掌叶大黄UPLC多指标成分测定及指纹图谱研究

Study on multi-compound determination and fingerprint of Rheum palmatum by ultra performance liquid chromatography

目的:建立掌叶大黄超高效液相色谱(UPLC)多指标成分测定及指纹图谱方法。方法:采用C18色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-0.1%甲酸溶液梯度洗脱,流速0.3 mL·min^(-1),检测波长280 nm,测定10批掌叶大黄药材中儿茶素、1-O-没食子酰基-2-O-肉桂酰基-β-D-葡萄糖苷、大黄素、大黄酚和大黄素甲醚5个成分的含量,并建立掌叶大黄药材指纹图谱,采用超高效液相色谱-飞行时间质谱(UPLC-Q-TOF-MSE)技术对掌叶大黄的化学成分进行全面定性分析。结果:不同批次药材中5个成分含量差异较大;建立10批掌叶大黄药材UPLC指纹图谱,获得21个共有峰且相似度在0.936～0.985。通过对照品指认、文献比对以及高分辨质谱数据解析,初步鉴定了包括21个共有峰在内的71个化学成分。结论:建立的多成分测定结合指纹图谱分析方法,并利用液质联用技术的定性分析,为掌叶大黄药材的质量控制提供了可行的方法。

Objective：To establish a method for multi-compound determination and fingerprint of roots and rhizomes of Rheum palmatum by ultra performance liquid chromatography（UPLC）. Methods：UPLC separation was performed on a C18 column（100 mm×2.1 mm, 1.7 μm） by gradient elution with acetonitrile-0.1% formic acid. The flow rate was 0.3 mL·min^-1. The detection wavelength was 280 nm. The fingerprints and the contents of five compounds including（＋） -catechin, 2-O-cinnamoyl-1-O-galloyl-β-D-glucoside, emodin, chrysophanol and physcion in 10 batches of R. palmatum were analyzed. Ultra performance liquid chromatography tandem with time-of-flight mass spectrometer with MSE data-acquisition mode（UPLC-Q-TOF-MSE） was used to characterize the chemical constituents. Results：There was considerable difference in contents of five compounds in different batches of samples. The chromatographic fingerprints of 10 batches of the roots and rhizomes of R. palmatum were generated with 21 common peaks. The similarities between each sample and the reference fingerprint ranged from 0.936 to 0.985. A total of 71 components including 21 common peaks were putatively identified via referring to reference components, literature as well as analyzing their MS/MS data. Conclusion：The proposed UPLC quantitative analysis and fingerprint methods with qualitative analysis by UPLC-MS can be a reliable method for quality control of the roots and rhizomes of R. palmatum.