拐枣中总黄酮提取工艺优化及其抗黄嘌呤氧化酶活性研究
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摘要
为了研究拐枣总黄酮(TF)最佳提取工艺及其体外抗黄嘌呤氧化酶(xanthine oxidase,XO)活性。在单因素试验基础上,实验选择料液比、提取时间和乙醇浓度为自变量,应用Box-Benhnken中心组合法进行3因素3水平试验设计,以拐枣总黄酮得率为响应值,进行响应面分析,并采用D101大孔树脂对总黄酮进行进一步富集纯化,在此基础上研究拐枣总黄酮体外抗XO活性。结果表明,拐枣总黄酮的最佳提取条件为料液比15.0 mL/g、提取时间1.85 h、乙醇浓度53.0%,在此条件下,拐枣总黄酮的平均提取率可达到2.56%。体外活性结果表明,我们首次发现拐枣总黄酮具有显著的抗XO活性,其IC_(50 )为18.63±1.67μg/mL。本研究可为从拐枣中寻找和发现天然抗XO活性分子提供参考,同时可为拐枣的开发利用提供思路。
In order to optimize extraction of total flavonoids from Hovenia acerba and investigate xanthine oxidase inhibitory activity.Based on the results of single-factor tests,the ratio of liquid to solid,extraction time and ethanol concentration were optimized by response surface analysis.The results showed that the process of optimum extraction were the ratio of liquid to solid of 15:1(mL:g),ethanol concentration of 53%,extraction time of 1.85 h.Under these conditions,the extraction yield of total flavonoids was 2.56%,which was close to the predicted value.The total flavonoid(TF) was further purified by D101 macroporous resin and evaluated for its xanthine oxidase(XO) inhibitory activities in vitro,and the results showed that TF possessed significant enzyme inhibitory action with IC_(50) value of 18.63 ± 1.67 μg/mL.These results indicated that the optimized extraction technology was reliable and provided a reference for further development and utilization of the Hovenia acerba.
In order to optimize extraction of total flavonoids from Hovenia acerba and investigate xanthine oxidase inhibitory activity.Based on the results of single-factor tests,the ratio of liquid to solid,extraction time and ethanol concentration were optimized by response surface analysis.The results showed that the process of optimum extraction were the ratio of liquid to solid of 15:1(mL:g),ethanol concentration of 53%,extraction time of 1.85 h.Under these conditions,the extraction yield of total flavonoids was 2.56%,which was close to the predicted value.The total flavonoid(TF) was further purified by D101 macroporous resin and evaluated for its xanthine oxidase(XO) inhibitory activities in vitro,and the results showed that TF possessed significant enzyme inhibitory action with IC_(50) value of 18.63 ± 1.67 μg/mL.These results indicated that the optimized extraction technology was reliable and provided a reference for further development and utilization of the Hovenia acerba.
引文
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2 Zhang B,Li LT,Zhang M,et al.Optimization of extraction technology of Hovenia acerba sticks polysaccharides and its antioxidant activity[J].China Brew(中国酿造),2016,35:155-160.
3 Ren QL,Lin B,Zhou Y,et al.Therapeutic effect of Hovenia acerba extract on renal calculi in rats[J].China J Mod Med(中国现代医学杂志),2017,27:24-27.
4 Zhang XQ,Xu FF,Wang L,et al.Two pairs of new diastereoisomeric flavonolignans from the seeds of Hovenia acerba[J].Phytochem Lett,2012,5:292-296.
5 Xu FF,Zhang XQ,Zhang J,et al.Two methyl-migrated 16,17-seco-dammarane triterpenoid saponins from the seeds of Hovenia acerba[J].J Asian Nat Prod Res,2012,14:135-140.
6 Zhang J,Li HP,Gong XJ,et al.Research advances in chemical constituents of the genus Hovenia and their pharmacological activities[J].Nat Prod Res Dev(天然产物研究与开发),2006,18:674-680,666.
7 Wen W,Zhang H,Zeng R.Optimization of extracting conditions of Zhijuzi by alcohol[J].Herald Med(医药导报),2008,27:77-79.
8 Tian ZY,Chai Z,Xu YF,et al.Optimizing the ultrasonic extracting method of total flavonoids from Hovenia dulcis Thunb by orthogonal design[J].J Henan Univ:Med Ed(河南大学学报:医学版),2015,34:247-251.
9 Xu HB,Zhou R,Xie P,et al.Xanthine oxidase inhibitory activity of some Chinese medicinal plants[J].Lishizhen Med Mater Med Res(时珍国医国药),2017,28:547-548.
10 Zhu XP,Yu MM,Feng J,et al.Study on stability of total flavonoids from Hovenia dulcis Thumb[J].Food Fer Sci&Tech(食品与发酵科技),2017,53:68-71.
11 Wang ZC,Jiang YR,Liu XW,et al.Optimization of extraction of total flavonoids from zingiber officinale Roscoe stem and leaves using response surface methodology and evaluation of its antioxidant activity[J].Nat Prod Res Dev(天然产物研究与开发),2015,27:1582-1588.
12 Enroth C,Eger BT,Okamoto K,et al.Crystal structures of bovine milk xanthine dehydrogenase and xanthine oxidase:structure-based mechanism of conversion[J].Proc Natl Acad Sci,2000,97:10723-10728.
13 Kong LD,Cai Y,Huang WW,et al.Inhibition of xanthine oxidase by some Chinese medicinal plants used to treat gout[J].J Ethnopharmacol,2000,73:199-207.
14 Liu X,Chen R,Shang Y,et al.Superoxide radicals scavenging and xanthine oxidase inhibitory activity of magnesium lithospermate B from Salvia miltiorrhiza[J].J Enzyme Inhib Med Chem,2009,24:663-668.
15 Zhu Y,Pandya BJ,Choi HK.Comorbidities of gout and hyperuricemia in the US general population:NHANES 2007-2008[J].Am J Med,2012,125:679-687.