川芎饮片的HPLC指纹图谱建立、聚类分析及偏最小二乘判别分析
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摘要
目的:建立川芎饮片的高效液相色谱(HPLC)指纹图谱,并进行聚类分析和偏最小二乘判别(PLS-DA)分析。方法:采用HPLC法,色谱柱为Waters Symmetry C_(18),流动相为乙腈-0.5%醋酸水溶液(梯度洗脱),流速为1 mL/min,检测波长为254 nm,柱温为30℃,进样量为10μL。以藁本内酯为参照,绘制21批样品(S1～S21)的HPLC图谱;采用《中药色谱指纹图谱相似度评价系统》(2012 A版)进行相似度评价,确定共有峰;采用SPSS 19.0软件进行聚类分析,并结合PLS-DA分析区分样品。结果:21批样品的HPLC图谱有25个共有峰,并指认了其中9个共有峰;相似度为0.769～0.989,其中基地、传统药用部位样品(S1～S10)相似度均大于0.970。21批样品可聚为3类,S1～S10聚为一类,S15～S16、S19～S20聚为一类,其余聚为一类。PLS-DA分析确定了11个分类标志物,并指认了阿魏酸、阿魏酸松柏酯、正丁基苯酞、藁本内酯、洋川芎内酯A等5个色谱峰,这5个色谱峰可有效区分非市售、基地样品(S1～S10)与市售、非基地样品(S11～S21),与聚类分析结果一致。结论:所建指纹图谱、聚类分析及PLS-DA分析结果可为川芎饮片的质量评价提供参考。
OBJECTIVE:To establish HPLC fingerprints of Ligusticum chuanxiong decoction pieces,and to conduct cluster analysis and PLS-DA analysis. METHODS:HPLC method was adopted. The determination was performed on Waters Symmetry C_(18) column with mobile phase consisted of acetonitrile-0.5% acetic acid solution(gradient elution)at the flow rate of 1 mL/min. The detection wavelength was set at 254 nm,and the column temperature was 30 ℃. The sample size was 10 μL. Using ligustilide as control,HPLC chromatograms of 21 batches of samples(S1-S20)were determined. The similarity evaluation was conducted by using Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition) to determine common peak. Cluster analysis was conducted by using SPSS 19.0 software and PLS-DA was used to distinguish the samples. RESULTS:There were 25 common peaks in HPLC chromatograms for 21 batches of samples,and 9 common peaks were identified. The similarity of samples was between 0.768-0.989,and the similarity of base and traditional medicinal part samples(S1-S10)were more than 0.970. The 21 batches of samples were clustered into 3 categories,in which S1-S10 were category Ⅰother were category Ⅲ. By PLS-DA analysis,11 classification markers were identified as well as 5 chromatogram peaks were identified,such as ferulic acid,pine cyperyl ferulate,n-butyl phthalide,ligustilide,ligustilide A,which could be used to distinguish base and non-markted samples(S1-S10)from marketed and non-base samples(S11-S21),which were consistent with the results of cluster analysis. CONCLUSIONS: Established fingerprint, cluster analysis and PLS-DA analysis can provide reference for quality evaluation of L. chuanxiong decoction pieces.
OBJECTIVE:To establish HPLC fingerprints of Ligusticum chuanxiong decoction pieces,and to conduct cluster analysis and PLS-DA analysis. METHODS:HPLC method was adopted. The determination was performed on Waters Symmetry C_(18) column with mobile phase consisted of acetonitrile-0.5% acetic acid solution(gradient elution)at the flow rate of 1 mL/min. The detection wavelength was set at 254 nm,and the column temperature was 30 ℃. The sample size was 10 μL. Using ligustilide as control,HPLC chromatograms of 21 batches of samples(S1-S20)were determined. The similarity evaluation was conducted by using Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition) to determine common peak. Cluster analysis was conducted by using SPSS 19.0 software and PLS-DA was used to distinguish the samples. RESULTS:There were 25 common peaks in HPLC chromatograms for 21 batches of samples,and 9 common peaks were identified. The similarity of samples was between 0.768-0.989,and the similarity of base and traditional medicinal part samples(S1-S10)were more than 0.970. The 21 batches of samples were clustered into 3 categories,in which S1-S10 were category Ⅰother were category Ⅲ. By PLS-DA analysis,11 classification markers were identified as well as 5 chromatogram peaks were identified,such as ferulic acid,pine cyperyl ferulate,n-butyl phthalide,ligustilide,ligustilide A,which could be used to distinguish base and non-markted samples(S1-S10)from marketed and non-base samples(S11-S21),which were consistent with the results of cluster analysis. CONCLUSIONS: Established fingerprint, cluster analysis and PLS-DA analysis can provide reference for quality evaluation of L. chuanxiong decoction pieces.
