HPLC指纹图谱和多成分定量结合化学模式识别法评价不同生长期刺山柑果质量

HPLC fingerprint and multi-components determination combined withchemical pattern recognitionto evaluate quality offruits of Capparis spinosaat different growth stages

目的建立不同生长期刺山柑Capparis spinosa果实的HPLC指纹图谱及多成分含量测定,结合化学模式识别法评价不同生长期刺山柑果的质量,为其进一步研究和开发提供依据。方法采用Agilent ZORBAX SB-C18色谱柱,以0.1%磷酸溶液(A)-乙腈(B)为流动相进行梯度洗脱,体积流量为1.0 mL/min,检测波长为310 nm,柱温为25℃,进样量为10μL。采用《中药色谱指纹图谱相似度评价系统(2012版)》建立18批次不同生长期刺山柑果的HPLC指纹图谱并分析相似度。使用SPSS 25.0和SIMCA-P 14.1软件进行聚类分析、主成分分析和偏最小二乘法-判别分析(partial least squares-discriminant analysis,PLS-DA)。通过对照品比对指认化学成分并进行定量测定。结果18批次刺山柑果HPLC指纹图谱共匹配出14个共有峰,分别指认出2号峰为原儿茶酸、13号峰为芦丁、14号峰为山柰酚-3-O-芸香糖苷,指纹图谱相似度范围为0.924~0.991。聚类分析将18批次刺山柑果分为3类;主成分分析得刺山柑成熟果质量较好;经PLS-DA分析筛选出了芦丁和山柰酚-3-O-芸香糖苷等4个差异成分。样品中原儿茶酸、芦丁、山柰酚-3-O-芸香糖苷含量分别为0.0323~0.0594、0.1157~0.4640、0.0629~0.3307 mg/g。结论建立的刺山柑果HPLC指纹图谱操作简便、结果可靠,原儿茶酸、芦丁和山柰酚-3-O-芸香糖苷可以作为刺山柑果质量评价的指标性成分。

ObjectiveTo establish HPLC fingerprints and multi-component content determination for fruits of Capparis spinosain different growth stages,and to evaluate the quality for fruits of C.spinosain different growth stages in combination with chemical pattern recognition,so as to provide a basis for further research and development.Methods Agilent ZORBAX SB-C18column was used for gradient elution with 0.1%phosphoric acid solution(A)-acetonitrile(B)as the mobile phase at a flow rate of 1.0 mL/min,a detection wavelength of 310 nm,a column temperature of 25oC,and an injection volume of 10μL.The Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 Edition)was used to establish the HPLC fingerprints of 18 batches for fruits of C.spinosawith different growth stages and analyze the similarity.Clustering,principal component analysis,and partial least squares-discriminant analysis(PLS-DA)were performed using SPSS 25.0 and SIMCA-P 14.1 software.Chemical composition was identified by comparison of controls and the content in the sample was determined.Results A total of 14 common peaks were matched by the HPLC fingerprint of 18 batches for fruits of C.spinosa,and peak 2 was identified as protocatechuic acid,peak 13 as rutin,and peak 14 as kaempferol-3-O-rutinoside,and the fingerprint similarity range was 0.924 to 0.991.Cluster analysis divided 18 batches for fruits of C.spinosainto three categories;Principal component analysis showed that the quality of ripe for fruits of C.spinosawas better;Four differentiated components,including rutin and kaempferol-3-O-rutinoside,were screened by PLS-DA analysis.The contents of protocatechuic acid,rutin and kaempferol-3-O-rutinoside were 0.0323-0.0594,0.1157-0.4640,0.0629-0.3307 mg/g.Conclusion The established HPLC fingerprint for fruits of C.spinosais easy to operate and the results are reliable,and protocatechuic acid,rutin and kaempferol-3-O-rutinoside can be used as an indicator component for the quality evaluation for fruits of C.spinosa.