Study On The Extraction Process Of Jingning Granules And The Intestinal Absorption Of Four Lignans In The Formulation

Objective1.According to determination of Verbascoside,Tenuifolin,Schizandrol A,Schizandrol B,Schisandrin A and Schisandrin B contents,we selected the best extraction process.2.The Everted gut sac method combined with HPLC-DAD detection was used to determine the main components of JingNing particle absorbed into the intestine.3.The Oil-Water partition coefficient,Equilibrium solubility and intestinal absorption dynamic of the four kinds of lignans in JingNing particle,including Schizandrol A,Schizandrol B,Schisandrin A and Schisandrin B were measured.Methods1.Prepare the five kinds of samples and determine the concents of Verbascoside,Tenuifolin,Schizandrol A,Schizandrol B,Schisandrin A and Schisandrin B.2.The Everted gut sac technique combined with HPLC-DAD detector was employed to determine the main components of JingNing particle absorbed into the intestine.3.The log P of Schisandrol A,Schisandrol B,Schizandrin A and Schizandrin B dissolved in pure water and a series of phosphate buffered saline solutions with increasing pH values(pH 1.0,2.0,2.5,5.0,5.8,6.5,7.8 and 9.0)were determined using the shake-flask technique.4.In situ intestinal recirculating perfusion model of JingNing particle was performed to study its absorption with different concentrations.Calculate the Ka,T1/2,P and Papp of the four lignans.5.JingNing particle in different concentrations were perfused in situ in each intestinal segment of rats.Effective permeability(Papp)of Schisandrol A,Schisandrol B,Schizandrin A and Schizandrin B in each segment was calculated and the site of absorption was determined.JingNing particle with different pH was also perfused in suit into the whole intestine of rats.In situ single-passed intestinal perfusion was used to study compatible effects of Schisandra chinensis and herb-pairs(polygala tenuifolia extracted by alcohol,volatile oil of Rhizoma Acori Tatarinowii and other drugs extracted by water)on intestinal absorption of Schizandrol A,Schizandrol B,Schisandrin A and Schisandrin B in the low concentration.Results1.A specific HPLC method was developed for the simultaneous analysis of Verbascoside,Tenuifolin,Schisandriol A,Schisandrol B,Schizandrin A and Schizandrin B.The retention time were 6.651,22.754,39.057,44.175,67.763,74.690 min.The recovery rate was between 95%and 105%.The RSD of precision,repeatability stability were less than 3%.The values of craft one～five were 0.7154,2.1154,2.1778,4.3828,4.4481 mg/g.2.Result shows that a large part of components come from Schisandra chinensis were absorbed into intestine.3.The log P of Schisandrin A dissolved in pure aqua and a sequence of phosphate buffered saline solutions with increasing pH values(pH 1.0,2.0,2.5,5.0,5.8,6.5,7.8 and 9.0)were 2.19,2.22,2.34,2.72,2.35,2.29,2.30,2.33,2.26,Schisandrin B were 3.30,3.01,3.16,3.17,3.11,3.03,3.12,3.13,3.08,Schizandrin A were 2.26,2.06,1.95,1.88,2.24,2.32,1.68,2.06,2.53,Schizandrin B were 2.05,2.31,2.01,2.37,2.01,2.42,2.31,2.22,2.48.4.The intestinal recirculating perfusion model showed that Ka has no significant difference.5,The in situ single-passed intestinal perfusion showed,the sequence of Ka is low>middle>high.The absorption tends to be decreased in the order of ileum,duodenum and jejunum.The best pH is 6.5.Results showed that polygala tenuifolia could significantly enhance the absorption of Schizandrol A,Schizandrol B(2.5 times),Schisandrin A(3.8 times)and Schisandrin B(2.8 times).Volatile oil of Rhizoma Acori Tatarinowii could enhance the absorption of Schizandrol A(3.0 times),Schizandrol B(1.9 times),Schisandrin A(3.4 times)and Schisandrin B(2.4 times).But there had no significant difference for the water extraction of other drugs.Conclusion1.The best craft is Schisandra chinensis and polygala tenuifolia are extracted by 60%Alcohol,rhizoma acori graminei naphtha is needed,and the rest of the medicine are extracted by water.2.Both the intestinal recirculating perfusion and in situ single-passed intestinal perfusion showed that there maybe a carrier intermediary transport way.The four lignans could be well absorbed in general intestinal tract,particularly the ileum.3.Polygala tenuifolia could promote the absorption of the four lignans distinctly,followed by rhizoma acori graminei.