基于指纹图谱、模式识别及多成分定量的生稻芽质量评价

    Quality Evaluation of raw Oryzae Fructus Germinatus Based on Fingerprint, Pattern Recognition and Multi-components Determination

    • 摘要:
      目的 建立生稻芽HPLC指纹图谱,结合化学模式识别技术对其进行分析,并测定生稻芽中主要次生代谢成分5-羟甲基糠醛、原儿茶酸、香草酸、对香豆酸、阿魏酸的含量,为质量评价提供一定的科学依据。
      方法 采用 Ultimate AQ - C18 (250 mm×4.6 mm,5 μm)色谱柱,以乙腈-0.3%磷酸溶液进行梯度洗脱,建立16批生稻芽的HPLC指纹图谱,通过中药色谱指纹图谱相似度评价系统,结合聚类分析和主成分分析对指纹图谱进行数据分析,同时测定 5种有效成分含量。
      结果 建立了16批次生稻芽HPLC指纹图谱, 相似度均≥0.891,确定18个共有峰,指认出5个色谱峰,分别为3号峰5-羟甲基糠醛、4号峰原儿茶酸、10号峰香草酸、17号峰对香豆酸及18号峰阿魏酸;聚类分析和主成分分析结果一致,将16批样品分为4类。多成分定量分析方法学考察结果表明,5个指标成分在各自范围内的线性关系良好(R2≥0. 9993),加样回收率为 99.12%~101.37%,RSD为0.02%~2.40%;16批生稻芽中5-羟甲基糠醛、原儿茶酸、香草酸、对香豆酸、阿魏酸的含量分别为0.36~3.57,1.42~23.36,6.60~39.37,33.02~72.33,29.72~72.59 μg·g−1
      结论 HPLC指纹图谱结合模式识别及多成分同时测定的方法快速、简便、重复性好,可为生稻芽的质量评价提供一定的理论参考。

       

      Abstract:
      OBJECTIVE To establish the fingerprint of raw Oryzae Fructus Germinatus by HPLC, analyze them with chemical pattern recognition technology, and determine the contents of 5-hydroxymethylfurfural, protocatechuic acid, vanillic acid, p-coumaric acid and ferulic acid in raw Oryzae Fructus Germinatus, in order to provide a scientific basis for its quality evaluation.
      METHODS The HPLC fingerprint of raw Oryzae Fructus Germinatus was performed on Ultimate AQ-C18(250 mm×4.6 mm, 5 μm) column with acetonitrile-0.3% phosphoric acid solution for gradient elution, the fingerprint data was analyzed by the similarity evaluation system of traditional Chinese medicine chromatographic fingerprints combined with cluster analysis and principal component analysis. Furthermore, the contents of five components in raw Oryzae Fructus Germinatus were determined.
      RESULTS The HPLC fingerprint of 16 batches of raw Oryzae Fructus Germinatus was established. The similarity were all≥0.891. Eighteen common peaks were marked and 5 of them were identified, including 5-hydroxymethylfurfural(peak 3), protocatechuic acid(peak 4), vanillic acid(peak 10), p-coumaric acid(peak 17) and ferulic acid(peak 18). Cluster analysis and principal component analysis were used to classify the 16 batches of raw Oryzae Fructus Germinatus into four groups. The results of multi-component quantitative analysis methodology showed that the linear relationship(R20.9993) of five components were good, the average recoveries were between 99.12% and 101.37%, and the RSD were in the range of 0.02%−2.40%. The contents of 5-hydroxymethylfurfural, protocatechuic acid, vanillic acid, p-coumaric acid and ferulic acid in 16 batches of raw Oryzae Fructus Germinatus were in the range of 0.36−3.57, 1.42−23.36, 6.60−39.37, 33.02−72.33, 29.72−72.59 μg·g−1, respectively.
      CONCLUSION The method of HPLC fingerprint combined with pattern recognition and multi-components simultaneous determination is rapid, simple and reproducible, which can provide a reference for the quality evaluation of raw Oryzae Fructus Germinatus.

       

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