响应面法优化枳椇子多糖提取工艺及其体外生物活性研究
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摘要
目的采用响应面法优化中药枳椇子多糖的提取工艺,并探讨其体外抗氧化和抗黄嘌呤氧化酶(Xanthine oxidase,XO)活性。方法采用中心组合设计(Central composite design,CCD)原理,以料液比、提取时间、提取温度为考察因素,进行3因素5水平的响应面研究。应用DPPH和ABTS法及酶促反应分别测定枳椇子多糖的抗氧化和抗XO活性。结果所得的最佳提取工艺条件为:液料比23 mL·g~(-1),提取时间1.75 h,提取温度86℃。提取的多糖具有明显的清除DPPH和ABTS自由基活性作用,其IC50值分别为143.30μg·mL~(-1)和44.41μg·mL~(-1),体外抑制黄嘌呤氧化酶的IC50为447.33μg·mL~(-1)。结论所优选的工艺条件简便可行,枳椇子多糖具有明显的清除DPPH、ABTS·+自由基活性,以及中等程度的抗XO活性作用,可为中药枳椇子多糖的开发利用和寻找天然抗XO活性分子提供参考。
Objective To optimize the extraction process of polysaccharide from fruits of Hovenia acerba(PHA)by response surface methodology,and to explore its anti-oxidation and anti-xanthine oxidase(XO)activities in vitro.Methods The central composite design(CCD)design was used to study the response surface of 3 factors and 5 levels with liquid-to-solid ratio,extraction time and extraction temperature as factors. The anti-oxidation and antiXO activities of PHA were determined by DPPH and ABTS methods and enzymatic reaction,respectively. Results The optimum extraction conditions were as follows: liquid-to-solid ratio 23 mL·g~(-1), extraction time 1.75 h,extraction temperature 86°C. The resulted polysaccharides had obvious DPPH and ABTS free radical scavenging activities, and their IC50 values were 143.30 μg·mL~(-1) and 44.41 μg·mL~(-1),respectively. The IC50 for inhibiting xanthine oxidase in vitro was 447.33 μg·mL~(-1). Conclusion The optimized extraction conditions were simple and feasible. The PHA had obvious DPPH,ABTS free radical scavenging and moderate anti-XO activities. This study could provide reference for the development and utilization of PHA and the search for natural anti-XO active constituents.
Objective To optimize the extraction process of polysaccharide from fruits of Hovenia acerba(PHA)by response surface methodology,and to explore its anti-oxidation and anti-xanthine oxidase(XO)activities in vitro.Methods The central composite design(CCD)design was used to study the response surface of 3 factors and 5 levels with liquid-to-solid ratio,extraction time and extraction temperature as factors. The anti-oxidation and antiXO activities of PHA were determined by DPPH and ABTS methods and enzymatic reaction,respectively. Results The optimum extraction conditions were as follows: liquid-to-solid ratio 23 mL·g~(-1), extraction time 1.75 h,extraction temperature 86°C. The resulted polysaccharides had obvious DPPH and ABTS free radical scavenging activities, and their IC50 values were 143.30 μg·mL~(-1) and 44.41 μg·mL~(-1),respectively. The IC50 for inhibiting xanthine oxidase in vitro was 447.33 μg·mL~(-1). Conclusion The optimized extraction conditions were simple and feasible. The PHA had obvious DPPH,ABTS free radical scavenging and moderate anti-XO activities. This study could provide reference for the development and utilization of PHA and the search for natural anti-XO active constituents.
引文
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[14]XU H B,YANG T H,XIE P,et al.LC-MS guided isolation of gracilistones A and B,a pair of diastereomeric sesquiterpenoids with an unusual tetrahydrofuran-fused tricyclic skeleton from Acanthopanax gracilistylus and their potential anti-inflammatory activities[J].Fitoterapia,2018,130:265-271.
[15]徐运飞,刘琴,宋珅,等.响应面法优化黄参多糖的提取工艺及其体外抗氧化活性[J].天然产物研究与开发,2015,27(12):2116-2123.
[16]蔡小辉,曾棋平,王艺红,等.星点设计-效应面法优化镰形棘豆多糖的提取工艺[J].药学服务与研究,2017,17(6):427-431.
[17]张海全,钟晓坤,黄勤英,等.响应面法优化苦丁茶熊果酸提取工艺及其抑菌活性研究[J].天然产物研究与开发,2018,30(11):1978-1985.
[18]王会,郭立,谢文磊.抗氧化剂抗氧化活性的测定方法(一)[J].食品与发酵工业,2006,32(3):92-98.
[19]ENROTH C,EGER B T,OKAMOTO K,et al.Crystal structures of bovine milk xanthine dehydrogenase and xanthine oxidase:structure-based mechanism of conversion[J].Proceedings of the National Academy of sciences of the united states of America,2000,97:10723-10728.
[20]许洪波,周瑞,谢培,等.27种中草药的黄嘌呤氧化酶抑制活性筛选[J].时珍国医国药,2017,28(3):547-548.
