HPLC同时测定大叶冬青叶中10种多酚成分的含量及抗氧化活性研究
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摘要
建立HPLC同时测定大叶冬青叶中新绿原酸、绿原酸、隐绿原酸、咖啡酸、对羟基肉桂酸、芦丁、异槲皮苷、异绿原酸B、异绿原酸A和异绿原酸C 10种多酚类化合物的定量分析方法,研究不同产地及不同采收时期大叶冬青叶中10种多酚类化合物的含量差异及其抗氧化活性。采用HPLC测定大叶冬青叶50%甲醇提取液中10种多酚类成分的含量,并选用DPPH法对其抗氧化活性进行初步探索。结果表明,10种多酚类化合物分离效果较好,标准曲线在检测范围内具有良好的线性(r>0.999 5),平均加样回收率在96.58%～112.03%,RSD<4%(n=6)。大叶冬青叶50%甲醇提取液抗氧活性良好。本实验建立的方法快速、准确、重复性好,可用于同时测定大叶冬青叶中新绿原酸、绿原酸、隐绿原酸、咖啡酸、对羟基肉桂酸、芦丁、异槲皮苷、异绿原酸B、异绿原酸A、异绿原酸C 10种成分的含量;不同产地、不同采收时期大叶冬青叶中10种多酚类化合物含量存在一定差异;不同来源大叶冬青叶50%甲醇提取液抗氧化能力具有明显差异。
This research established an HPLC method for determination of ten kinds of polyphenol including the neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,p-hydroxycinnamic acid,rutin,isoquercitrin,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the leaves of Ilex latifolia,and investigated the content difference and the antioxidant activity.Determination of 10 polyphenols in 50% methanol extract from the leaves of I.latifolia by HPLC and preliminary study on its antioxidant activity by DPPH.The results showed that Ten compounds were well separated with good linear correlations(r>0.999 5).The average recoveries of ten compounds were 96.58%-112.03% and the RSD were less than 4%(n=6).The 50% methanol extract from the leaves of I.latifolia has good antioxidant activity.The HPLC method was rapid,accurate and stable,which could be used to detect simultaneously neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,p-hydroxycinnamic acid,rutin,isoquercitrin,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the leaves of I.latifolia.There are certain differences in the contents of ten compounds between the samples from different producing areas and harvest times.The antioxidant capacity of 50% methanol extracts from the leaves of I.latifolia from different sources is significantly different.
This research established an HPLC method for determination of ten kinds of polyphenol including the neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,p-hydroxycinnamic acid,rutin,isoquercitrin,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the leaves of Ilex latifolia,and investigated the content difference and the antioxidant activity.Determination of 10 polyphenols in 50% methanol extract from the leaves of I.latifolia by HPLC and preliminary study on its antioxidant activity by DPPH.The results showed that Ten compounds were well separated with good linear correlations(r>0.999 5).The average recoveries of ten compounds were 96.58%-112.03% and the RSD were less than 4%(n=6).The 50% methanol extract from the leaves of I.latifolia has good antioxidant activity.The HPLC method was rapid,accurate and stable,which could be used to detect simultaneously neochlorogenic acid,chlorogenic acid,cryptochlorogenic acid,caffeic acid,p-hydroxycinnamic acid,rutin,isoquercitrin,isochlorogenic acid B,isochlorogenic acid A and isochlorogenic acid C in the leaves of I.latifolia.There are certain differences in the contents of ten compounds between the samples from different producing areas and harvest times.The antioxidant capacity of 50% methanol extracts from the leaves of I.latifolia from different sources is significantly different.
引文
1 Chen JX.Characteristics,uses and main propagation and cultivation techniques of Ilex latifolia[J].Shanghai Agric Sci Tech(上海农业科技),2014,3:99.
2 Wang CQ,Wang L,Fan CL,et al.Ilelic acids A and B,two unusual triterpenes with a seven-membered ring from Ilex latifolia[J].Organic Letters,2012,14:4102-4105.
