一测多评法测定蓄中3个黄酮类成分的含量
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摘要
目的:以第四次全国中药资源普查实地采集的76批蓄药材为样本,研究建立蓄一测多评的含量测定方法,考察同一类型黄酮类化合物之间采用相对校正因子进行准确定量的可行性。方法:采用UPLC进行含量测定,色谱柱为Phenomenex Kinetex C_(18)(2.1 mm×100 mm,1.7μm);流动相为乙腈-0.5%磷酸水溶液,线性梯度洗脱;柱温45℃;流速0.2 m L·min~(-1);检测波长360 nm;以蓄为研究对象,以杨梅苷为参照物,测定杨梅苷、蓄苷与槲皮苷的相对校正因子,并利用该校正因子,计算蓄苷、槲皮苷的含量,实现一测多评;同时采用外标法测定蓄中杨梅苷、蓄苷及槲皮苷的含量,将其测定值与一测多评法计算值进行对比分析。结果:杨梅苷、蓄苷与槲皮苷分别在2.42～48.4 mg·L~(-1)(r=0.999 8),2.314～46.28mg·L~(-1)(r=0.999 9)和5.523～50.46 mg·L~(-1)(r=0.999 9)范围内线性关系良好,回收率分别为101.2%、98.9%和97.6%。试验中采用夹角余弦法统计分析2种方法测得的76批蓄药材中蓄苷和槲皮苷含量数据,夹角余弦值分别为0.999 8、0.999 9、0.999 8,2组数据无显著性差异。结论:本研究建立的以杨梅苷对照品为参照物,采用相对校正因子实现对蓄苷和槲皮苷的含量测定方法是准确的、可行的,一测多评法可以实现对同类型化合物进行准确定量。
Objective:Using 76 batches of samples collected from the 4 th national Chinese Materia Medica Resource Inventory,to establish a method for simultaneous determination of three flavonoids in Polygoni Avicularis Herba by quantitative analysis of multi-components with single marker(QAMS)strategy and investigate the feasibility of using correction factors between the identical types of flavonoids.Methods:UPLC analysis was performed on a Phenomenex Kinetex C_(18) column(2.1 mm×100 mm,1.7 μm).The mobile phase consisted of acetonitrile(solvent A)and 0.5% phosphoric acid water solution(solvent B)with linear gradient elution at a flow rate of 0.2 mL·min~(-1).The detection wavelength and column temperature were 360 nm and 45 ℃,respectively.The relative correction factors(RCFs)of curcumin to the other two ingredients were calculated.The method was evaluated by comparison of the quantitative results between external standard method and QAMS method.Results:Good linear relationships were observed for myricitrin,quercitrin and avicularin within the ranges of 2.42-48.4 mg·L~(-1)(r=0.999 8),2.314-46.28 mg·L~(-1)(r=0.999 9)and 5.523-50.46 mg·L~(-1)(r=0.999 9),respectively.The average recoveries(n=6)were 101.2% for myricitrin,98.9% for quercitrin,97.6% for avicularin,respectively.No obvious differences were found in the quantitative results of myricetrin,avicularin and quercitrin in 76 batches of Polygoni Avicularis Herba determined by the two methods,according to the angle cosine value,the angle cosine values were 0.999 8 for myricitrin,0.999 9 for quercitrin and 0.999 8 for avicularin.Conclusion:It is feasible and suitable to determine myricetrin,avicularin and quercitrin in Polygoni Avicularis Herba by QAMS,and this method can be used for quality evaluation of the identical types of flavonoids.
