一测多评法测定罗布麻叶中6个黄酮类成分的含量
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摘要
目的:建立罗布麻叶中6个活性成分(芦丁、金丝桃苷、异槲皮苷、紫云英苷、槲皮素和山柰酚)同时测定的一测多评含量测定方法。方法:采用Waters XBridge C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.2%磷酸溶液为流动相,梯度洗脱,流速0.8 mL·min~(-1),检测波长256 nm,柱温35℃。以金丝桃苷为参照物建立与其他成分的相对校正因子,并计算其含量,实现一测多评。同时采用外标法测定该6个成分的含量,并比较二者的差异,以验证一测多评法的准确性和可行性。结果:金丝桃苷、芦丁、异槲皮苷、紫云英苷、槲皮素、山柰酚线性范围分别为11.36～284、0.416～10.4、9.2～230、4.08～102、0.812～20.3、0.272～6.81μg·mL~(~(-1)),金丝桃苷与芦丁、异槲皮苷、紫云英苷、槲皮素、山柰酚的相对校正因子值分别为1.378、0.974、1.197、0.867、1.114,且在不同实验条件下重复性良好(相对校正因子的RSD<3.0%),并与常规外标一点法比较,20批罗布麻叶中芦丁、金丝桃苷、异槲皮苷、紫云英苷、槲皮素、山柰酚的含量测定结果分别为0.066 57～0.178 4、2.766～3.576、2.794～3.334、0.271 9-0.668 5、0.078 65～0.256 5、0.018 72～0.106 2 mg·g~(-1),6个黄酮类成分的含量之和范围为6.343～7.732 mg·g~(-1)。一测多评法计算值与实测值间均无显著性差异(RSD<2.0%)。结论:以金丝桃苷建立的5个黄酮类成分相对校正因子可用于罗布麻叶的定量分析及质量评价。
Objective:To develop a method of quantitative analysis of multi-components by single marker(QAMS)for simultaneous determination of six bioactive constituents(rutin,hyperoside,isoquercitrin,astragalin,quercetin and kaempferol)in Apocyni Veneti Folium.Methods:A Waters XBridge C_(18) column(250 mm × 4.6 mm,5 μm)was used for the separation,with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 0.8 mL·min~(-1).The detection wavelength was set at 256 nm.The column temperature was 35℃.Hyperoside was selected as the internal reference substance,the relative correction factors(RCFs)between hyperoside and the other five flavanoids were established,and the contents of these six constituents in samples were calculated to realize QAMS.At the same time,the external standard method was used to quantify contents of the six constituents,the two methods were compared to validate the accuracy and feasibility of the new method.Results:The linear ranges of hyperoside,rutin,isoquercetin,astragaloside,quercetin and kaempferol were 11.36-284,0.416~(-1)0.4,9.2-230,4.08~(-1)02,0.812-20.3 and 0.272-6.8 μg·mL~(-1),respectively.RCFs of rutin,isoquercitrin,astragalin,quercetin and kaempferol with reference to hyperoside were 1.378,0.974,1.197,0.867 and 1.114,respectively,and repeatability was good in different experimental conditions(RSDs of RCFs were less than 3.0%).The contents of rutin,hyperoside,isoquercitrin,astragaloside,quercetin and kaempferol in 20 batches of Apocyni Veneti Folium were 0.066 57-0.178 4,2.766-3.576,2.794-3.334,0.271 9-0.668 5,0.078 65-0.256 5 and 0.018 72-0.106 2 mg·g~(-1),respectively.The range of total content of six flavonoids was 6.343-7.733 mg·g~(-1).No significant differences was found in the quantitative results of the six flavonoids in 20 batches of Apocyni Veneti Folium determined by external standard method and QAMS(RSD<2.0%).Conclusion:RCFs of 5 flavones with reference to hyperoside can be used for assay and quality evaluation of Apocyni Veneti Folium.
