建立UPLC-MS联合HPLC方法探究外源物质对三七叶皂苷含量和生长的影响  

Development of UPLC-MS Combined with HPLC Technique to Explore the Effects of Exogenous Substances on the Saponin Contents and Growth of Panax notoginseng Leaves

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作  者:贾兵 范伟 陈庚 李莹 张广辉 杨生超 陈军文 JIA Bing;FAN Wei;CHEN Geng;LI Ying;ZHANG Guanghui;YANG Shengchao;CHEN Junwen(National&Local Joint Engineering Research Center on Germplasms Utilization&Innovation of Chinese Medicinal Materials in Southwest,Yunnan Agricultural University,Kunming 650201,China;College of Agriculture and Biotechnology,Yunnan Agricultural University,Kunming 650201,China)

机构地区:[1]云南农业大学,西南中药材种质创新与利用国家地方联合工程研究中心,云南昆明650201 [2]云南农业大学农学与生物技术学院,云南昆明650201

出  处:《云南农业大学学报:自然科学版》2019年第1期116-123,共8页Journal of Yunnan Agricultural University

基  金:国家自然基金项目(U1402262);云南省重大科技计划项目(2016ZF001);云南省科技计划项目(2012CG024)

摘  要:【目的】为更加精确探究外源物质赤霉素(GA3)、6-苄基腺嘌呤(6-BA)、脱落酸(ABA)、茉莉酸甲酯(MeJA)、水杨酸(SA)、萘乙酸(NAA)、乙烯(ETH)、褪黑素(MT)、亚精胺(Spd)、一氧化氮供体硝普钠(SNP)和过氧化氢(H2O2)对三七[Panax notoginseng(Burk.)F.H.Chen]叶皂苷含量的影响。【方法】开发超高效液相色谱—四极杆串联飞行时间质谱(UPLC-MS)结合高效液相色谱(HPLC)的新方法,测定三七叶中7种皂苷单体含量和生长指标。【结果】(1)建立的HPLC法可在52 min内准确测出三七叶中人参皂苷Rb1、Rc、Rb2和Rb3、三七皂苷Fa、Fe及七叶胆苷XVII等7种皂苷单体含量;(2)MeJA显著提高三七叶人参皂苷Rb1、Rc、Rb2、Rb3、三七皂苷Fa、Fe及七叶胆苷XVII的含量和总含量,且对三七生长无显著影响;而NAA、6-BA、H2O2、SNP、Spd和ABA显著提高三七皂苷Fe含量,ABA显著提高七叶胆苷XVII含量,但总含量均显著减少,且对三七生长有显著影响。【结论】建立了三七叶皂苷单体快速简便、稳定可靠的新检测方法,基于该方法明确了外源物质MeJA能大幅诱导三七叶皂苷单体的积累。[Purpose]We aimed to explore the effect of exogenous substances,including gibberellin(GA3),6-benzyladenine(6-BA),abscisic acid(ABA),methyl jasmonate(MeJA),salicylic acid(SA),naphthylacetate(NAA),ethylene(ETH),melatonin(MT),spermidine(Spd),sodium nitroprusside(SNP)and hydrogen peroxide(H2O2)on saponin contents of Panax notoginseng leaves.[Method]We developed a new method of ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-MS)together with high performance liquid chromatography(HPLC),and determined 7 saponin monomer contents and growth indexes.[Results](1)The established HPLC method could accurately measure 7 saponin monomer contents,including ginsenoside Rb1,Rc,Rb2,Rb3,notoginsenoside Fa,Fe and gypenoside XVII in P.notoginsen leaves within 52 minutes within a single run;(2)MeJA treatment significantly increased the contents of 7 monomer saponins,including ginsenoside Rb1,Rc,Rb2,Rb3,notoginsenoside Fa,Fe and gypenoside XVII in leaves,and had no effect on P.notoginseng growth.Other exogenous substances,including NAA,6-BA,H2O2,SNP and Spd significantly increased the content of notoginsenoside Fa,while ABA significantly improved the content of gypenoside XVII,but all of them significantly decreased the total saponin contents,and had effect on P.notoginseng growth.[Conclusion]A rapid and simple,stable and reliable detection method for saponin monomer of P.notoginseng leaves was established.Based on this method,it is clear that exogenous substance MeJA could significantly induce saponin monomer contents of P.notoginseng leaves.

关 键 词:三七叶 皂苷含量 外源物质 高效液相色谱 

分 类 号:S567.236.01[农业科学—中草药栽培;农业科学—作物学]

 

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