Objective:Using 76 batches of samples collected from the 4 th national Chinese Materia Medica Resource Inventory,to establish a method for simultaneous determination of three flavonoids in Polygoni Avicularis Herba by quantitative analysis of multi-components with single marker(QAMS)strategy and investigate the feasibility of using correction factors between the identical types of flavonoids.Methods:UPLC analysis was performed on a Phenomenex Kinetex C_(18) column(2.1 mm×100 mm,1.7 μm).The mobile phase consisted of acetonitrile(solvent A)and 0.5% phosphoric acid water solution(solvent B)with linear gradient elution at a flow rate of 0.2 mL·min~(-1).The detection wavelength and column temperature were 360 nm and 45 ℃,respectively.The relative correction factors(RCFs)of curcumin to the other two ingredients were calculated.The method was evaluated by comparison of the quantitative results between external standard method and QAMS method.Results:Good linear relationships were observed for myricitrin,quercitrin and avicularin within the ranges of 2.42-48.4 mg·L~(-1)(r=0.999 8),2.314-46.28 mg·L~(-1)(r=0.999 9)and 5.523-50.46 mg·L~(-1)(r=0.999 9),respectively.The average recoveries(n=6)were 101.2% for myricitrin,98.9% for quercitrin,97.6% for avicularin,respectively.No obvious differences were found in the quantitative results of myricetrin,avicularin and quercitrin in 76 batches of Polygoni Avicularis Herba determined by the two methods,according to the angle cosine value,the angle cosine values were 0.999 8 for myricitrin,0.999 9 for quercitrin and 0.999 8 for avicularin.Conclusion:It is feasible and suitable to determine myricetrin,avicularin and quercitrin in Polygoni Avicularis Herba by QAMS,and this method can be used for quality evaluation of the identical types of flavonoids.
引文
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[12]连赟芳,陈丹,蔡韦炜,等.一测多评法同时测定玳玳果黄酮滴丸中4个活性成分的含量[J].药物分析杂志,2015,35(6):974LIAN YF,CHEN D,CAI WW,et al.Determination of four active components in Daidaiguo Huangtong dropping pills by QAMS[J].Chin J Pharm Anal,2015,35(6):974
[13]李木子,王京辉,郭洪祝,等.HPLC法测定白术饮片中多种化学成分的含量[J].药物分析杂志,2017,37(9):1585LI MZ,WANG JH,GUO HZ,et,al.Simultaneous determination of multiple components in decoction pieces of Atractylodes macrocephala by HPLC[J].Chin J Pharm Anal,2017,37(9):1585
[14]杨海玲,吴丽丹,覃德杰,等.一测多评法同时测定广西姜黄饮片中3种姜黄素类成分的含量[J].药物分析杂志,2016 36(9):1571YANG HL,WU LD,QIN DJ,et,al.Simultaneous determination of three curcumin ingredients in Rhizoma Curcumae Longae from Guangxi Province by QAMS[J].Chin J Pharm Anal,2016,36(9):1571
[15]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 flvones in Chrysosplenium nudicaule Bunge by QAMS method[J].Chin JPharm Anal,2016,36(6):105
[16]王龙星,肖红斌,梁鑫淼.一种评价中药色谱指纹谱相似性的新方法:向量夹角法[J].药学学报,2002,37(9):713WANG LX,XIAO HB,LIANG XM.Vectorial angle method for evaluating the similarity between two chromatographic fingerprints of Chinese herb[J].Acta Pharm Sin,2002,37(9):713
[17]赵大鹏,侯元同.中国蓼族植物(蓼科)的地理分布[J].中国野生植物资源,2011,30(3):14ZHAO DP,HOU YT.The geographic distribution of the Ttribe Polygoneae(Polygonaceae)in China[J].Chin Wild Plant Resour,2011,30(3):14
[18]李景华,万玉梅,刘玉芹.扁蓄种群生命表的研究[J].吉林医药学院学报,2005,26(3):137LI JH,WAN YM,LIU YQ.Study on life table of Polygonum aviculare population[J].J Jilin Med Coll,2005,26(3):137
