一种基于全波长UV建立HPLC指纹图谱的质量控制模式:以不同等级的玄参为例

Establishment of quality control model for high performance liquid chromatography fingerprint based on ultraviolet full-wavelength scanning spectrum:case study on different grades of Scrophulariae Radix

HPLC-UV指纹图谱是目前《中国药典》用于中药材质量控制的重要手段之一。然而,关于紫外(UV)具体的波段选择存在一定主观性,不能较好地表征中药内在的质量差异。因此,该研究以不同等级的玄参为研究对象,提出一种新的基于紫外全波长扫描图谱建立高效液相指纹图谱的质量控制模式。首先,采集不同等级的玄参样品,利用紫外光谱(200~400 nm)建立所有样品的紫外全波长吸收光谱。通过光谱数据的预处理和主成分分析(principal component analysis,PCA)以及偏最小二乘法判别分析(partial least squares discrimination analysis,PLS-DA)等多元统计分析方法建立差异分析模式,得到251 nm的紫外波段可能是区分不同等级玄参质量差异的主要贡献的波段。进一步,利用该波段建立其HPLC指纹图谱,通过多元统计方法分析可得,基于差异紫外波段筛选得到的HPLC指纹图谱较原来的UV指纹图谱有更显著的分类趋势,可用于判别不同等级的玄参样品,能够较好的体现各等级之间的差异性。该研究建立的方法对于中药特异性HPLC指纹图谱的建立及其质量控制具有较大的指导和参考意义。

High performance liquid chromatography-ultraviolet(HPLC-UV)fingerprint is one of the most important methods for the quality control of Chinese medicines in Chinese Pharmacopoeia.However,certain subjectivity is present in selection of specific band of UV,and the inherent quality differences of Chinese medicine can′t be well characterized by this method.Therefore,with different grades of Scrophulariae Radix were taken as the research object in this study,a new quality control model of HPLC-UV was established in this study based on the ultraviolet full-wavelength scanning spectrum.Firstly,different grades of Scrophulariae Radix samples were collected,and the full-wavelength ultraviolet absorption spectra of all the samples were established at the bands of 200-400 nm.In order to analyze the differences among samples,the analysis model was built following multivariate statistical analysis methods such as principal component analysis(PCA)and partial least squares discrimination analysis(PLS-DA)after the pretreatment of spectral data.The result showed that the ultraviolet band at 251 nm may contribute most to distinguish the quality differences among different grades of samples.Then,the HPLC fingerprints of samples were established with the band at 251 nm.The multivariate statistical analysis showed that there was a more significant classification trend in HPLC fingerprints than that in the original UV fingerprints,which could be used to distinguish different grades of samples,and could better reflect the differences among different grades.The method reported in this study can be of a great guidance and reference for the establishment of specific fingerprints of Chinese medicines as well as for the quality control of Chinese medicine.