HPLC-PDA指纹图谱结合UFLC-Q-TOF/MS定性鉴别评价不同产地白术药材质量  

Qualitative evaluation of Atractylodis Macrocephalae Rhizoma from different habitats by HPLC-PDA fingerprint combined with UFLC-Q-TOF/MS qualitative identification

作　　者：孙学[1] 文红梅[1] 崔小兵[1] 陆兔林[1] 李伟[1] 单晨啸[1] 

机构地区：[1]南京中医药大学药学院,江苏南京210023

出　　处：《中草药》2016年第19期3494-3501,共8页Chinese Traditional and Herbal Drugs

基　　金：国家中医药管理局中医药行业科研专项(201307008);江苏省中药学优势学科开放课题(YS2012ZYX309)

摘　　要：目的 建立白术药材HPLC-PDA指纹图谱并进行定性鉴别,为全面控制白术质量提供参考。方法 白术加70%甲醇超声60 min;色谱分析采用Inertsil？ODS-SP色谱柱（150 mm×4.6 mm,5μm）,柱温40℃,体积流量1.0 m L/min,采用Waters 2998紫外检测器,检测波长为235 nm,流动相为水（A）-乙腈（B）,洗脱梯度为0～10 min,30%～45%B;10～25 min,45%B;25～50 min,45%～70%B;50～55 min,70%B;55～62 min,70%～30%B;62～75 min,30%B。质谱测定采用电喷雾离子源（ESI）的飞行时间质谱（TOF/MS）;正离子模式下;质量扫描范围m/z 50～1 500。结果 分别对不同产地的白术进行比较、拟合,标定了白术HPLC-PDA指纹图谱的6个共有峰,并通过高分辨UFLC-Q-TOF/MS对共有峰进行了指认,分别为5-羟甲基糠醛、白术内酯III、白术内酯I、白术内酯II、白术内酯VI和双白术内酯;对白术的色谱条件及系统适用性、提取条件进行了优化、考察,其精密度、稳定性、重复性RSD值均小于2%;测得的10批次样品与拟合指纹图谱相似度均大于0.95。结论 所建立的HPLC指纹图谱可作为规范白术的均一性和稳定性的质量控制手段。Objective To establish and identify the HPLC-PDA fingerprint of Atractylodis Macrocephalae Rhizoma （AMR） and provide a reference for the comprehensive control of the quality of AMR. Methods AMR was extracted with 70% methanol by sonicating for 60 min. The analysis of AMR extract was performed on Inertsil ODS-SP column （150 mm ×4.6 mm, 5μm）, column temperature was maintained at 40℃, flow rate was 1.0 mL/min, and detector was Waters 2998 UV detector with detection wavelength 235 nm. Mobile phase was acetonitrile （B）-water （A） with the elution gradient 0 -10 min, 30%--45% B, 10--25 min, 45% B, 25--50 min, 45%--70% B, 50--55 min, 70% B, 55--62 min, 70%--30% B, 62--75 m/n, 30% B. Time-of-flight mass spectrometer （TOF/MS） and electro-spray ion （ESI） source were used for the qualitative analysis in a positive ion mode, and mass scan range was m/z 50--1 500. Results Comparing and fitting the peaks of AMR from different habitats （Zhejiang, Anhui, and Hunan Provinces）, the HPLC-PDA fingerprint was set up with six common peaks, and they were identified by UFLC-Q-TOF/MS as 5-（hydroxymethyl）-2-furaldehyde, atractylenolide III, atractylenolide I, atractylenolide II, atractylenolide VI, and biatractylenolide. System suitability, extraction, and chromatographic conditions of AMR were optimized. RSD of accuracy, stability and repeatability was all less than 2%. Measuring ten batches and fitting fingerprint similarity, the values were all greater than 0.95. Conclusion The HPLC fingerprint can be used as standard uniformity and stability of quality control methods for AMR slice.

关 键 词：白术 HPLC-PDA 指纹图谱 UFLC-Q-TOF/MS 5-羟甲基糠醛 白术内酯III 白术内酯I 白术内酯II 白术内酯VI 双白术内酯 

分 类 号：R286.6[医药卫生—中药学]