3 Wang CQ,Li MM,Zhang W,et al.Four new triterpenes and triterpene glycosides from the leaves of Ilex latifolia,and their inhibitory activity on triglyceride accumulation[J].Fitoterapia,2015,106:141-146.
4 Wang CQ,Wang HT,Xu H,et al.Triterpenoids from leaves of Ilex latifolia[J].Chin Med Mat(中药材),2015,38:1654-1656.
5 Wang CQ,Wang L,Li BJ,et al.Study on flavonoids from Ilex latifolia[J].China J Chin Mater Med(中国中药杂志),2014,39:258-261.
6 Wang CQ,Wang HT,Chen J,et al.Research progress on three terpenoids and pharmacological effects of Ilex Kudingcha[J].Chin J Clin Pharmacol Ther(中国临床药理学与治疗学),2016,21:703-709.
7 Wang CQ,Zhang YH.Advances in research on chemical components and pharmacological activities of Ilex latifolia[J].China Pharm(中国药房),2014,25:1052-1054.
8 Fan CL,Fan L,Zhang XQ,et al.Triterpenoid saponins from the leaves of Ilex latifolia[J].Chin Pharm J(中国药学杂志),2010,45:1228-1232.
9 State Administration of Traditional Chinese Medicine Chinese Materia Medica Editorial Board.Chinese herbal medicine:Vol 13(中华本草:第13卷)[M].Shanghai:Shanghai Scientific & Technical Publishers,1999:4032.
10 Jiangsu New Medical College.Dictionary of Chinese medicine:Vol 1(中药大辞典:上册)[M].Shanghai:Shanghai Scientific & Technical Publishers,1977,1288.
11 Wang HT,Wang CQ.27-O-(E)-p-coumaric acyl ursolic acid via JNK /SAPK signal pathway regulates apoptosis of human breast cancer MDA-MB-231 cell line[J].China J Chin Mater Med(中国中药杂志),2015,40:722-726.
12 Fan J,Wu Z,Zhao T,et al.Characterization,antioxidant and hepatoprotective activities of polysaccharides from Ilex latifolia Thunb[J].Carbohydrate Polymers,2014,101:990-997.
13 Zheng W,Michaeln C,Paul S.Analysis of chlorogenic acids in beverages prepared from chinese health foods and investigation,in vitro,of effects on glucose absorption in cultured Caco-2 cells[J].Food Chem,2008,108:369-373.
14 Chen JL.Studies on the extraction and bioactivity of kuding tea polyphenols[D].Guangzhou:South China Agricultural University,2016.
15 Hu T.Study on isolation and identification of Ilex latifolia Thunb and its bioactivities[D].Guangzhou:South China University of Technology(华南理工大学),2013.
16 Sun Y.Extraction,purification,structure and antionxidant activities of polyphenols and polysaccharides in Ilex kudingcha[D].Nanjing:Nanjing Agricultural University(南京农业大学),2010.
17 Guo Y.Study on the antioxidant activity of six Kudingcha species in aquifoliaceae[D].Haikou:Hainan University(海南大学),2013.
18 Chen W,Wang HS,Deng YX,et al.Study on the extraction and antioxidant activity of the components of Ilex kudingcha[J].Nat Prod Res Dev(天然产物研究与开发),2002,14:24-28.
19 Sharma OP,Bhat TK.DPPH antioxidant assay revisited[J].Food Chem,2009,113:1202-1205.
20 Feng YH,Li C,Xin WM,et al.Exploration on feasibility of introducing bioassay method into quality evaluation of chinese herbal medicines by studying on the correlation between antioxidant activity of prunella vulgaris and its total phenolic acids content for example[J].China J Chin Mater Med(中国中药杂志),2016,41:2660-2668.
