+1093.0)μg·h·L^-1。relative bioavilablityof F₀^t was(102.1±25.51)%,and F₀^∞was(99.94±23.60)%. The results of Analysisof Variance (ANOVA) and two one-sided t test statistical analysis show thatdiammonium glycyrrhizinate capsules (the test product and the reference product) arebioequivalent.Conclusion1. The HPLC assay is shown to be sensitive, accurate and simple and the results arereliable.2. Diammonium Glycyrrhizinate capsules (the test product and the reference product)are bioequivalent.