[2]陈晓虎,陈道峰.萹蓄化学成分研究[J].中国中药杂志,2004,29(9):918CHEN XH,CHEN DF.Studies on the chemical constituents of Polygonum aviculare L.[J].China J Chin Mater Med,2004,29(9):918
[3]于兆海,孙结.萹蓄的化学成分研究[J].中草药,2002,33(6):498YU ZH,SUN J.Studies on the chemical constituents of Polygonum aviculare L.[J].Chin Tradit Herb Drug,2002,33(6):498
[4]李曼曼,刘增辉,王海燕,等.萹蓄抑菌活性及化学成分研究[J].天然产物研究与开发,2014,26(4):526.LI MM,LIU ZH,WANG HY,et al.Studies on the antibacterial activities and chemical constituents of Polygonum aviculare L.[J].Nat Prod Res Dev,2014,26(4):526
[5]许福泉,刘红兵,罗建光,等.萹蓄化学成分及其归经药性初探[J].中国海洋大学学报,2010,40(3):101XU FQ,LIU HB,LUO JG,et al.Studies on the chemical constituents and meridian doctrine of Polygonum aviculare L.[J].Period Ocean Univ China,2010,40(3):101
[6]GRANICA S,CZERWI?SKA ME,?Y?Y?SKA-GRANICA B,et al.Antioxidant and anti-inflammatory flavonol glucuronides from Polygonum aviculare L.[J].Fitoterapia,2013,91:180
[7]AGUNG N,EON JK,JAE SC,et al.Simultaneous quantification and peroxynitrite-scavenging activities of flavonoids in Polygonum aviculare L.herb[J].J Pharm Biomed Anal,2014,89:93
[8]FU QR,LIU SJ,WANG H,et al.Simultaneous determination of eight flavonoids in Polygonum aviculare L.by RP-HPLC-UV[J].JChin Pharm Sci,2014,23(3):170
[9]陈娟,师彦平.HPLC测定萹蓄中槲皮素和山柰素的含量[J].中国中药杂志,2009,34(4):423CHEN J,SHI YP.Determination of quercetin and kaempferol in Polygonum aviculare L.by HPLC[J].China J Chin Mater Med,2009,34(4):423
[10]王智民,钱忠直,张啓伟,等.“一测多评”法建立的技术指南[J].中国中药杂志,2011,36(6):657WANG ZM,QIAN ZZ,ZHANG QW,et,al.Technical guide for QAMS[J].China J Chin Mater Med,2011,36(6):657
[11]马会利,郭庆梅,周凤琴.“一测多评法”在中药质量控制中应用研究进展[J].山东中医药大学学报,2014,38(3):282MA HL,GUO QM,ZHOU FQ.Multi-components quantitation by one maker new method applied research progress in quality control of traditional Chinese medicine[J].J Shandong Univ Tradit Chin Med,2014,38(3):282
[12]连赟芳,陈丹,蔡韦炜,等.一测多评法同时测定玳玳果黄酮滴丸中4个活性成分的含量[J].药物分析杂志,2015,35(6):974LIAN YF,CHEN D,CAI WW,et al.Determination of four active components in Daidaiguo Huangtong dropping pills by QAMS[J].Chin J Pharm Anal,2015,35(6):974
[13]李木子,王京辉,郭洪祝,等.HPLC法测定白术饮片中多种化学成分的含量[J].药物分析杂志,2017,37(9):1585LI MZ,WANG JH,GUO HZ,et,al.Simultaneous determination of multiple components in decoction pieces of Atractylodes macrocephala by HPLC[J].Chin J Pharm Anal,2017,37(9):1585
[14]杨海玲,吴丽丹,覃德杰,等.一测多评法同时测定广西姜黄饮片中3种姜黄素类成分的含量[J].药物分析杂志,2016 36(9):1571YANG HL,WU LD,QIN DJ,et,al.Simultaneous determination of three curcumin ingredients in Rhizoma Curcumae Longae from Guangxi Province by QAMS[J].Chin J Pharm Anal,2016,36(9):1571
[15]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 flvones in Chrysosplenium nudicaule Bunge by QAMS method[J].Chin JPharm Anal,2016,36(6):105
[16]王龙星,肖红斌,梁鑫淼.一种评价中药色谱指纹谱相似性的新方法:向量夹角法[J].药学学报,2002,37(9):713WANG LX,XIAO HB,LIANG XM.Vectorial angle method for evaluating the similarity between two chromatographic fingerprints of Chinese herb[J].Acta Pharm Sin,2002,37(9):713
[17]赵大鹏,侯元同.中国蓼族植物(蓼科)的地理分布[J].中国野生植物资源,2011,30(3):14ZHAO DP,HOU YT.The geographic distribution of the Ttribe Polygoneae(Polygonaceae)in China[J].Chin Wild Plant Resour,2011,30(3):14
[18]李景华,万玉梅,刘玉芹.扁蓄种群生命表的研究[J].吉林医药学院学报,2005,26(3):137LI JH,WAN YM,LIU YQ.Study on life table of Polygonum aviculare population[J].J Jilin Med Coll,2005,26(3):137