UPLC比较金银花、忍冬藤和忍冬叶中11种活性成分的含量
|
摘要
建立同时检测金银花、忍冬藤和忍冬叶中新绿原酸、绿原酸、咖啡酸、隐绿原酸、洋蓟素、异绿原酸A、异绿原酸B、异绿原酸C、芦丁、木樨草苷和马钱苷11种活性成分含量测定的UPLC分析方法,并比较金银花、忍冬藤、忍冬叶在酚酸类、黄酮类和环烯醚萜苷类成分含量上的差异。采用ACQUITY UPLC BEH C18色谱柱(2.1 mm×100 mm,1.7μm),乙腈-0.1%磷酸溶液为流动相,梯度洗脱,流速为0.3 mL·min-1;柱温30℃;样品室温度8℃;检测波长:新绿原酸、绿原酸、咖啡酸、隐绿原酸、洋蓟素、异绿原酸A、异绿原酸B和异绿原酸C为326 nm,芦丁和木樨草苷为352 nm,马钱苷为238 nm;进样量1μL。11种成分分离良好,各成分均有较宽的线性范围和良好的线性关系(r≥0.999 6),平均回收率(n=9)分别为98.96%,100.7%,97.24%,97.06%,99.53%,96.78%,98.12%,95.20%,95.12%,100.2%,98.61%;RSD分别为2.5%,1.4%,1.9%,2.1%,1.7%,1.9%,1.6%,2.0%,1.4%,2.2%,2.0%。基于含量测定结果,采用聚类分析和主成分分析等化学计量学方法对金银花、忍冬藤和忍冬叶药材进行了比较。结果表明金银花与忍冬叶化学成分较为相似,而与忍冬藤间存在差异,所建立的多组分定量分析方法可为金银花、忍冬藤、忍冬叶的质量控制提供方法参考。
This present study aims to establish a UPLC method for simultaneously determining eleven components such as new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,rutin,hibisin and loganin in Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica and comparing the differences in the contents of phenolic acids,flavonoids and iridoid glycosides of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The method was carried out on an ACQUITY UPLC BEH C18column(2.1 mm×100 mm,1.7 μm) by a gradient elution using acetonitrile and 0.1% phosphoric acid.The flow rate was 0.3 mL·min-1.The column temperature was maintained at 30 ℃.The sample room temperature was 8 ℃.The wavelength was set at 326 nm for new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C,352 nm for rutin and lignin,and 238 nm for loganin.The injection volume was 1 μL.The eleven components has good resolution and was separated to baseline.Each component had a wide linear range and a good linear relationship(r≥0.999 6),the average recovery rate(n=9) was 98.96%,100.7%,97.24%,97.06%,99.53%,96.78%,98.12%,95.20%,95.12%,100.2%,98.61%and with RSD was 2.5%,1.4%,1.9%,2.1%,1.7%,1.9%,1.6%,2.0%,1.4%,2.2%,2.0%,respectively.Based on the results of the content determination,the chemometric methods such as cluster analysis and principal component analysis were used to compare the Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The results showed that Lonicerae Japonicae Flos and leaves of Lonicera japonica were similar in the chemical constituents,but both showed chemical constituents difference compored to Lonicerae Japonicae Caulis.The established multi-component quantitative analysis method can provide a reference for the quality control of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.
This present study aims to establish a UPLC method for simultaneously determining eleven components such as new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B,isochlorogenic acid C,rutin,hibisin and loganin in Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica and comparing the differences in the contents of phenolic acids,flavonoids and iridoid glycosides of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The method was carried out on an ACQUITY UPLC BEH C18column(2.1 mm×100 mm,1.7 μm) by a gradient elution using acetonitrile and 0.1% phosphoric acid.The flow rate was 0.3 mL·min-1.The column temperature was maintained at 30 ℃.The sample room temperature was 8 ℃.The wavelength was set at 326 nm for new chlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,artichoke,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C,352 nm for rutin and lignin,and 238 nm for loganin.The injection volume was 1 μL.The eleven components has good resolution and was separated to baseline.Each component had a wide linear range and a good linear relationship(r≥0.999 6),the average recovery rate(n=9) was 98.96%,100.7%,97.24%,97.06%,99.53%,96.78%,98.12%,95.20%,95.12%,100.2%,98.61%and with RSD was 2.5%,1.4%,1.9%,2.1%,1.7%,1.9%,1.6%,2.0%,1.4%,2.2%,2.0%,respectively.Based on the results of the content determination,the chemometric methods such as cluster analysis and principal component analysis were used to compare the Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.The results showed that Lonicerae Japonicae Flos and leaves of Lonicera japonica were similar in the chemical constituents,but both showed chemical constituents difference compored to Lonicerae Japonicae Caulis.The established multi-component quantitative analysis method can provide a reference for the quality control of Lonicerae Japonicae Flos,Lonicerae Japonicae Caulis and leaves of Lonicera japonica.
