论文部分内容阅读
摘 要 目的：建立金橘药材的超高效液相色谱（UPLC）指纹图谱，并进行聚类分析和主成分分析。方法：采用UPLC法，色谱柱为Waters Acquity UPLC BEH C18，流动相为乙腈-0.1%磷酸水溶液（梯度洗脱），流速为0.3 mL/min，检测波长为330 nm，进样量为2 μL。以金柑苷峰为参照，绘制8批药材样品的UPLC指纹图谱；采用《中药色谱指纹图谱相似度评价系统》（2012 版）进行相似度评价，确定共有峰；采用SPSS 24.0软件对8批药材样品进行聚类分析和主成分分析。结果：8批药材样品的UPLC指纹图谱有24个共有峰，相似度均大于0.97。聚类分析结果显示，8批药材样品可聚为两类，S1～S4、S6～S8聚为一类，S5聚为一类。经主成分分析，3个主成分因子的累计方差贡献率为81.366%。结论：所建UPLC指纹图谱及聚类分析和主成分分析结果可为金橘药材的质量控制提供参考。
关键词 金橘；超高效液相色谱法；指纹图谱；相似度评价；聚类分析；主成分分析
Establishment of UPLC Fingerprint， Cluster Analysis and Principal Component Analysis of Fortunella margarita
HUANG Huahua1，WANG Mingjun1，HUANG Mingqing2，LYU Shishi1，WANG Shengjiang1（1. Dept. of Pharmacy， Xiamen Medical College/Fujian Provincial Key Lab of Traditional Chinese Medicine/Fujian Provincial Key Lab of Traditional Chinese Medicine Finish Processing and Health Product Development， Fujian Xiamen 361023， China； 2. College of Pharmacy， Fujian University of TCM， Fuzhou 350122， China）
ABSTRACT OBJECTIVE： To establish UPLC fingerprint of Fortunella margarita， and to conduct its cluster analysis and principal component analysis. METHODS： UPLC method was adopted. The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution） at the flow rate of 0.3 mL/min. The detection wavelength was set at 330 nm， and sample size was 2 μL. Using fortunellin as reference， UPLC fingerprints of 8 batches of F. margarita were determined. The similarity of 8 batches of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System（2012 edition） to confirm common peak. Cluster analysis and principal component analysis were performed by using SPSS 24.0 software. RESULTS： There were 24 common peaks in UPLC fingerprints of 8 batches of sample，the similarity of which was higher than 0.97. Cluster analysis showed that 8 batches of samples were clustered into 2 categories. S1， S2， S3， S4， S6， S7 and S8 were clustered into one category； S5 was clustered into the other category. By principal component analysis， the accumulative contribution rate of three main components was 81.366%. CONCLUSIONS： Established UPLC fingerprint， the results of cluster analysis and principal component analysis can provide reference for quality control of F. margarita.
