指纹图谱、多成分定量分析及化学模式识别分析相结合的心安胶囊质量评价
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篇名: 指纹图谱、多成分定量分析及化学模式识别分析相结合的心安胶囊质量评价
TITLE: Quality evaluation of Xin ’an capsules by combination of fingerprint ,multi-component quantitative analysis and chemical pattern recognition analysis
摘要: 目的 建立指纹图谱、多成分定量分析和化学模式识别分析相结合的心安胶囊质量评价方法。方法采用高效液相色谱(HPLC)法,结合《中药色谱指纹图谱相似度评价系统(2012版)》建立24批心安胶囊的指纹图谱,并进行相似度评价,确定共有峰;采用同一HPLC法测定心安胶囊中牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、金丝桃苷和异槲皮素的含量;以指纹图谱共有峰峰面积为变量,采用MetaboAnalyst5.0工具绘制聚类分析(CA)热图,采用SIMCA14.1软件进行主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)。结果24批心安胶囊共有12个共有峰,相似度均大于0.97;指认出绿原酸、牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、金丝桃苷和异槲皮素6个共有峰。牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、金丝桃苷、异槲皮素检测质量浓度的线性范围分别为2.36~151.35、9.15~585.20、1.20~76.50、0.68~43.20、0.44~27.90µg/mL(r均大于0.999);精密度、重复性、稳定性(24h)试验的RSD均小于2.00%;平均加样回收率分别为95.80%(RSD=0.96%,n=6)、102.10%(RSD=0.93%,n=6)、103.26%(RSD=1.28%,n=6)、103.89%(RSD=0.73%,n=6)、102.09%(RSD=1.79%,n=6)。5种成分的含量分别为0.9888~1.5591、4.3366~11.2201、0.0651~0.8305、0.0438~0.6925、0.0232~0.4272mg/粒。CA、PCA结果显示,24批样品可分成3类,S1~S15为一类,S16~S18为一类,S19~S24为一类;PLS-DA发现,6号峰对应成分和牡荆素葡萄糖苷(7号峰)、牡荆素鼠李糖苷(8号峰)、金丝桃苷(10号峰)、异槲皮素(11号峰)的变量重要性投影值均大于1。结论所建立的HPLC指纹图谱、多成分定量方法简单、可行,结合化学模式识别分析可用于心安胶囊的质量控制;牡荆素葡萄糖苷、牡荆素鼠李糖苷等可能是影响各批样品质量的差异标志物。
ABSTRACT: OBJE CTIVE To establish a method for quality evaluation of Xin ’an capsule by combining fingerprint , multi-component quantitative analysis and chemical pattern recognition analysis. METHODS High performance liquid chromatography(HPLC)combined with Similarity Evaluation System of TCM Chromatogram Fingerprint (2012 edition)were used to establish the fingerprints of 24 batches of Xin ’an capsules and evaluate the similarity. The common peaks were determined. The contents of glucosylvitexin ,rhamnosylvitexin,vitexin,hyperoside and isoquercetin in Xin ’an capsules were determined by the same HPLC method. Taking the common peak area of fingerprint as the variable ,MetaboAnalyst 5.0 tool was used to draw the cluster analysis (CA)heat map. SIMCA 14.1 software was used to perform principle component analysis (PCA)and partial least squares-discriminant analysis (PLS-DA). RESULTS Twelve common peaks were identified with the similarity greater than 0.97. Six common peaks were identified as chlorogenic acid ,glucosylvitexin,rhamnosylvitexin,vitexin,hyperoside and isoquercetin.The linear range of glucosylvitexin ,rhamnosylvitexin,vitexin, hyperoside and isoquercetin were 2.36-151.35,9.15-585.20, 1.20-76.50, 0.68-43.20, 0.44-27.90 µg/mL(all r>0.999).RSDs of precision ,repeatability and stability (24 h)tests were 163.com all less than 2.00% . The average recoveries were 95.80%(RSD=0.96% ,n=6),102.10% (RSD=0.93% ,n=6), 103.26%(RSD=1.28%,n=6),103.89%(RSD=0.73%,n=6) and 102.09%(RSD=1.79%,n=6),respectively. The contents of the five components were 0.988 8-1.559 1,4.336 6-11.220 1, 0.065 1-0.830 5,0.043 8-0.692 5 and 0.023 2-0.427 2 mg/grain,respectively. The results of CA and PCA showed that 24 batches of samples could be divided into three categories ,i.e. S 1-S15,S16-S18 and S 19-S24. PLS-DA showed that variable importance in projection values of the corresponding component of peak 6 and glucosylvitexin (peak 7),rhamnosylvitexin(peak 8),hyperoside (peak 10) and isoquercetin (peak 11) were greater than 1. CONCLUSIONS The established HPLC fingerprint and multi-component quantitative method are simple and feasible. Combined with chemical pattern recognition analysis ,it can be used for the quality control of Xin ’an capsules. Glucosylvitexin ,rhamnosylvitexin and other components may be differentital markers affecting the quality of each batch of samples.
期刊: 2022年第33卷第06期
作者: 李艳荣,杜义龙,沈莹,吴哲,王狄鑫,赵胜男,潘海峰
AUTHORS: LI Yanrong ,DU Yilong ,SHEN Ying,WU Zhe,WANG Dixin,ZHAO Shengnan ,PAN Haifeng
关键字: 心安胶囊;高效液相色谱法;指纹图谱;多成分定量分析;聚类分析;主成分分析;偏最小二乘法-判别分析;质量控制
KEYWORDS: Xin’an capsules ;high performance liquid chromatography ;fingerprint;multi-component quantitative analysis ;
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