基于化学模式识别技术的枳实HPLC定量指纹图谱研究

建立了枳实的高效液相色谱(HPLC)指纹图谱分析方法。色谱柱为Tnature-ACCHROM C18色谱柱(4.6 mm×250 mm,5μm);以乙腈-0.5%甲酸水溶液为流动相进行梯度洗脱,结合液相色谱-四极杆飞行时间质谱(HPLC-QTOF-MS)联用技术对枳实指纹图谱中的共有峰进行鉴定;采用相似度评价、聚类分析(CA)、主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)对22批枳实进行数据分析及质量评价。结果显示:指纹图谱共标定12个共有峰,HPLC-QTOF-MS分析指认出11个成分;22批枳实样品的相似度在0.9以上;CA、PCA和OPLS-DA的分析结果一致,其中江西产地聚为一类,湖南和福建产地聚为一类,并筛选出橙皮苷、新橙皮苷和柚皮苷3个差异性质量标志物。所建立的枳实HPLC指纹图谱方法稳定、可靠,可为其质量控制提供参考依据。

An HPLC Fingerprint Research on Citrus aurantium Based on Chemical Pattern Recognition Technology

A fingerprint analysis method was developed for the quality control of Citrus aurantium by high performance liquid chromatography(HPLC).The separation was carried out on a Waters Tnature-ACCHROM C18(4.6 mm×250 mm,5μm)column,using acetonitrile-0.5%formic acid water as mobile phase by gradient elution,and the common peaks in the fingerprint of Citrus aurantium were identified by high performance liquid chromatography-quadrupole time of flight-mass spectrometry(HPLC-QTOF-MS).Similaritity evaluation,cluster analysis(CA),principal component analysis(PCA)and orthogonal partial least square discriminant analysis(OPLS-DA)were used for the data analysis and quality evaluation on 22 batches of Citrus aurantium.Results showed that 12 common peaks were defined by the fingerprint,and 11 components were identified by HPLC-QTOF-MS.The similarities for 22 batches of Citrus aurantium samples were more than 0.9.The results obtained from CA,PCA and OPLS-DA were consistent,among which,the samples from Jiangxi Province were clustered into one group,while the samples from Hunan Province and Fujian Province were clustered into another group,and three quality differential markers were screened out as hesperidin,neohesperidin and naringin by OPLS-DA.The HPLC fingerprint for Citrus aurantium is stable and reliable,and could be used to provide a reference for its quality control.