引文
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[11]王娣,刘玉红,刘云华,等.川芎药材及饮片质量标准提升研究[J].亚太传统医药,2017,13(5):34-38.
[12]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):4085-4094.
[13]高微,陈明生,韦广辉,等.尖尾风药材的HPLC指纹图谱建立及聚类分析和主成分分析[J].中国药房,2018,29(16):2215-2219.
[14]黎理,曾祥燕,谢凤凤,等.竹节蓼药材的HPLC指纹图谱建立及聚类分析[J].中国药房,2018,29(12):1640-1643.
[15]马云桐,徐世军,廖海浪,等.HPLC同时测定川芎、当归SFE-CO2萃取物中7种成分含量及阿魏酸松柏酯稳定性考察[J].中国实验方剂学杂志,2017,23(9):24-29.
[2]梅超南,曾瑾,张了云,等.不同产地川芎对家兔血小板聚集、小鼠凝血功能及血瘀大鼠血液流变性的品质评价研究[J].中药药理与临床,2014,30(2):110-112.
[3]国家药典委员会.中华人民共和国药典:一部[S].2015年版.北京:中国医药科技出版社,2015:40-41.
[4]CHANG XL,JIANG ZY,MA YB,et al.Two new compounds from the roots of Ligusticum chuanxiong[J].JAsian Nat Prod Res,2009,11(9):805-810.
[5]CHEN ZJ,ZHANG C,GAO F,et al.A systematic review on the rhizome of Ligusticum chuanxiong Hort.(Chuanxiong)[J].Food Chem Toxicol,2018.DOI:10.1016/j.fct.2018.02.050.
[6]LI WX,TANG YP,CHEN YY,et al.Advances in the Chemical Analysis and Biological Activities of Chuanxiong[J].Molecules,2012,17(9):10614-10651.
[7]韩炜.川芎的化学成分与药理作用研究进展[J].中国现代中药,2017,19(9):1341-1349.
[8]张丽娟,刘继勇,姚翀,等.洋川芎内酯类化合物药理作用研究进展[J].中国药学杂志,2015,50(13):1081-1084.
[9]赵润英,郝伟,孟祥军,等.阿魏酸川芎嗪抗血栓形成作用及其对血小板中性粒细胞黏附的影响[J].中国医科大学学报,2012,41(10):900-903、925.
[10]梁乙川,郭换,刘素娟,等.一测多评法测定川芎中7种成分含量[J].中药材,2018,41(1):140-143.
[11]王娣,刘玉红,刘云华,等.川芎药材及饮片质量标准提升研究[J].亚太传统医药,2017,13(5):34-38.
[12]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):4085-4094.
[13]高微,陈明生,韦广辉,等.尖尾风药材的HPLC指纹图谱建立及聚类分析和主成分分析[J].中国药房,2018,29(16):2215-2219.
[14]黎理,曾祥燕,谢凤凤,等.竹节蓼药材的HPLC指纹图谱建立及聚类分析[J].中国药房,2018,29(12):1640-1643.
[15]马云桐,徐世军,廖海浪,等.HPLC同时测定川芎、当归SFE-CO2萃取物中7种成分含量及阿魏酸松柏酯稳定性考察[J].中国实验方剂学杂志,2017,23(9):24-29.
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