UPLC同时测定野菊花药材中10个黄酮和有机酸的含量
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摘要
目的:建立UPLC同时测定野菊花药材中绿原酸、咖啡酸、1,3-二咖啡酰奎宁酸、木犀草素-7-O-β-D-葡萄糖苷、3,4-二咖啡酰奎宁酸、3,5-二咖啡酰奎宁酸、4,5-二咖啡酰奎宁酸、木犀草素、蒙花苷及芹菜素10个成分含量的方法。方法:采用Agilent Eclipse Plus C_(18)色谱柱(2.1 mm×100 mm,1.8μm);以甲醇(A)-0.3%磷酸水溶液(B)为流动相,梯度洗脱,流速0.4 mL·min~(-1);检测波长326 nm;柱温25℃。结果:10个成分在38 min内均达到基线分离,线性关系良好(r>0.999 2,n=6);平均回收率均在96.4%～103.2%(RSD<2.0%,n=6);测定的6批样品中上述10个成分含量范围分别为1.018～3.430、0.079～0.365、0～0.269、1.198～2.754、0.185～0.765、1.135～6.659、0.587～2.808、0.103～1.663、0.495～13.050、0.214～0.933 mg·g~(-1)。结论:本方法操作简单,准确性和重复性良好。可用于野菊花药材的多成分同步测定,为野菊花的全面质量评价和质量控制提供科学依据。
Objective:To establish a ultra-performance liquid chromatography(UPLC)method for the simultaneous determination of chlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,luteolin-7-O-β-D-glucoside,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,luteolin,linarin and apigenin in Chrysanthemum indicum. Methods:The separation was carried out on an Agilent Eclipse Plus C_(18)(2.1 mm×100 mm,1.8 μm)column eluted with methanol(A)and 0.3%phosphoric acid(B)with gradient mode at a flow rate of 0.4 mL·min~(-1). The column temperature was 25 ℃and the detection wavelength was set at 326 nm. Results:Good separation of chlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,luteolin-7-O-β-D-glucoside,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,luteolin,linarin and apigenin was achieved within 38 min. Calibration curves of the ten components showed good linear relationship(r>0. 999 2,n=6). The average recoveries were within 96.4%-103.2%(RSD<2.0%,n=6). The content ranges of the above mentioned 10 components in 6 batches of samples were 1.018-3.430,0.079-0.365,0-0.269,1.198-2.754,0.185-0.765,1.135-6.659,0.587-2.808,0.103-1.663,0.495-13.050 and 0.214-0.933 mg·g~(-1),respectively. Conclusion:The method is simple,accurate and repeatable and can be used for simultaneous determination of multi-components in Chrysanthemum indicum,which provides scientific base for comprehensive evaluation and quality control of the herb.
Objective:To establish a ultra-performance liquid chromatography(UPLC)method for the simultaneous determination of chlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,luteolin-7-O-β-D-glucoside,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,luteolin,linarin and apigenin in Chrysanthemum indicum. Methods:The separation was carried out on an Agilent Eclipse Plus C_(18)(2.1 mm×100 mm,1.8 μm)column eluted with methanol(A)and 0.3%phosphoric acid(B)with gradient mode at a flow rate of 0.4 mL·min~(-1). The column temperature was 25 ℃and the detection wavelength was set at 326 nm. Results:Good separation of chlorogenic acid,caffeic acid,1,3-dicaffeoylquinic acid,luteolin-7-O-β-D-glucoside,3,4-dicaffeoylquinic acid,3,5-dicaffeoylquinic acid,4,5-dicaffeoylquinic acid,luteolin,linarin and apigenin was achieved within 38 min. Calibration curves of the ten components showed good linear relationship(r>0. 999 2,n=6). The average recoveries were within 96.4%-103.2%(RSD<2.0%,n=6). The content ranges of the above mentioned 10 components in 6 batches of samples were 1.018-3.430,0.079-0.365,0-0.269,1.198-2.754,0.185-0.765,1.135-6.659,0.587-2.808,0.103-1.663,0.495-13.050 and 0.214-0.933 mg·g~(-1),respectively. Conclusion:The method is simple,accurate and repeatable and can be used for simultaneous determination of multi-components in Chrysanthemum indicum,which provides scientific base for comprehensive evaluation and quality control of the herb.
