基于militarine含量结合UPLC指纹图谱的不同产地白及饮片质量分析

Quality analysis of Bletilae Rhizoma slices from different producing areas based on the militarine content determination and establishment of fingerprint by UPLC

关键词:  白及饮片  UPLC  含量测定  指纹图谱  质量分析

Key words:  Bletilae Rhizoma slice  UPLC  content determination  fingerprint  quality analysis

摘要:

[目的] 采用超高效液相色谱法（UPLC）测定militarine（1，4-二[4-（葡萄糖氧）苄基]-2-异丁基苹果酸酯）的含量，并建立不同产地白及饮片的UPLC指纹图谱，为白及饮片的质量控制提供参考依据。[方法] 采用ACQUITY UPLC^® BEH C₁₈色谱柱（2.1 mm×150 mm，1.7 μm），以乙腈-纯水为流动相进行梯度洗脱，流速0.3 mL/min，检测波长270 nm，柱温46℃，建立白及饮片的指纹图谱，进行相似度评价，在含量测定和指纹图谱基础上，采用聚类分析和主成分分析对不同产地白及饮片质量进行分析。[结果] 24批次的白及饮片militarine含量在18.98～44.87 mg/g之间，指纹图谱分析共标定12个共有峰，相似度在0.812～0.976之间，其中22批白及饮片的相似度均大于0.9，表明市售白及饮片整体质量基本一致；聚类分析和主成分分析将militarine含量和指纹图谱中4个关键成分的总相对含量均较低的样品聚为一类，聚类结果与相似度高低关系不大。[结论] 测定militarine含量结合UPLC指纹图谱评价白及饮片的质量，具有一定的科学性和可行性。

Abstract:

[Objcetive] Ultra High Performance Liquid Chromatography(UPLC) was used to determine the content of militarine and establish UPLC fingerprints of Bletilae Rhizoma slices from different producing areas,so as to provide reference for the quality control of Bletilae Rhizoma slices.[Methods] An ACQUITY UPLC^® BEH C₁₈ column (2.1 mm×150 mm,1.7 μm) was used to perform gradient elution with acetonitrile-pure water as the mobile phase. The flow rate was 0.3 mL/min,the detection wavelength was 270 nm,and the column temperature was 46℃. The fingerprints of Bletilae Rhizoma slices were established and similarity evaluation were performed. Based on the content determination and fingerprinting,cluster analysis and principal component analysis method were used to evaluate the quality of Bletilae Rhizoma slices from different producing areas.[Results] The content of militarine in 24 batches of Bletilae Rhizoma slices was ranging from 18.98~44.87 mg/g,and the fingerprint was established and 12 common peaks were identified. The similarity of these fingerprint was between 0.812~0.976. Notably,the similarity of 22 batches of Bletilae Rhizoma slices was greater than 0.9,which indicates that there is little difference in the overall quality of commercially available Bletilae Rhizoma slices. Moreover,samples with low content of militarine and 4 critical components in the fingerprint were grouped into one class by cluster analysis and principal component analysis,which had little relationship with the level of similarity.[Conclusion] The method of militarine content determination and establishment of fingerprint by UPLC used to evaluate the quality of Bletilae Rhizoma slice is scientific and feasible.