HPLC指纹图谱技术结合灰色关联度法评价不同批次诃子质量

Evaluation of the Quality of Different Batches of Terminalia Chebula Retz. Based on HPLC Fingerprint Technology and Grey Relational Analysis

关键词:  诃子  指纹图谱  质量评价  灰色关联度

Key words:  Terminalia chebula Retz.  fingerprint  quality evaluation  grey relational analysis

摘要:

目的 测定10批次不同来源诃子药材的HPLC指纹图谱和4种指标成分，结合灰色关联度分析法建立其质量评价方法。方法 采用HPLC测定10批次不同来源诃子药材的指纹图谱及主要成分没食子酸、没食子酸甲酯、柯里拉京和诃子酸的含量。Agilent Eclipse Plus C₁₈色谱柱（4.6 mm×250 mm，5 μm），以0.03%磷酸水-甲醇为流动相，梯度洗脱，流速1 mL·min^-1，柱温30℃，测定波长270 nm，以"中药色谱指纹图谱相似度评价系统（2012版本）"建立指纹图谱。基于灰色关联度分析不同批次诃子各指标成分数据，以各评价单元序列相对于参考序列的相关关联度作为测度，对样品进行综合评价。结果 建立诃子样品的指纹图谱共标定19个共有峰，指认出其中3号峰为没食子酸，8号峰为没食子酸甲酯，10号峰为柯里拉京，19号峰为诃子酸；没食子酸、没食子甲酯、柯里拉京、诃子酸分别在5.625~180，2~68，25~800，5.718~183 μg线性关系良好（r分别为1，0.999 2，0.999 2，0.999 9，n=6），精密度、重复性和稳定性均良好；没食子酸、没食子甲酯、柯里拉京、诃子酸的平均加样回收率分别为97.92%，98.24%，100.35%，101.82%，RSD分别为2.33%，1.98%，2.19%，1.87%，大部分批次的诃子药材样品相似度达到要求。结论 灰色关联度分析法结合HPLC指纹图谱和多指标成分定量的评价方法简单、全面，可用于诃子的质量评价，为资源开发利用及优质种源的筛选提供了参考依据。

Abstract:

OBJECTIVE To establish the HPLC fingerprint and determine four index components of 10 batches of Terminalia chebula Retz. from different origins, and establish a quality evaluation method by grey relational analysis. METHODS The fingerprints of 10 batches of Terminalia chebula Retz. from different origins and the components of gallic acid, methyl gallate, corilagin and chebulinic acid were determined by HPLC. Agilent Eclipse Plus C₁₈ column(4.6 mm×250 mm, 5 μm) was adopted with 0.03% phosphoric acid water-methanol as mobile phase, gradient elution. The flow rate was 1 mL·min^-1, column temperature was 30℃, determination wavelength was 270 nm. The "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System(2012 version)" established a HPLC fingerprint. Based on the grey relational degree, the data of each index component of different batches were analyzed, and the relational degree of each evaluation unit sequence with respect to the reference sequence as a measure, and the sample was comprehensively evaluated. RESULTS The fingerprints of the samples were established to determine 19 common peaks. The peak number 3 was gallic acid, the peak number 8 was methyl gallate, the peak number 10 was corilagin, the peak number 19 was chebulinic acid, and the gallic acid was linear at 5.625-180 μg(r=1, n=6), methyl gallate had a good linear relationship between 2 μg and 68 μg(r=0.999 2, n=6), corilagin had a good linear relationship at 25-800 μg(r=0.999 2, n=6). The linear relationship of chebulinic acid at 5.718-183 μg(r=0.999 9, n=6), which precision, repeatability and stability were good. The average recovery rate of gallic acid methyl gallate, corilagin, chebulinic acid were 97.92%, 98.24%, 100.35%, 101.82%, RSD were 2.23%, 1.98%, 2.19%, 1.87%, respectively. The similarity of most batches of Terminalia chebula Retz. met the requirements. CONCLUSION The grey relational analysis combined with fingerprint and multi-index component quantitative evaluation method is simple and comprehensive, and can be used for the quality evaluation of Terminalia chebula Retz., which provides a reference for resource development and utilization and screening of high-quality Terminalia chebulia Retz..