引文
[1]王志东,梁容瑞,李宗芳.中药野菊花的药理作用研究进展[J].医学综述,2009,15(6):906WANG ZD,LIANG RR,LI ZF.Development in pharmacological research of Chrysanthemum indicum[J].Med Recapit,2009,15(6):906
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[11]夏伯候,周亚敏,谢家驰,等.UPLC法测定野菊花中5种抗氧化活性成分的含量[J].中药材,2016,39(2):348XIA BH,ZHOU YM,XIE JC,et al.Determination dients contents of Flos Chrysanthemi Indici by UPLC[J].J Chin Med Mater,2016,39(2):348
[12]吴明侠,王晶娟,张贵君.野菊花水煎剂中7种药效组分的含量测定[J].中成药,2011,33(2):300WU MX,WANG JJ,ZHANG GJ.Determination of the seven active components in water extraction of Chrysanthemi Indici Flos[J].Chin Tradit Pat Med,2011,33(2):300
[13]沈亚芬,杜伟锋,郑敏霞,等.HPLC同时测定野菊花中绿原酸、咖啡酸和蒙花苷的含量[J].中国现代应用药学,2015,32(9):1117SHEN YF,DU WF,ZHENG MX,et al.Simultaneous determination of chlorogenic acid,caffeic acid and linarin in Chrysanthemi Indici Flos by HPLC[J].Chin J Mod Appl Pharm,2015,32(9):1117
[14]申海进,郭巧生,房海灵,等.RP-HPLC测定野菊花中槲皮素、木犀草素、芹菜素和刺槐素[J].中国中药杂志,2010,35(2):191SHEN HJ,GUO QS,FANG HL,et al.Determination of quercetin,luteolin,apigenin and acacetin in Flos Chrysanthemi Indici by RP-HPLC[J].China J Chin Mater Med,2010,35(2):191
[15]张明.野菊花抗氧化活性成分分离、鉴定及初步分析的研究[D].合肥:安徽医科大学,2011ZHANG M.Isolation,Identification and Analysis of Antioxidant Constituents in Chrysanthemum indicum L.[D].Hefei:Anhui Medical University,2011
[16]符玲,潘成学,蒋莹,等.不同产地的市售野菊花中绿原酸的含量测定[J].中国实验方剂学杂志,2010,16(5):68FU L,PAN CX,JIANG Y,et al.Determination of chlorogenic acid in Chrysanthemum indicum in different habitats[J].Chin J Exp Tradit Med Form,2010,16(5):68
[17]刘菲,杨洋,谭晓杰,等.RP-HPLC同时测定野菊花中6种黄酮的含量[J].中国中药杂志,2009,34(16):2067LIU F,YANG Y,TANG XJ,et al.Simultaneous determination of six flavonoids in Flos Chrysanthemi Indici by RP-HPLC[J].China J Chin Mater Med,2009,34(16):2067
[2]张骏艳,张磊,李俊,等.野菊花总黄酮对佐剂性关节炎大鼠氧自由基代谢的影响[J].中国中药杂志,2010,35(3):344ZHANG JY,ZHANG L,LI J,et al.Effects of total flavonoids of Chrysanthemum indicum on free radical in adjuvant arthritic rats[J].China J Chin Mater Med,2010,35(3):344
[3]吴钉红,杨立伟,苏薇薇,等.野菊花化学成分及药理研究进展[J].中药材,2004,27(2):143WU DH,YANG LW,SU WW,et al.Advances in study on chemical contituents and pharmacological effects of Chrysanthemum indicum[J].J Chin Med Mater,2004,27(2):143
[4]陈传千,沈艳平,屈跃丹,等.野菊花提取物药理作用的研究进展[J].吉林医药学院学报,2010,31(3):175CHEN CQ,SHEN YP,QU YD,et al.Study progress of pharmacological actions of Chrysanthemum indicum extract[J].JJilin Med Coll,2010,31(3):175
[5]宋立人,洪恂,丁绪亮,等.现代中药学大辞典[M].北京:人民卫生出版社,2001:1968SONG LR,HONG X,DING XL,et al.Comprehensive Dictionary of Modern Traditional Chinese Medicine[M].Beijing:People’s Medical Publishing House,2001:1968
