不同产地葛根饮片的UPLC指纹图谱结合多成分含量测定研究
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摘要
建立不同产地葛根饮片的UPLC指纹图谱和同时测定6种异黄酮类化合物含量的UPLC分析方法。采用Waters ACQUITY UPLC BEH C_(18)色谱柱(2. 1 mm×50 mm,1. 7μm),流动相乙腈-0. 05%甲酸,梯度洗脱;检测波长250 nm;流速0. 2 mL·min~(-1);柱温30℃;进样量2μL;采用中药指纹图谱相似度评价系统进行相似度评价,并使用Simca-P软件中的主成分分析(PCA)与偏最小二乘法判别(OPLS-DA)寻找3个产地的差异成分。29批样品相似度均在0. 90以上,表明样品一致性良好,经PCA与OPLS-DA分析,葛根聚集为3组,并发现了葛根芹菜糖苷、大豆苷、大豆苷元等6个差异成分。对其中3'-羟基葛根素、葛根素、3'-甲氧基葛根素、葛根芹菜糖苷、大豆苷、大豆苷元6种异黄酮的含量测定结果表明,同一成分在各产地葛根饮片中的含量差异明显,不同成分的波动范围不同;各产地6种异黄酮质量分数为安徽11. 21%>河南10. 97%>陕西9. 38%。构建的不同产地葛根饮片UPLC指纹图谱结合多成分含量测定方法为完善葛根饮片的质量控制和品质评价提供了更全面的参考。
To establish ultra performance liquid chromatography( UPLC) fingerprint of Puerariae Lobatae Radix from different habitats and simultaneously determine the contents of six isoflavonoids. The UPLC fingerprint analysis and content determination were performed on a Waters ACQUITY UPLC BEH C_(18)( 2. 1 mm×50 mm,1. 7 μm) chromatographic column,with acetonitrile-0. 05% formic acid as mobile phase for gradient elution. The detection wavelength was set at 250 nm; the flow rate was 0. 2 mL·min~(-1); the column temperature was 30 ℃ and the injection volume was 2 μL. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine( TCM) was adopted; principal component analysis( PCA) and discriminant analysis by partial least square method( PLS-DA) in Simca-P software were used to identify the differential components in samples from three habitats. The similarity was over 0. 90 in 29 batches of samples,indicating good consistency of the samples. The samples were clustered into 3 categories by PCA and PLS-DA,and six differential components such as puerarin apioside,daidzin,and isoflavoues aglycone were found. The determination results of 6 isoflavones,including 3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,puerarin apioside,daidzin,and isoflavoues aglycone,showed that the content of the same component and the fluctuation range between different components were all different among different habitats. The total content of 6 isoflavones from different regions was Anhui 11. 21% >Henan 10. 97% >Shannxi 9. 38%. The establishment of UPLC fingerprint combined with simultaneous determination of 6 active components provides a more comprehensive reference for quality control and quality evaluation of Puerariae Lobatae Radix.
To establish ultra performance liquid chromatography( UPLC) fingerprint of Puerariae Lobatae Radix from different habitats and simultaneously determine the contents of six isoflavonoids. The UPLC fingerprint analysis and content determination were performed on a Waters ACQUITY UPLC BEH C_(18)( 2. 1 mm×50 mm,1. 7 μm) chromatographic column,with acetonitrile-0. 05% formic acid as mobile phase for gradient elution. The detection wavelength was set at 250 nm; the flow rate was 0. 2 mL·min~(-1); the column temperature was 30 ℃ and the injection volume was 2 μL. Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine( TCM) was adopted; principal component analysis( PCA) and discriminant analysis by partial least square method( PLS-DA) in Simca-P software were used to identify the differential components in samples from three habitats. The similarity was over 0. 90 in 29 batches of samples,indicating good consistency of the samples. The samples were clustered into 3 categories by PCA and PLS-DA,and six differential components such as puerarin apioside,daidzin,and isoflavoues aglycone were found. The determination results of 6 isoflavones,including 3'-hydroxy puerarin,puerarin,3'-methoxy puerarin,puerarin apioside,daidzin,and isoflavoues aglycone,showed that the content of the same component and the fluctuation range between different components were all different among different habitats. The total content of 6 isoflavones from different regions was Anhui 11. 21% >Henan 10. 97% >Shannxi 9. 38%. The establishment of UPLC fingerprint combined with simultaneous determination of 6 active components provides a more comprehensive reference for quality control and quality evaluation of Puerariae Lobatae Radix.
引文
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[2]时晓媞,贺志龙,顾健,等.基于5种葛根异黄酮组分综合评分法优选葛根提取工艺[J].现代中药研究与实践,2016(3):49.
[3]侯筱婷.葛根素对心脑血管疾病影响的研究进展[J].中西医结合心脑血管病杂志,2018,16(14):2002.
[4]刘倩,范颖,李新,等.黄芪葛根配伍调节糖尿病大鼠血糖血脂炎症的作用与机制[J].中国实验方剂学杂志,2018,24(11):81.
[5] Ge B,Zhang Z,Zhong Z. Radix Puerariae Lobatae(Gegen)suppresses the anticoagulation effect of warfarin:a pharmacokinetic and pharmacodynamics study[J]. Chin Med,2016,11(1):1.
[6]钟凌云,马冰洁,叶喜德,等.葛根主要药效成分止泻作用研究[J].世界科学技术——中医药现代化,2015,17(1):109.
[7] Liu M,Liu R,Wu H,et al. Radix Puerariae extracts ameliorate paraquat-induced pulmonary fibrosis by attenuating follistatinlike 1 and nuclear factor erythroid 2p45-related factor-2 signalling pathways through downregulation of miRNA-21 expression[J].BMC Complement Altern Med,2015,16(1):1.
[8]邹纯才,鄢海燕.我国中药色谱指纹图谱相似度评价方法30年(1988—2017年)研究进展与展望[J].中国中药杂志,2018,43(10):1969.
[9]刘东方,赵丽娜,李银峰,等.中药指纹图谱技术的研究进展及应用[J].中草药,2016,47(22):4085.
[10]郝敏,陆兔林,毛春芹,等. 3种温郁金根茎炮制品的UPLC指纹图谱与多成分含量测定研究[J].中国中药杂志,2018,43(11):2288.
[11]刘爱朋,郭利霄,薛紫鲸,等.基于指纹图谱和多组分含量测定的蒲公英药材质量控制研究[J].中国中药杂志,2018,43(18):3715.
[12]张帅,丛海林,于冰.超高效液相色谱的发展及在分析领域的应用[J].分析仪器,2017(6):16.
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