基于指纹图谱结合多元统计分析的不同贮藏年限对广陈皮成分影响研究

Study on the Effect of Different Storage Life on the Composition of Citri Reticulatae Pericarpium Based on Fingerprint and Multivariate Statistical Analysis

目的 建立广陈皮HPLC指纹图谱及多成分含量测定方法,并结合多元统计分析,研究不同贮藏年限对广陈皮成分的影响。方法 色谱条件:采用Syncronis C_(18)色谱柱(4.6 mm×150 mm,5μm);流动相为乙腈(A)-0.05%磷酸(B),梯度洗脱;流速1.0 mL·min^(-1);柱温30℃;进样量5μL;检测波长330 nm。采用HPLC法建立10批次不同贮藏年限广陈皮药材的指纹图谱,标定其共有峰,进行相似度评价;运用聚类、主成分分析等多元统计方法,综合评价10批广陈皮药材成分变化差异;进一步对《中国药典》指标成分与差异色谱峰中的橙皮苷、甜橙黄酮、川陈皮素、橘红素与5-去甲川陈皮素等5种成分进行含量测定。结果 10批广陈皮药材指纹图谱相似度均大于0.990,共标定了19个色谱峰,说明不同贮藏年限广陈皮整体质量相对稳定;聚类分析将10批广陈皮分3类,主成分分析提取4个主成分,累积方差贡献率在89.760%,且通过对照品指认确定了其中4个差异色谱峰。含量测定橙皮苷、甜橙黄酮、川陈皮素、橘红素与5-去甲川陈皮素等成分含量范围分别为20.940-34.780 mg·g^(-1)、0.344-0.517 mg·g^(-1)、4.667-7.456 mg·g^(-1)、3.592-5.879 mg·g^(-1)、0.391-0.710 mg·g^(-1)。结论 建立的广陈皮指纹图谱和多成分含量测定方法简单、准确,具有良好的稳定性和重复性,可采用聚类分析与主成分分析等多元统计分析方法对不同贮藏年限广陈皮药材的成分变化及质量进行评价,为其质量控制和临床应用提供了参考。

Objective To establish the HPLC fingerprint and determination method of main components in Citri Reticulatae Pericarpium(CRP), combined with multivariate statistical analysis, to explore the effect of different storage life on the components of CRP. Methods The chromato-graphic conditions were as follows: Syncronis C_(18)column(4.6 mm×150 mm, 5 μm), mobile phase acetonitrile(A)-0.05% phosphoric acid(B), gradient elution, flow rate 1.0 mL·min^(-1), column temperature 30℃, injection volume 5 μL, detection wavelength 330 nm. The fingerprints of 10 batches of CPR with different storage life were established by HPLC, the common peaks were calibrated, and the similarity was evaluated by multivariate statistical analysis methods such as cluster analysis and principal component analysis, and the chromatographic peaks causing the differences were identified. Furthermore, the contents of hesperidin and differential chromatographic peak nobiletin, tangeretin, sinensetin and 5-demethylnobiletin were determined. Results The similarity of fingerprints of 10 batches of CRP was greater than 0.990. A total of 19 chromatographic peaks were calibrated, indicating that the overall quality of CRP was relatively stable in different storage years;Cluster analysis divided 10 batches of CRP into 3 categories, and four principal components were extracted by principal component analysis, with a cumulative variance contribution rate of 89.760%, and four different chromatographic peaks were identified by reference. The contents of hesperidin, nobiletin, tangeretin, sinensetin and 5-demethylnobiletin were 20.940-34.780 mg·g^(-1), 0.344-0.517 mg·g^(-1), 4.667-7.456 mg·g^(-1), 3.592-5.879 mg·g^(-1), 0.391-0.710 mg·g^(-1), respectively.Conclusion The established fingerprints and multi-component content determination methods of CRP are simple,accurate, and have good stability and repeatability. Multiple statistical analysis methods such as cluster analysis and principal component analysis can be used to evaluate the component changes and quality of CRP in different storage years, providing a reference for its quality control and clinical application.