Studies On The Pharmacokinetics And Quality Control Methods Of Polygonum Cuspidatum

Polygonum cuspidatum was studied in this dissertation.RP-HPLC method was developed for the simultaneous determination of polydatin,resveratrol,anthraglycoside B,emodin and physcion in Polygonum cuspidatum.The linear ranges for polydatin, resveratrol,anthraglycoside B,emodin and physcion were 119.9-2876 ng(r²=1.0000), 31.48-1007ng(r²=1.0000),17.28-1728ng(r²=1.0000),41.84-1339ng(r²=1.0000) and 5.39-172.5ng(r²=0.9998),respectively.The average recoveries were 100.5%(RSD =2.6%),96.0%(RSD=0.6%),97.8%(RSD=1.5%),97.9%(RSD=1.1%) and 98.1% (RSD=1.6%),respectively.A HPLC fingerprint was developed based on 21 batches of Polygonum cuspidatum. There were 31 common peaks in the fingerprint and 5 were identified.In the meantime, the irradiation of Polygonum cuspidatum sample solution,polydatin solution and resveratrol solution were tested.The contents of polydatin and resveratrol in solutions reduced obviously.Some of them changed into their isomers.RP-HPLC method was developed for the simultaneous determination of polydatin and resveratrol in rat plasma.After being deproteinized by methanol,the plasma samples were analyzed.The assay was shown to be linear over the range of 0.263-33.68μg·mL^-1(r=0.9999) for polydatin and 0.059-37.43μg·mL^-1(r=0.9998) for resveratrol.Mean recoveries were 99.9%(RSD=1.8%) and 93.9%(RSD=4.1%), respectively.The HPLC method developed has been applied to determine the pharmacokinetics of resveratrol in rat plasma after having taken resveratrol orally.The pharmacokinetic parameters were calculated.The time for peak plasma level(T_max) was 27 min and the peak plasma level(C_max) was 1.159μg·mL^-1.The area under concentration-time curve(AUC_0-∞) was 165.2μg·min·mL^-1.RP-HPLC method was developed for the simultaneous determination of polydatin and resveratrol in rat viscus.After being deproteinized by methanol,the viscus samples were analyzed.The assay was shown to be linear over the range of 0.070～35.96μg·mL^-1 (heart r²=0.9996,liver r²=0.9996,spleen r²=0.9984,lung r²=0.9997,kidney r²= 0.9995,brain r²=0.9984) for polydatin and 0.031～31.48μg·mL^-1(heart r²=0.9994, liver r²=0.9995,spleen r²=0.9996,lung r²=1.0000,kidney r²=0.9994,brain r²= 0.9985)) for resveratrol.Mean recoveries of heart were 102.3%(RSD=3.6%) and 102.3%(RSD=5.2%),respectively.Mean recoveries of liver were 101.6%(RSD=3.7%) and 101.7%(RSD=4.8%),respectively.Mean recoveries of spleen were 101.3%(RSD= 4.0%) and 101.0%(RSD=4.4%),respectively.Mean recoveries of lung were 105.8% (RSD=4.6%) and 106.4%(RSD=5.2%),respectively.Mean recoveries of kidney were 100.4%(RSD=3.1%) and 100.0%(RSD=3.4%),respectively.Mean recoveries of brain were 101.6%(RSD=3.2%) and 101.0%(RSD=2.9%),respectively.The HPLC method developed has been applied to determine the distribution of resveratrol in rat viscera after having taken resveratrol orally.The concentrations of viscus except brain were higher than plasma.They reached a high in 20min for heart,spleen,lung,brain,and in 90min for liver and kidney.Then,they went down.The most of resveratrol were collected in lung and liver.LC/MS/MS method and metabolite ID software was applied to study metabolites of rat urine.The experimental result indicates that resveratrol was mainly excreted directly from rat urine as prototype and resveratrol glucuronide and resveratrol sulfate. Polydatin's metabolites were prototype,resveratrol,resveratrol glucuronide and resveratrol sulfate.Their biotransformations in the bodies were bothâ…¡phase metabolism.RP-HPLC method was developed for the simultaneous determination of polydatin and resveratrol in rat excrement.After being treated with C₁₈ Solid Phase Extraction(SPE), the samples were analyzed.The assay was shown to be linear over the range of 0.803-642.6 ng(r=1.0000) for polydatin and 0.814-325.8ng(r=1.0000) for resveratrol.Mean recoveries were 102.2%(RSD=4.3%) and 97.3%(RSD=6.5%),respectively.In the meantime,studied the metabolic transforming of polydatin exerted by rat intestinal bacteria in vitro and in vivo.Incubation experiment in vitro showed that 51%of polydatin was transformed to resveratrol in 4h and 87%in 6h.Through transfer in the stomach and intestine,in vivo experiment displayed that polydatin's relative content was decreased greatly and the resveratrol was relatively increased.It was suggested that polydatin can be metabolized by rat intestinal bacteria and can be transformed into resveratrol through desugarization.The anti-tumour activity in vitro of polydatin,resveratrol and extracter of Polygonum cuspidatum were reported in this dissertation.The studies demonstrated that reaction of reaveratrol was stronger than extracter of Polygonum cuspidatum in inhibiting the proliferation of human A549 cells.Polydatin was almost empty of the reaction.For human HL-60 cells and HO8910 cells,reactions of resveratrol were as strong as extracter of Polygonum cuspidatum,and polydatin was weak.In inhibiting the proliferation of human MCF-7 cells,reaction of resveratrol was better than extracter of Polygonum cuspidatum,and polydatin was weak,too.The studies suggested that polydatin was firstly metabolized into resveratrol in intestines.Then resveratrol was absorbed and partly formed resveratrol glucuronide and resveratrol sulfate.They chiefly distributed in lung and liver.The biotransformations in the bodies wereâ…¡phase metabolism.Partly prototypes and itsâ…¡phase metabolites were excreted from rat urine,and partly prototypes and intestinal metabolites were excreted as stool.