指纹图谱结合化学计量法对葛根抗氧化活性部位的药效物质筛选
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篇名: 指纹图谱结合化学计量法对葛根抗氧化活性部位的药效物质筛选
TITLE: Effective Component Screening in Antioxidant Active Fraction of Pueraria lobata by Fingerprint Combined with Chemometrics
摘要: 目的:筛选葛根抗氧化活性部位的药效物质。方法:制备20批葛根抗氧化活性部位样品(S1~S20)。采用高效液相色谱法(HPLC)法测定,色谱柱为SepaxBio-C18,柱温为25℃,检测波长为250nm,流动相为甲醇-水(梯度洗脱),流速为0.6mL/min。采用《中药色谱指纹图谱相似度评价系统》(2012版)建立20批葛根抗氧化活性部位的HPLC指纹图谱并进行共有峰指认。结合聚类分析法、主成分分析(PCA)法和正交偏最小二乘法判别分析(OPLS-DA)法,筛选葛根抗氧化活性部位的药效物质。结果:20批葛根抗氧化活性部位HPLC指纹图谱共有18个共有峰,相似度均大于0.99。共指认了8个共有峰,分别为3′-羟基葛根素(峰2)、葛根素(峰3)、3′-甲氧基葛根素(峰4)、大豆苷(峰5)、染料木苷(峰7)、芒柄花苷(峰11)、大豆苷元(峰13)、染料木素(峰16);经聚类分析、PCA法分析发现,样品S1、S3、S4、S6、S8、S18、S19聚为一类,样品S2、S5、S7、S9~S17、S20聚为一类,对主成分1影响较大的有峰2、峰3、峰10、峰11、峰13,对主成分2影响较大有峰8、峰9;经OPLS-DA分析发现,峰4、峰3、峰2、峰16、峰13、峰11对葛根抗氧化活性部位质量的影响较大。结论:本研究建立了葛根抗氧化活性部位的HPLC指纹图谱,指认了8个成分,其中葛根素、3'-羟基葛根素、大豆苷元和芒柄花苷可能为葛根抗氧化作用的药效物质基础。
ABSTRACT: OBJECTIVE:To screen the effective compo nent in antioxi dant active fraction of Pueraria lobata . METHODS :The antioxidant active fraction sample (S1-S20) of 20 batches of P. lobata were prepared. HPLC method was adopted. The determination was performed on SepaxBio-C 18 column with mobile phase consisted of methanol-water (gradient elution )at the flow rate of 0.6 mL/min. The column temperature was set at 25 ℃,and detection wavelength was set at 250 nm. HPLC fingerprints of 20 batches of P. lobata were established by the Similarity Evaluation System of TCM Chromatographic Fingerprints (2012 edition),and common peaks were identified. Cluster analysis ,principal component analysis (PCA)and orthogonal partial least squares discriminant analysis (OPLS-DA)were used to screen the effective components in antioxidant active fraction of P. lobata . RESULTS:There were 18 common peaks in HPLC fingerprints of 20 batches of antioxidant active fraction in P. lobata ,and the similarity was more than 0.99. Eight common peaks were identified ,which were 3′-hydroxypuerarin(peak 2),puerarin(peak 3), 3′-methoxypuerarin(peak 4),daidzein(peak 5),genistein(peak 7),formononetin(peak 11),daidzein(peak 13)and genistein (peak 16). The results of cluster analysis and PCA analysis showed that samples S 1,S3,S4,S6,S8,S18 and S 19 were clustered into one category ,and samples S 2,S5,S7,S9-S17 and S 20 were clustered into one category ;peak 2,peak 3,peak 10,peak 11 and peak 13 had great influence on principal component 1;peak 8 and peak 9 had great influence on principal component 2. OPLS-DA analysis showed that peak 4,peak 3,peak 2,peak 16,peak 13 and peak 11 had great influence on the quality of antioxidant active fraction of P. lobata . CONCLUSIONS : HPLC fingerprint for active fraction of P. lobata is established in the study and 8 components are identified ;among them , com puerarin,3′-hydroxypuerarin,daidzein and formononetin maybe the material basis of antioxidant fraction of P. lobata .
期刊: 2021年第32卷第07期
作者: 庞会娜,范琳,肖凤琴,于倩,王海东,沈颖欣,韩荣欣,严铭铭,邵帅
AUTHORS: PANG Huina,FAN Lin,XIAO Fengqin,YU Qian,WANG Haidong,SHEN Yingxin,HAN Rongxin,YAN Mingming,SHAO Shuai
关键字: 葛根;抗氧化活性部位;高效液相色谱法;指纹图谱;化学计量法
KEYWORDS: Pueraria lobata ;Antioxidant fraction ;HPLC;
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