基于UPLC指纹图谱结合色度值的槐花炮制前后变化规律研究

Analysis of Changes of Sophorae Flos before and after Processing Based on UPLC Fingerprint and Chromatic Value

    摘要
  • 摘要:
      目的  分析槐花炮制前后UPLC指纹图谱与色度值的差异及动态变化规律, 为槐花生品、炒槐花、槐花炭的鉴别及炮制工艺研究提供参考。
      方法  建立槐花炮制前后样品的UPLC指纹图谱, 采用分光测色仪测定色度值, 结合相似度评价、单因素方差分析、聚类分析、主成分分析以及正交偏最小二乘法判别分析(OPLS-DA)等方法, 研究槐花生品、炒槐花、槐花炭的差异性, 同时分析槐花炮制过程中指纹图谱与色度值的动态变化规律。
      结果  槐花炮制前后指纹图谱和色度值差异明显。槐花生品、炒槐花、槐花炭分别标定了15、16、14个共有峰, 指认了峰2、峰3、峰8、峰9、峰10、峰11、峰14、峰15分别为5-羟甲基糠醛、原儿茶酸、芦丁、异槲皮苷、山柰酚-3-O-芸香糖苷、水仙苷、槲皮素、山柰素, 其中峰1和峰2(5-羟甲基糠醛)为炮制后新增的特征峰, 峰17则在炮制后缺失; 槐花炮制前后色度值发生明显变化, 其中槐花炭的色度值L*、b*、E*远低于槐花生品和炒槐花, 炒槐花的色度值a*稍高于槐花生品和槐花炭, 依据色度值可区分槐花生品、炒槐花和槐花炭样品; 聚类分析和主成分分析均可将槐花生品、炒槐花和槐花炭进行区分; OPLS-DA结果表明, 芦丁和槲皮素是槐花生品、炒槐花、槐花炭主要的差异成分(VIP值>1)。槐花生品在炮制46 min内, 峰1~峰16的峰面积/称样量/1 000比值均呈现先增大后减小的规律, 峰17则呈现一直减小的规律。以饮片色泽为评价指标, 炒制温度为160℃时, 炒槐花、槐花炭的炒制时间应分别以4~6、14~18 min为宜。
      结论  建立的UPLC指纹图谱和色度值的测定方法可为槐花生品、炒槐花、槐花炭的鉴别以及炮制工艺研究提供参考。

     

    Abstract:
      OBJECTIVE  To analyze the difference and dynamic change rules of UPLC fingerprint and chromatic value of Sophorae Flos before and after processing, and to provide reference for the identification and processing technology of Sophorae Flos, fried Sophorae Flos, and Sophorae Flos carbonisatus.
      METHODS  The UPLC fingerprint of Sophorae Flos before and after processing was established, and chromatic value was determined by spectrophotometer.The differences of Sophorae Flos, fried Sophorae Flos, and Sophorae Flos carbonisatus were analyzed according to the similarity evaluation, one-way ANOVA, cluster analysis, principal component analysis, orthogonal partial least squares discriminant analysis (OPLS-DA), et al.The dynamic change rule of the fingerprint and chromatic value of Sophorae Flos at different processing time points were also analyzed.
      RESULTS  There were obvious differences in fingerprints and chromatic value before and after processing.15 common peaks were calibrated for Sophorae Flos, 16 for fried Sophorae Flos, and 14 for Sophorae Flos carbonisatus.Peak 2, 3, 8, 9, 10, 11, 14, and 15 were identified as 5-HMF, protocatechuic acid, rutin, isoquercetin, kaempferol-3-O-rutinoside, narcissin, quercetin, and kaempferol, respectively.Peak 1 and peak 2(5-HMF) were the chemical components produced after processing and peak 17 was the component disappeared after processing.There was a significant change in the chromaticity value before and after the processing of Sophorae Flos.The chromaticity values L*, b*, E* of Sophorae Flos carbonisatus were much lower than those of Sophorae Flos and fried Sophorae Flos.The chromaticity value a* of fried Sophorae Flos was slightly higher than that of Sophorae Flos and Sophorae Flos carbonisatus.It was possible to distinguish samples of Sophorae Flos, fried Sophorae Flos, and Sophorae Flos carbonisatus by the chromaticity value.Cluster analysis and principal component analysis could distinguish Sophorae Flos, fried Sophorae Flos, and Sophorae Flos carbonisatus.The results of orthogonal partial least squares discriminant analysis show that rutin and quercetin were the important components causing the difference between Sophorae Flos and its processed products (VIP>1).Within 46 min of processing of Sophorae Flos, the ratio of peak area/scale sample size/1 000 of peak 1 to peak 16 increased first and then decreased, while peak 17 decreased continuously.Using the color of the slices as an evaluation indicator when the frying temperature was 160℃, the appropriate frying time for fried Sophorae Flos, Sophorae Flos carbonisatus should be 4-6 min and 14-18 min, respectively.
      CONCLUSION  The established UPLC fingerprint and chromatic value determination method can provide reference for the identification and processing technology of Sophorae Flos, fried Sophorae Flos, and Sophorae Flos carbonisatus.

     

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