[6]陈耕夫,郭晓玲,孟青.野菊花化学成分分析[J].中药材,2002,25(2):103CHEN GF,GUO XL,MENG Q.Studie on chemical contituents of Chrysanthemum indicum[J].J Chin Med Mater,2002,25(2):103
[7]吴雪松,许浚,张铁军,等.野菊的化学成分及质量评价研究进展[J].中草药,2015,6(3):43WU XS,XU J,ZHANG TJ,et al.Research progress in chemical constituents of Chrysanthemum indicum and their quality assessment[J].Chin Tradit Herb Drugs,2015,46(3):443
[8]裴姗姗,毕跃峰,田野,等.野菊花的研究进展[J].河南中医学院学报,2007,22(6):83PEI SS,BI YF,TIAN Y,et al.Investigation advancement of the wild Chrysanthemum[J].J Henan Univ Chin Med,2007,22(6):83
[9]赵昱,赵军,李湘萍,等.咖啡酰奎尼酸类化合物研究进展[J].中国中药杂志,2006,31(11):869ZHAO Y,ZHAO J,LI XP,et al.Advances in caffeoylquinic acid research[J].China J Chin Mater Med,2006,31(11):869
[10]万玉珊,高雅婷,张莹,等.D-奎尼酸研究进展[J].黑龙江医药,2013,26(5):819WANG YS,GAO YT,ZHANG Y,et al.Progress in researches for D-quinic acid[J].Heilongjiang Med J,2013,26(5):819
[11]夏伯候,周亚敏,谢家驰,等.UPLC法测定野菊花中5种抗氧化活性成分的含量[J].中药材,2016,39(2):348XIA BH,ZHOU YM,XIE JC,et al.Determination dients contents of Flos Chrysanthemi Indici by UPLC[J].J Chin Med Mater,2016,39(2):348
[12]吴明侠,王晶娟,张贵君.野菊花水煎剂中7种药效组分的含量测定[J].中成药,2011,33(2):300WU MX,WANG JJ,ZHANG GJ.Determination of the seven active components in water extraction of Chrysanthemi Indici Flos[J].Chin Tradit Pat Med,2011,33(2):300
[13]沈亚芬,杜伟锋,郑敏霞,等.HPLC同时测定野菊花中绿原酸、咖啡酸和蒙花苷的含量[J].中国现代应用药学,2015,32(9):1117SHEN YF,DU WF,ZHENG MX,et al.Simultaneous determination of chlorogenic acid,caffeic acid and linarin in Chrysanthemi Indici Flos by HPLC[J].Chin J Mod Appl Pharm,2015,32(9):1117
[14]申海进,郭巧生,房海灵,等.RP-HPLC测定野菊花中槲皮素、木犀草素、芹菜素和刺槐素[J].中国中药杂志,2010,35(2):191SHEN HJ,GUO QS,FANG HL,et al.Determination of quercetin,luteolin,apigenin and acacetin in Flos Chrysanthemi Indici by RP-HPLC[J].China J Chin Mater Med,2010,35(2):191
[15]张明.野菊花抗氧化活性成分分离、鉴定及初步分析的研究[D].合肥:安徽医科大学,2011ZHANG M.Isolation,Identification and Analysis of Antioxidant Constituents in Chrysanthemum indicum L.[D].Hefei:Anhui Medical University,2011
[16]符玲,潘成学,蒋莹,等.不同产地的市售野菊花中绿原酸的含量测定[J].中国实验方剂学杂志,2010,16(5):68FU L,PAN CX,JIANG Y,et al.Determination of chlorogenic acid in Chrysanthemum indicum in different habitats[J].Chin J Exp Tradit Med Form,2010,16(5):68
[17]刘菲,杨洋,谭晓杰,等.RP-HPLC同时测定野菊花中6种黄酮的含量[J].中国中药杂志,2009,34(16):2067LIU F,YANG Y,TANG XJ,et al.Simultaneous determination of six flavonoids in Flos Chrysanthemi Indici by RP-HPLC[J].China J Chin Mater Med,2009,34(16):2067