辣椒药材的HPLC指纹图谱建立及聚类分析和主成分分析
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摘要
目的:建立辣椒药材的高效液相色谱(HPLC)指纹图谱,并进行聚类分析和主成分分析。方法:采用HPLC法。色谱柱为Agilent C18,流动相为甲醇-0.2%磷酸水溶液(梯度洗脱),流速为1.0 mL/min,检测波长为235 nm,柱温为30℃,进样量为15μL。以辣椒素峰为参照,绘制15批药材样品的HPLC指纹图谱,采用《中药色谱指纹图谱相似度评价系统》(2004 A版)进行相似度评价,确定共有峰,并采用SPSS 19.0软件进行聚类分析和主成分分析。结果:15批药材样品的HPLC图谱相似度均在0.95以上;有12个共有峰,并指认了辣椒素峰;聚类分析结果显示,15批药材样品可聚为3类,S1、S3～S5、S7、S9～S13聚为一类,S2、S14、S15聚为一类,S6、S8聚为一类。经主成分分析,4个主成分因子的累积方差贡献率为94.093%,以S5药材样品的主成分因子综合得分最高、整体质量最好。结论:所建HPLC指纹图谱及聚类分析和主成分分析结果可为辣椒药材的质量控制提供参考。
OBJECTIVE:To establish HPLC fingerprint of Capsicum annuum,and to conduct cluster analysis and principal component analysis. METHODS:HPLC method was adopted. The determination was performed on Agilent C_(18) column with mobile phase consisted of methanol-0.2% phosphoric acid solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 235 nm,and column temperature was 30 ℃. The sample size was 15 μL. Using capsaicin peak as reference,HPLC fingerprints of 15 batches of C. annuum from different production areas were determined. The similarity evaluation ofcommon peaks was evaluated by using the TCM Chromatographic Fingerprint Similarity Evaluation System(2004 A edition) to confirm common peaks. Cluster analysis and principal component analysis were performed by using SPSS 19.0 software.RESULTS:The similarities were more than 0.95 in HPLC chromatograms of 15 batches of C. annuum. There were 12 common peaks. Its HPLC fingerprint was in good agreement with that of control fingerprint. 15 batches can be divided into three sub-categories as S1,S3-S5,S7,S9-S13 sub-categorie,S2,S14,S15 sub-categorie and S6,S8 sub-categorie. Through the principal component analysis,the cumulative contribution rate of 4 main component factors was 94.093,and comprehensive score of S5 was the highest with the best quality. CONCLUSIONS:Established HPLC fingerprints,cluster analysis and principal component analysis results can provide reference for the quality control of C. annuum.
OBJECTIVE:To establish HPLC fingerprint of Capsicum annuum,and to conduct cluster analysis and principal component analysis. METHODS:HPLC method was adopted. The determination was performed on Agilent C_(18) column with mobile phase consisted of methanol-0.2% phosphoric acid solution(gradient elution) at the flow rate of 1.0 mL/min. The detection wavelength was set at 235 nm,and column temperature was 30 ℃. The sample size was 15 μL. Using capsaicin peak as reference,HPLC fingerprints of 15 batches of C. annuum from different production areas were determined. The similarity evaluation ofcommon peaks was evaluated by using the TCM Chromatographic Fingerprint Similarity Evaluation System(2004 A edition) to confirm common peaks. Cluster analysis and principal component analysis were performed by using SPSS 19.0 software.RESULTS:The similarities were more than 0.95 in HPLC chromatograms of 15 batches of C. annuum. There were 12 common peaks. Its HPLC fingerprint was in good agreement with that of control fingerprint. 15 batches can be divided into three sub-categories as S1,S3-S5,S7,S9-S13 sub-categorie,S2,S14,S15 sub-categorie and S6,S8 sub-categorie. Through the principal component analysis,the cumulative contribution rate of 4 main component factors was 94.093,and comprehensive score of S5 was the highest with the best quality. CONCLUSIONS:Established HPLC fingerprints,cluster analysis and principal component analysis results can provide reference for the quality control of C. annuum.
引文
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[11]冯华,刘英波,刘亮,等.黔产辣蓼及其混淆品高效液相指纹图谱研究[J].中草药,2015,46(19):2943-2945.
[12]张晶,孙长波,石磊岭,等.RP-HPLC法测定辣椒中辣椒素、二氢辣椒素和降二氢辣椒素含量[J].药物分析杂志,2011,31(2):244-246.
[13]付文婷,詹永发,何建文,等.10个贵州地方辣椒品种品质评价[J].中国瓜菜,2018,31(12):37-40.
[14]张建,杨瑞东,陈蓉,等.贵州遵义辣椒矿质元素含量与其品质相关性分析[J].食品科学,2018,39(10):215-220.
[15]封兴智,詹永发,杨红,等.遵义市辣椒产业发展的现状、问题与对策[J].辣椒杂志,2007,5(2):5-7、10.
[2]贵州省食品药品监督管理局.贵州省中药材、民族药材质量标准[S].2003年版.贵阳:贵州科技出版社,2003:406.
[3]梁晨,韩盛玺.辣椒素临床应用的研究进展[J].华西药学杂志,2008,23(2):185-187.
[4]蒋海清,侯奕,黄晓焰,等.辣椒治疗胃黏膜损伤及溃疡的实验与临床研究[J].赣南医学院学报,2003,23(4):369-372.
[5]张晶,佟全胜,石磊岭,等.辣椒的化学成分研究进展[J].中成药,2009,31(12):1906-1912.
[6]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.
[7]冯华,石尚友,罗秀琼,等.黔产大菟丝子药材高效液相色谱指纹图谱的鉴别研究[J].时珍国医国药,2017,28(3):634-635.
[8]冯华,石尚友,罗秀琼,等.黔产见血清药材薄层色谱与HPLC指纹图谱识别[J].时珍国医国药,2017,28(10):2423-2425.
[9]朱星宇,陈勇强.SPSS多元统计分析方法及应用[M].北京:清华大学出版社,2011:241-247.
[10]范莉,侯小龙,王文清,等.指纹图谱结合一测多评模式在藤茶质量评价中的应用研究[J].中草药,2016,47(22):4076-4081.
[11]冯华,刘英波,刘亮,等.黔产辣蓼及其混淆品高效液相指纹图谱研究[J].中草药,2015,46(19):2943-2945.
[12]张晶,孙长波,石磊岭,等.RP-HPLC法测定辣椒中辣椒素、二氢辣椒素和降二氢辣椒素含量[J].药物分析杂志,2011,31(2):244-246.
[13]付文婷,詹永发,何建文,等.10个贵州地方辣椒品种品质评价[J].中国瓜菜,2018,31(12):37-40.
[14]张建,杨瑞东,陈蓉,等.贵州遵义辣椒矿质元素含量与其品质相关性分析[J].食品科学,2018,39(10):215-220.
[15]封兴智,詹永发,杨红,等.遵义市辣椒产业发展的现状、问题与对策[J].辣椒杂志,2007,5(2):5-7、10.