[21]LIU X,CHEN R,SHANG Y,et al.Superoxide radicals scavenging and xanthine oxidase inhibitory activity of magnesium lithospermate B from Salvia miltiorrhiza[J].Journal of Enzyme Inhibition and Medicinal Chemistry,2009,24:663-668.
[22]ZHU Y,PANDYA B J,CHOI H K.Comorbidities of gout and hyperuricemia in the US general population:NHANES 2007-2008[J].American Journal of Medicine,2012,125(7):679-687.
[2]KAWAI K I,AKIYAMA T,OGIHARA Y,et al.A new sapogenin in the saponins of Zizyphus jujuba,Hovenia dulcis and Bacopa monniera[J].Phytochemistry,1974,13(12):2829-2832.
[3]YOSHIKAWA K,TUMURA S,YAMADA K,et al.Antisweet natural products.vii.hodulosides i,ii,iii,iv,and v from the leaves of Hovenia dulcis Thunb[J].Chemical and Pharmaceutical Bulletin,1992,40(9):2287-2291.
[4]KIMURA Y,KOBAYASHI Y,TAKEDA T,et al.Three new saponins from the leaves of Hovenia dulcis(Rhamnaceae)[J].Journal of the Chemical Society,Perkin Transactions 1,https://doi.org/10.1002/chin.198143305.
[5]PARK J S,KIM I S,REHMAN S U,et al.HPLC determination of bioactive flavonoids in Hovenia dulcis fruit extracts[J].Journal of Chromatographic Science,2015,54(2):130-135.
[6]TAKAI M,OGIHARA Y,SHIBATA S.New peptide alkaloids from Hovenia dulcis and H.tomentella[J].Phytochemistry,1973,12(12):2985-2986.
[7]CHO J Y,MOON J H,EUN J B,et al.Isolation and characterization of 3(Z)-dodecenedioic acid as an antibacterial substance from Hovenia dulcis Thunb[J].Food Science and Biotechnology,2004,13(1):46-50.
[8]贾春晓,熊卫东,毛多斌,等.拐枣果梗中有机酸成分的GC-MS分析[J].中国食品学报,2005,5(1):75-77.
[9]张晶,李慧萍,弓晓杰,等.枳椇属植物化学成分及生物活性研究进展[J].天然产物研究与开发,2006,18(4):674-680.
[10]蔡冰洁,汪苗苗,刘咏.枳椇多糖的分级醇沉及其免疫调节活性的研究[J].合肥工业大学学报(自然科学版),2018,41(6):852-858.
[11]叶文斌,樊亮,王昱,等.拐枣多糖对糖尿病小鼠血糖血脂的影响[J].现代食品科技,2016,32(1):6-12,5.
[12]汪名春.枳椇肉质果梗多糖的分离纯化、结构分析及生物活性研究[D].南京:南京农业大学,2011.
[13]冯沼润,何先元,欧阳凯,等.响应面法优化天葵子总生物碱提取工艺及其体外抗氧化活性研究[J].中药新药与临床药理,2018,29(2):199-204.
[14]XU H B,YANG T H,XIE P,et al.LC-MS guided isolation of gracilistones A and B,a pair of diastereomeric sesquiterpenoids with an unusual tetrahydrofuran-fused tricyclic skeleton from Acanthopanax gracilistylus and their potential anti-inflammatory activities[J].Fitoterapia,2018,130:265-271.
[15]徐运飞,刘琴,宋珅,等.响应面法优化黄参多糖的提取工艺及其体外抗氧化活性[J].天然产物研究与开发,2015,27(12):2116-2123.
[16]蔡小辉,曾棋平,王艺红,等.星点设计-效应面法优化镰形棘豆多糖的提取工艺[J].药学服务与研究,2017,17(6):427-431.
[17]张海全,钟晓坤,黄勤英,等.响应面法优化苦丁茶熊果酸提取工艺及其抑菌活性研究[J].天然产物研究与开发,2018,30(11):1978-1985.
[18]王会,郭立,谢文磊.抗氧化剂抗氧化活性的测定方法(一)[J].食品与发酵工业,2006,32(3):92-98.
[19]ENROTH C,EGER B T,OKAMOTO K,et al.Crystal structures of bovine milk xanthine dehydrogenase and xanthine oxidase:structure-based mechanism of conversion[J].Proceedings of the National Academy of sciences of the united states of America,2000,97:10723-10728.
[20]许洪波,周瑞,谢培,等.27种中草药的黄嘌呤氧化酶抑制活性筛选[J].时珍国医国药,2017,28(3):547-548.
[21]LIU X,CHEN R,SHANG Y,et al.Superoxide radicals scavenging and xanthine oxidase inhibitory activity of magnesium lithospermate B from Salvia miltiorrhiza[J].Journal of Enzyme Inhibition and Medicinal Chemistry,2009,24:663-668.
[22]ZHU Y,PANDYA B J,CHOI H K.Comorbidities of gout and hyperuricemia in the US general population:NHANES 2007-2008[J].American Journal of Medicine,2012,125(7):679-687.