平卧菊三七提取物抗氧化活性研究与抗氧化特征成分的HPLC-MS/MS分析
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摘要
目的:采用DPPH体系测定平卧菊三七地上部分不同极性部位提取物的抗氧化活性,并利用液质联用(HPLC-MS/MS)技术分析了活性较高的乙酸乙酯部位的抗氧化特征性成分。方法:对平卧菊三七地上部分提取物用石油醚、乙酸乙酯、正丁醇进行萃取得到石油醚部位、乙酸乙酯部位、正丁醇部位及水部位。考察平卧菊三七地上部分不同极性部位提取物对清除1,1-二苯基-2-三硝基苯肼(DPPH)自由基的能力以及通过液质联用电喷雾四级杆飞行时间质谱仪(HPLC-ESI-Q-TOF-MS)技术对其活性部位中的化学成分进行定性分析。采用液质联用(LC-MS/MS),C_(18)色谱柱(150 mm×4.6 mm,5μm)以甲醇-0.1%甲酸溶液为流动相梯度洗脱,使用ESI负离子模式下采集数据。通过质谱数据分析,并与对照品及文献对照,对其进行成分鉴定。结果:平卧菊三七地上部分不同极性提取物对DPPH自由基都具有清除力,其中乙酸乙酯部位的清除能力最强。通过其质谱数据分析,并与对照品及相关文献对照,共鉴定出11个化合物。结论:应用DPPH抗氧化活性筛选及HPLC-ESI-Q-TOF-MS技术能够快速准确地发现平卧菊三七地上部分抗氧化活性成分,为进一步研究平卧菊三七的药效物质基础提供参考。
Objective:To identify the antioxidant constituents of Gynura procumbens extract with DPPH radical-scavenging activity and HPLC-MS/MS technology.Methods:The antioxidant activities of different polar parts of Gynura procumbens extract were determined by using the DPPH radical-scavenging activity.And the antioxidant characteristics of EtOAc extract were analyzed by comparing the data with reference substances and the published papers.The separation and identification were carried out by HPLC-MS/MS analysis with an Agilent Hanbang C_(18) column(150 mm×4.6 mm,5 μm) using 0.1% formic acid and methanol as mobile phase.Results:In vitro antioxidant assays showed that the different polar parts of G.procumbens extract displayed varying degrees of efficacy in a dose-dependent manner.EtOAc extract exhibited the strongest DPPH radical-scavenging ability with IC_(50) value of 28.8 μg·mL~(-1).Eleven main constituents in the EtOAc extract were identified by HPLC-MS/MS.Conclusion:DPPH radical-scavenging ability method and HPLC-MS/MS play a huge important role in exploring the underlying antioxidant activity and related substances.
Objective:To identify the antioxidant constituents of Gynura procumbens extract with DPPH radical-scavenging activity and HPLC-MS/MS technology.Methods:The antioxidant activities of different polar parts of Gynura procumbens extract were determined by using the DPPH radical-scavenging activity.And the antioxidant characteristics of EtOAc extract were analyzed by comparing the data with reference substances and the published papers.The separation and identification were carried out by HPLC-MS/MS analysis with an Agilent Hanbang C_(18) column(150 mm×4.6 mm,5 μm) using 0.1% formic acid and methanol as mobile phase.Results:In vitro antioxidant assays showed that the different polar parts of G.procumbens extract displayed varying degrees of efficacy in a dose-dependent manner.EtOAc extract exhibited the strongest DPPH radical-scavenging ability with IC_(50) value of 28.8 μg·mL~(-1).Eleven main constituents in the EtOAc extract were identified by HPLC-MS/MS.Conclusion:DPPH radical-scavenging ability method and HPLC-MS/MS play a huge important role in exploring the underlying antioxidant activity and related substances.
引文
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[2] HEW C S,KHOO B Y,GAM L H.The Anti-Cancer Property of Proteins Extracted from Gynura procumbens (Lour.) Merr[J].Plos One,2013,8(7):1-10.
[3] 门凤麟,王英锋,祝翔.平卧菊三七提取物降血糖实验研究[J].首都师范大学学报(自然科版),2015,36(2):39-42.
[4] LI X J,MU Y M,LI T T,etal.Gynura procumbens Reverses Acute and Chronic Ethanol-Induced Liver Steatosis through MAPK/SREBP-1c-Dependent and-Independent Pathways[J].J Agr Food Chem,2015,63(38):8460-8471.
[5] KIM J,LEE C W,KIM E K,etal.Inhibition effect of Gynura procumbens extract on UV-B-induced matrix-metalloproteinase expression in human dermal fibroblasts[J].J Ethnopharmacol,2011,137(1):427-433.
[6] 何英杰,谢红旗,伍睿宇,等.HPLC-DPPH快速测定枳壳抗氧化活性成分[J].中国现代中药,2017,19(8):1131-1135.
[7] 王增绘,王冬梅,刘艾琳,等.山豆根抑制丁酰胆碱酯酶活性及活性部位UPLC-Q-TOF-MS分析[J].中国现代中药,2015,17(9):912-916.
[8] CHEN J,MANGELINCKX S,LU H,etal.Profiling and Elucidation of the Phenolic Compounds in the Aerial Parts of Gynura bicolor and G.divaricata Collected from Different Chinese Origins[J].Chem Biodivers,2015,12(1):96-115.
[9] 王亚丹,何轶,戴忠,等.HPLC-MS/MS法同时测定山银花中7个有机酸的含量[J].药物分析杂志,2016,36(6):998-1005.
[10] 张阿琴,李迩娜,张仓,等.LC-MS分析消癌平注射液中有机酸类成分[J].中国实验方剂学杂志,2012,18(4):88-90.