KEYWORDS Fortunella margarita； UPLC； Fingerprint； Similarity evaluation； Cluster analysis； Principal component analysis
金橘，别名金桔、山橘、金枣、金柑、金弹等，是芸香科植物金橘[Fortunella margarita（Lour.）Swingle]的果实，原产于我国，其中福建尤溪、广西融安、广西陽朔、江西遂川、湖南浏阳和浙江宁波是金橘的六大产区[1]。金橘药食同源，是一种营养价值较高的水果，同时也具有较高的药用价值[2]。《中华本草》记载，金橘性温，味辛、甘，具有理气解郁、消食化痰、醒酒之功效，主治胸闷郁结、脘腹痞胀、食滞纳呆、咳嗽痰多、伤酒口渴[3]。现代研究表明，金橘具有抑菌、抗氧化、调节免疫等作用[4-5]；除可直接食用或泡水饮用外，市面上还有咸金枣、金枣丹、金桔咀嚼片等相关制剂[6-7]。目前，金橘药用方面的研究主要集中在成分分离和药理活性方面[8-10]，未见质量控制方面的报道及相关质量标准。金橘药材中许多成分的对照品在市面上购买不到，目前仅有金柑苷对照品和野漆树苷对照品，但仅对一两个成分进行含量测定并不能有效控制金橘药材的整体质量。为此，本研究收集了我国主要产地的金橘药材样品，建立了其超高效液相色谱（UPLC）指纹图谱，并结合聚类分析和主成分分析进行综合质量评价，旨在为有效控制其质量提供参考。
关键词 金橘；超高效液相色谱法；指纹图谱；相似度评价；聚类分析；主成分分析
Establishment of UPLC Fingerprint， Cluster Analysis and Principal Component Analysis of Fortunella margarita
HUANG Huahua1，WANG Mingjun1，HUANG Mingqing2，LYU Shishi1，WANG Shengjiang1（1. Dept. of Pharmacy， Xiamen Medical College/Fujian Provincial Key Lab of Traditional Chinese Medicine/Fujian Provincial Key Lab of Traditional Chinese Medicine Finish Processing and Health Product Development， Fujian Xiamen 361023， China； 2. College of Pharmacy， Fujian University of TCM， Fuzhou 350122， China）
ABSTRACT OBJECTIVE： To establish UPLC fingerprint of Fortunella margarita， and to conduct its cluster analysis and principal component analysis. METHODS： UPLC method was adopted. The determination was performed on Waters Acquity UPLC BEH C18 column with mobile phase consisted of acetonitrile-0.1% phosphoric acid solution （gradient elution） at the flow rate of 0.3 mL/min. The detection wavelength was set at 330 nm， and sample size was 2 μL. Using fortunellin as reference， UPLC fingerprints of 8 batches of F. margarita were determined. The similarity of 8 batches of samples was evaluated by TCM Chromatographic Fingerprint Similarity Evaluation System（2012 edition） to confirm common peak. Cluster analysis and principal component analysis were performed by using SPSS 24.0 software. RESULTS： There were 24 common peaks in UPLC fingerprints of 8 batches of sample，the similarity of which was higher than 0.97. Cluster analysis showed that 8 batches of samples were clustered into 2 categories. S1， S2， S3， S4， S6， S7 and S8 were clustered into one category； S5 was clustered into the other category. By principal component analysis， the accumulative contribution rate of three main components was 81.366%. CONCLUSIONS： Established UPLC fingerprint， the results of cluster analysis and principal component analysis can provide reference for quality control of F. margarita.
KEYWORDS Fortunella margarita； UPLC； Fingerprint； Similarity evaluation； Cluster analysis； Principal component analysis
金橘，别名金桔、山橘、金枣、金柑、金弹等，是芸香科植物金橘[Fortunella margarita（Lour.）Swingle]的果实，原产于我国，其中福建尤溪、广西融安、广西陽朔、江西遂川、湖南浏阳和浙江宁波是金橘的六大产区[1]。金橘药食同源，是一种营养价值较高的水果，同时也具有较高的药用价值[2]。《中华本草》记载，金橘性温，味辛、甘，具有理气解郁、消食化痰、醒酒之功效，主治胸闷郁结、脘腹痞胀、食滞纳呆、咳嗽痰多、伤酒口渴[3]。现代研究表明，金橘具有抑菌、抗氧化、调节免疫等作用[4-5]；除可直接食用或泡水饮用外，市面上还有咸金枣、金枣丹、金桔咀嚼片等相关制剂[6-7]。目前，金橘药用方面的研究主要集中在成分分离和药理活性方面[8-10]，未见质量控制方面的报道及相关质量标准。金橘药材中许多成分的对照品在市面上购买不到，目前仅有金柑苷对照品和野漆树苷对照品，但仅对一两个成分进行含量测定并不能有效控制金橘药材的整体质量。为此，本研究收集了我国主要产地的金橘药材样品，建立了其超高效液相色谱（UPLC）指纹图谱，并结合聚类分析和主成分分析进行综合质量评价，旨在为有效控制其质量提供参考。