引文
[1]赵金凤,周凤琴,郭庆梅,等.忍冬叶研究进展[J].中国药房,2010,21(39):3738.
[2]赵媛媛,杨倩茹,郝江波,等.金银花与忍冬藤及叶药理作用差异的研究进展[J].中国中药杂志,2016,41(13):2422.
[3]刘伟,杨帆,刘峰,等.忍冬叶活性成分周期变化及其代谢关系研究[J].中国中药杂志,2017,42(7):1292.
[4]张永欣,张启伟,李春,等.忍冬叶中抗氧化化学成分研究[J].中国中药杂志,2015,40(12):2372.
[5]马俊利,李宁,李铣.忍冬叶中咖啡酰奎宁酸类化学成分[J].中国中药杂志,2009,34(18):2346.
[6]耿姝,陈勇,金叶,等.基于近红外技术的金银花药材多指标成分快速检测[J].中国现代应用药学,2016,33(2):144.
[7]朱金花,王悠悠,庆伟霞,等.HPLC法同时测定忍冬叶中9种活性成分的研究[J].化学研究,2017,28(1):70.
[8]耿姝,陈勇,金叶,等.基于近红外技术的金银花药材多指标成分快速检测[J].中国现代应用药学,2016,33(2):144.
[9]苏静华,张超,孙磊,等.指纹图谱结合化学计量学用于香橼品种鉴别的可行性分析[J].中国中药杂志,2015,40(12):2318.
[10]吕露阳,张吉仲,张志锋,等.木香川木香药材UPLC特征指纹图谱建立及鉴别研究[J].中国中药杂志,2014,39(14):2699.
[11]周亚楠,孙菡,白洁,等.UHPLC特征图谱结合多组分含量评价首乌藤饮片的质量[J].中国现代应用药学,2018,35(8):1203.
[12]付绍智,王婷婷,高文远,等.基于主成分分析的不同初加工方法大黄的蒽醌及酚酸类成分比较研究[J].中国中药杂志,2014,39(5):833.
[13]张黄琴,严辉,钱大玮,等.不同产地百合药材中8种活性成分的分析与评价[J].中国中药杂志,2017,42(2):311.
[14]蔡晓洋,张思荻,曾俊,等.基于主成分分析和聚类分析的栀子种质资源评价[J].中国实验方剂学杂志,2017,23(14):30.
[2]赵媛媛,杨倩茹,郝江波,等.金银花与忍冬藤及叶药理作用差异的研究进展[J].中国中药杂志,2016,41(13):2422.
[3]刘伟,杨帆,刘峰,等.忍冬叶活性成分周期变化及其代谢关系研究[J].中国中药杂志,2017,42(7):1292.
[4]张永欣,张启伟,李春,等.忍冬叶中抗氧化化学成分研究[J].中国中药杂志,2015,40(12):2372.
[5]马俊利,李宁,李铣.忍冬叶中咖啡酰奎宁酸类化学成分[J].中国中药杂志,2009,34(18):2346.
[6]耿姝,陈勇,金叶,等.基于近红外技术的金银花药材多指标成分快速检测[J].中国现代应用药学,2016,33(2):144.
[7]朱金花,王悠悠,庆伟霞,等.HPLC法同时测定忍冬叶中9种活性成分的研究[J].化学研究,2017,28(1):70.
[8]耿姝,陈勇,金叶,等.基于近红外技术的金银花药材多指标成分快速检测[J].中国现代应用药学,2016,33(2):144.
[9]苏静华,张超,孙磊,等.指纹图谱结合化学计量学用于香橼品种鉴别的可行性分析[J].中国中药杂志,2015,40(12):2318.
[10]吕露阳,张吉仲,张志锋,等.木香川木香药材UPLC特征指纹图谱建立及鉴别研究[J].中国中药杂志,2014,39(14):2699.
[11]周亚楠,孙菡,白洁,等.UHPLC特征图谱结合多组分含量评价首乌藤饮片的质量[J].中国现代应用药学,2018,35(8):1203.
[12]付绍智,王婷婷,高文远,等.基于主成分分析的不同初加工方法大黄的蒽醌及酚酸类成分比较研究[J].中国中药杂志,2014,39(5):833.
[13]张黄琴,严辉,钱大玮,等.不同产地百合药材中8种活性成分的分析与评价[J].中国中药杂志,2017,42(2):311.
[14]蔡晓洋,张思荻,曾俊,等.基于主成分分析和聚类分析的栀子种质资源评价[J].中国实验方剂学杂志,2017,23(14):30.