Objective:To develop a method of quantitative analysis of multi-components by single marker(QAMS)for simultaneous determination of six bioactive constituents(rutin,hyperoside,isoquercitrin,astragalin,quercetin and kaempferol)in Apocyni Veneti Folium.Methods:A Waters XBridge C_(18) column(250 mm × 4.6 mm,5 μm)was used for the separation,with gradient elution of acetonitrile and 0.2% phosphoric acid as the mobile phase at a flow rate of 0.8 mL·min~(-1).The detection wavelength was set at 256 nm.The column temperature was 35℃.Hyperoside was selected as the internal reference substance,the relative correction factors(RCFs)between hyperoside and the other five flavanoids were established,and the contents of these six constituents in samples were calculated to realize QAMS.At the same time,the external standard method was used to quantify contents of the six constituents,the two methods were compared to validate the accuracy and feasibility of the new method.Results:The linear ranges of hyperoside,rutin,isoquercetin,astragaloside,quercetin and kaempferol were 11.36-284,0.416~(-1)0.4,9.2-230,4.08~(-1)02,0.812-20.3 and 0.272-6.8 μg·mL~(-1),respectively.RCFs of rutin,isoquercitrin,astragalin,quercetin and kaempferol with reference to hyperoside were 1.378,0.974,1.197,0.867 and 1.114,respectively,and repeatability was good in different experimental conditions(RSDs of RCFs were less than 3.0%).The contents of rutin,hyperoside,isoquercitrin,astragaloside,quercetin and kaempferol in 20 batches of Apocyni Veneti Folium were 0.066 57-0.178 4,2.766-3.576,2.794-3.334,0.271 9-0.668 5,0.078 65-0.256 5 and 0.018 72-0.106 2 mg·g~(-1),respectively.The range of total content of six flavonoids was 6.343-7.733 mg·g~(-1).No significant differences was found in the quantitative results of the six flavonoids in 20 batches of Apocyni Veneti Folium determined by external standard method and QAMS(RSD<2.0%).Conclusion:RCFs of 5 flavones with reference to hyperoside can be used for assay and quality evaluation of Apocyni Veneti Folium.
引文
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[2]陆林玲,钱大玮,郭建明,等.一测多评法测定黄蜀葵花中7个黄酮类成分[J].药物分析杂志,2013,33(12):2082LU LL,QIAN DW,GUO JM,et al. A quantitative method using one marker for simultaneous assay of seven flavonoids in the flowers of Abelmoschus manihot.[J].Chin J Pharm Anal,2013,33(12):2082
[3]万青,涂楚月,熊慧,等.一测多评法测定藏药金腰草中4种黄酮类化学成分的含量[J].药物分析杂志,2016,36(6):1053WAN Q,TU CY,XIONG H,et al. Determination of four flavones in Chryso.splenium nudicaule Bunge by QAMS method[J].Chin J Pharm Anal,2016,36(6):1053
[4]陈建维,刘圆,刘晟楠,等.一测多评法测定枳实中4种黄酮类成分[J].中草药,2015,46(9):1374CHEN JW,LIU Y,LIU SN,et al. Determination of flavonoids in Aurantii Immaturus Fructus by multi-components with single marker[J].Chin Tradit Herb Drugs,2015,46(9):1374
[5]汪丽君,刘淑敏,毛春芹,等.一测多评法同时测定五味子中8个木脂素类成分的含量[J].药物分析杂志,2015,35(7):1191WANG LJ,LIU SM,MAO CQ,et al. Simultaneous determination of8 lignans in Schisandra chinensis by quantitative analysis of multicomponents by single marker[J].Chin J Pharm Anal,2015,35(7):1191