2 Wang CQ,Wang L,Fan CL,et al.Ilelic acids A and B,two unusual triterpenes with a seven-membered ring from Ilex latifolia[J].Organic Letters,2012,14:4102-4105.
3 Wang CQ,Li MM,Zhang W,et al.Four new triterpenes and triterpene glycosides from the leaves of Ilex latifolia,and their inhibitory activity on triglyceride accumulation[J].Fitoterapia,2015,106:141-146.
4 Wang CQ,Wang HT,Xu H,et al.Triterpenoids from leaves of Ilex latifolia[J].Chin Med Mat(中药材),2015,38:1654-1656.
5 Wang CQ,Wang L,Li BJ,et al.Study on flavonoids from Ilex latifolia[J].China J Chin Mater Med(中国中药杂志),2014,39:258-261.
6 Wang CQ,Wang HT,Chen J,et al.Research progress on three terpenoids and pharmacological effects of Ilex Kudingcha[J].Chin J Clin Pharmacol Ther(中国临床药理学与治疗学),2016,21:703-709.
7 Wang CQ,Zhang YH.Advances in research on chemical components and pharmacological activities of Ilex latifolia[J].China Pharm(中国药房),2014,25:1052-1054.
8 Fan CL,Fan L,Zhang XQ,et al.Triterpenoid saponins from the leaves of Ilex latifolia[J].Chin Pharm J(中国药学杂志),2010,45:1228-1232.
9 State Administration of Traditional Chinese Medicine Chinese Materia Medica Editorial Board.Chinese herbal medicine:Vol 13(中华本草:第13卷)[M].Shanghai:Shanghai Scientific & Technical Publishers,1999:4032.
10 Jiangsu New Medical College.Dictionary of Chinese medicine:Vol 1(中药大辞典:上册)[M].Shanghai:Shanghai Scientific & Technical Publishers,1977,1288.
11 Wang HT,Wang CQ.27-O-(E)-p-coumaric acyl ursolic acid via JNK /SAPK signal pathway regulates apoptosis of human breast cancer MDA-MB-231 cell line[J].China J Chin Mater Med(中国中药杂志),2015,40:722-726.
12 Fan J,Wu Z,Zhao T,et al.Characterization,antioxidant and hepatoprotective activities of polysaccharides from Ilex latifolia Thunb[J].Carbohydrate Polymers,2014,101:990-997.
13 Zheng W,Michaeln C,Paul S.Analysis of chlorogenic acids in beverages prepared from chinese health foods and investigation,in vitro,of effects on glucose absorption in cultured Caco-2 cells[J].Food Chem,2008,108:369-373.
14 Chen JL.Studies on the extraction and bioactivity of kuding tea polyphenols[D].Guangzhou:South China Agricultural University,2016.
15 Hu T.Study on isolation and identification of Ilex latifolia Thunb and its bioactivities[D].Guangzhou:South China University of Technology(华南理工大学),2013.
16 Sun Y.Extraction,purification,structure and antionxidant activities of polyphenols and polysaccharides in Ilex kudingcha[D].Nanjing:Nanjing Agricultural University(南京农业大学),2010.
17 Guo Y.Study on the antioxidant activity of six Kudingcha species in aquifoliaceae[D].Haikou:Hainan University(海南大学),2013.
18 Chen W,Wang HS,Deng YX,et al.Study on the extraction and antioxidant activity of the components of Ilex kudingcha[J].Nat Prod Res Dev(天然产物研究与开发),2002,14:24-28.
19 Sharma OP,Bhat TK.DPPH antioxidant assay revisited[J].Food Chem,2009,113:1202-1205.
20 Feng YH,Li C,Xin WM,et al.Exploration on feasibility of introducing bioassay method into quality evaluation of chinese herbal medicines by studying on the correlation between antioxidant activity of prunella vulgaris and its total phenolic acids content for example[J].China J Chin Mater Med(中国中药杂志),2016,41:2660-2668.