莱菔子炮制前后HPLC特征图谱及4种成分含量变化研究

Study on Characteristic Fingerprint of Raw and Processed Raphani Semen and Changes of Four Chemical Components

目的建立莱菔子生品和炒制品的特征图谱并测定其特征成分含量,探讨莱菔子炮制前后化学成分变化规律,为合理控制莱菔子质量提供参考。方法采用HPLC对12批莱菔子炮制前后样品进行研究,采用《中药色谱指纹图谱相似度评价系统》(2012版)建立莱菔子生品和炒制品特征图谱并进行相似度评价,对其特征图谱峰面积进行聚类分析(HCA)和正交偏最小二乘判别分析(OPLS-DA)。结果莱菔子生品和炒制品特征图谱分别标定出8、11个共有峰,12批莱菔子生品和炒制品与对照图谱相似度均大于0.95。HCA和OPLS-DA可明显区分生品和炒制品,萝卜硫苷特征峰标识性最强。生品中未检出萝卜硫苷,芥子碱硫氰酸盐、莱菔素和芥子酸含量分别为0.21%~0.36%、0.80%~1.61%、0.02%~0.05%;炒制品中萝卜硫苷、芥子碱硫氰酸盐含量分别为2.70%~4.52%、0.29%~0.52%,莱菔素和芥子酸均未检出。结论莱菔子生品和炒制品特征图谱专属性强、差异显著,结合特征成分含量测定可有效区分莱菔子生品和炒制品并控制其质量。

Objective To establish HPLC characteristic fingerprint and determine the characteristic chemical components in raw and processed Raphani Semen;To explore the changes of the chemical components in raw and processed Raphani Semen;To provide reference for the rational quality control of Raphani Semen.Methods HPLC analysis was performed to evaluate 12 batches of samples of raw and processed Raphani Semen.Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012 edition)was used for establishing the reference fingerprint and similarity evaluation.Hierarchical clustering analysis(HCA)and orthogonal partial least squares-discriminate analysis(OPLS-DA)were used in analysis of characteristic spectrum peak area.Results The HPLC fingerprint of raw and processed Raphani Semen was constructed with 8 and 11 common chromatographic peaks,respectively.The similarities of 12 batches of raw and processed Raphani Semen were all over 0.95 compared with the reference fingerprint.The raw and processed products could be clearly distinguished by HCA and OPLS-DA,and the characteristic peak of glucoraphanin was the most discriminative.Glucoraphanin was not detected in raw products.The contents of sinapine thiocyanate,sulforaphene and sinapic acid in raw products were 0.21%–0.36%,0.80%–1.61%,0.02%–0.05%,respectively.The contents of glucoraphanin and sinapine thiocyanate in processed products were 2.70%–4.52%and 0.29%–0.52%,respectively.Neither sulforaphane nor sinapic acid was detected in processed products.Conclusion The HPLC fingerprint of raw and processed Raphani Semen have strong specificity and significant differences.The characteristic fingerprint combined with determination of characteristic chemical components could be used for effectively distinguish and control the quality of raw and processed Raphani Semen.