[6]梅国荣,刘飞,王福,等.一测多评法测定决明子中橙黄决明素、大黄素、大黄酚和大黄素甲醚[J].中草药,2016,47(8):1392MEI GR,LIU F,WANG F,et al. Determination of constituents of aurantio-obtusin,emodin,rhubarb,and physcion in Cassiae Semen by multi-components by single mark[J].Chin Tradit Herb Drugs,2016,47(8):1392
[7]钱正明,孙敏甜,艾中,等.一测多评法测定冬虫夏草中3种核昔的含量[J].中国药学杂志,2015,50(15):1297QIAN ZM,SUN MT,AI Z,et al. Simultaneous determination of three nucleosides in Cordyceps sinensis by QAMS[J].Chin Pharm J,2015,50(15):1297
[8]中华人民共和国药典2015年版.一部[S].2015:303,211ChP 2015.Vol I[S].2015:303,211
[9]侯晋军,韩利文,杨官娥,等.罗布麻叶化学成分和药理活性研究进展[J].中草药,2006,37(10):附7HOU JJ,HAN LW,YANG GE,et al. Advances on chemical constituents and pharmacological activities of Apocynm venetum L.[J].Chin Tradit Herb Drugs,2006,37(10):s7
[10]KUO CS,WAN CY,GONG CL,et al. Apocynum venetum leaf aqueous extract inhibits voltage-gated sodium channels of mouse neuroblastoma N2A cells[J].J Ethnopharmacol,2011,136(1):149
[11]XIE WY,ZHANG XY,WANG T,et al. Botany,traditional uses,phytochemistry and pharmacology of Apocyum venetum L.(Luobuma):a review[J].J Ethnopharmacol,2012,141:1
[12]XIONG QB,FAN WZ,TEZUKA Y,et al.Hepatoprotective effect of Apocynum venetum and its active constituents[J].Planta Med,2000,66(2):127
[13]林萍,易宏伟,张斐.金丝桃苷药理作用研究进展[J].中国现代中药,2012,14(10):23LIN P,YI HW,ZHANG F.Advances on pharmacological effects o hyperin[J].Mod Chin Med,2012,14(10):23
[14]李锦松,陈剑鸿,孟民杰.金丝桃苷药理作用及其作用机制的研究进展[J].广东药学院学报,2015,31(2):269LI JS,CHEN JH,MENG MJ.Progress on the pharmacologica effects and mechanism of hyperoside[J].J Guangdong Pharm Univ,2015,31(2):269
[15]郑荣,郭伟英.HPCE-DAD同时测定关东丁香叶中芦丁、紫云英苷、异槲皮苷[J].中成药,2013,35(11):2457ZHENG R,GUO WY.Simultaneous determination of rutin astragalin and isoquercetin in the leaves of Syringa veutina Kom.by HPCE-DAD[J]. Chin Tradit Pat Med,2013,35(11):2457
[16]臧志和,曹丽萍,钟铃.芦丁药理作用及制剂的研究进展[J].医药导报,2007,26(7):758ZANG ZH,CAO LP,ZHONG L.Research progress on pharmacological effects and preparations of rutin[J].Her Med,2007,26(7):758
[17]周浓,杨勤,杨敏,等.IPLC法同时测定川楝子中芦丁、异槲皮苷和槲皮素的含量[J].药物分析杂志,2013,33(2):225ZHOU N,YANG Q,YANG M,et al.Simultaneous determination of rutin,isoquercitrin and quercetin contents in Fructus Toosendan by HPLC[J].Chin J Pharm Anal,2013,33(2):225
[18]李玉英,赵淑娟,白崇智,等.苦荞异槲皮苷对人胃癌细胞SGC-7901增殖及凋亡的影响[J].食品科学,2014,35(3):193LI YY,ZHAO SJ,BAI CZ,et al. Effect of isoquercetin fron Fagopyrum tataricum on the proliferation and apoptosis of human gastric carcinoma cell line SGC-7901[J].J Food Sci,2014,35(3):193
[19]杨豪伟,朱盼盼,张春凤,等.紫云英苷对小鼠成骨细胞MC3T3-E1增殖与分化的影响[J].中医药学报,2013,41(4):17YANG HW,ZHU PP,ZHANG CF,et al. Influence of astragalin for proliferation and differentiation of mice's osteoblast MC3T3-E1[J].J Chin Med Pharmacol,2013,41(4):17
[20]汤喜兰,刘建勋,李澎,等.山柰酚和槲皮素对乳鼠心肌缺氧复氧及过氧化损伤的保护作用[J].中药药理与临床,2012,28(1):56.TANG XL,LIU JX,LI P,et al. Protective effects of kaempferol and quercetin on hypoxia/reoxygenation and peroxidation injury in neonatal cardiomyocytes[J].Pharmocol Clin Chin Mater Med,2012,28(1):56
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