| Hyperuricemia is characterized by an abnormally high level of uric acid in the blood mainly associated with overproduction and/or underexcretion,an important biochemical basis leading to gout,which is closely related to cardiovascular disease,chronic kidney disease and metabolic syndrome.It's controled by decreasing the formation of uric acid and promoting uric acid excretion.Xanthine oxidase,a key enzyme closely related to the biosynthesis of uric acid,is an important target of drugs used as decreasing the level of uric acid.Research on exploring effective,nontoxic and clear mechanism drugs for the treatment of hyperuricemia has become the popular and difficult point researched by domestic and foreign scholars.In several studies,the researchers evaluated that the rhizome of Polygonum cuspidatum extraction can treat the gout by its strong inhibition of the enzyme xanthine oxidase and significantly reduce serum uric acid level of hyperuricemia rats.Through former researches of our team,the ethanol extraction of Polygonum cuspidatum possesses inhibitory activities.The material foundation and action mechanism has been studyed.We found that torachrysone-8-O-?-D-glucopyranoside was able to induce an immediate decrease in uric acid level.To our best knowledge,there are no reports in the literature showing xanthine oxidase inhibitory activity of this compound.At the same time,emodin-1-O-?-D-glucopyranoside,emodin-8-O-?-D-glucopyranoside and physcion-8-O-?-D-glucopyranoside could be used as potential inhibitories for the inhibition of xanthine oxidase.Piecid and resveratrol expressed decrease in uric acid level based on promoting uric acid excretion.Based on the previous research,we expect to prepare effective ingredients from Polygonum cuspidatum and provide new ideas for the treatment of hyperuricemia under the circumstance of clarity of material basis and mechanism of action.In this research,two reference substance standards were prepared by macroporous resin column chromatography,RP-18 reversed-phase silica gel column,semi-preparative HPLC and high-speed counter-current chromatography.The purity was determined by HPLC analysis using the main component self-compare with no corretion factor and photodiode array detector purity test methods.The study showed that two copounds represented only one main peak by using different chromatographic columns and gradient elutions.The purity of them was determined by HPLC analysis using the main component self-comparison with no correction factor method.The results showed that the purity of these two conpounds was all over 98% which were in accordance with the determination requirement.The extration efficiency was evaluated by the high performance liquid chromatographic quantification of piecid,torachrysone-8-O-?-D-glucopyranoside,emodin-8-O-?-D-glucopyranoside,resveratrol and emodin,which are five main active ingredients in Polygonum cuspidatum.Microwave assisted extraction(MAE)was used to extract these five ingredients.The extraction factors including extration solvents,extration temperature,microwave power and extration time were investigated.The best combination of extration parameters were obtained as followed: PEG 200 solution concentration of 60%,microwave power of 700 W,extration temperation of 80 ?,and the extraction time of 10 min.The correlation coefficients,precisions,stabilities and reproducibilities confirmed the validation of the proposed extracion method.During the separation and purification research,macroporous adsorption resin technique for separating effective ingredients from Polygonum cuspidatum was studied.Six macroporous resins were tested for their adsorption and desorption performance to five ingredients.Static,dynamic adsorption and desorption experiments were conducted.Meanwhile,the adsorption and desorption rates were concerned to test out the prominent macroporous resin and optimal processing parameters.HPD-600 was subsequently selected as the best macroporous resin.The optimal processing parameters were obtained as follows: the loading volume was 60 m L(1 BV),sample solutions(which contained piecid,resveratrol,torachrysone-8-O-?-D-glucopyranoside,emodin-8-O-?-D-glucopyranoside and emodin were 0.7477,0.0688,0.0437,0.2763 and 0.1343 mg·m L-1,respectively)was 2 BV and the solutions were loaded into a glass columnat a flow rate of 1 BV·h-1.The HPD-600 column was washed with distilled water(4 BV),10% ethanol(4 BV)and 90% ethanol(10 BV)at a flow rate of 1 BV·h-1.As a result,the contents of effective ingredients were increased from 21.46% to 31.34% with 86.59% revecory.The method with HPD-600 as the optimal macroporous resin undergoing with the modified purification process is a simple,safe and inexpensive technology,which could be a promising potential of purification from Polygonum cuspidatum.The results of the research can provide scientific evidience for the development of anti-hyperuricemia drugs,as well as lay the foundation for the development of innovative drugs with independent intellectual property rights,which has important academic and applied prospects. |
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