不同产地及引种栽培白芍药材指纹图谱研究

HPLC Fingerprint of Radix Paeoniae Alba from Different Habitats and Different Cultivated Species

目的建立白芍药材的高效液相色谱指纹图谱,为科学评价不同产地及引种栽培白芍药材的质量提供依据。方法采用HPLC-DAD方法,测定了44批白芍样品(含2批赤芍药材和5批引种栽培白芍药材)。色谱柱为Hypersil-C18柱(250 mm×4.6 mm i.d,5μm),流动相为乙腈和水(0.05%磷酸),梯度洗脱,流速1.0 ml/ml,柱温40℃,检测波长230nm,分析时间60 min,平衡时间14 min。结果确定了白芍药材中的14个共有峰,经相似度分析,可将白芍药材分成3类。不同质量等级白芍样品色谱指纹图谱中两个主要成分芍药苷和芍药内酯苷峰高比的变化规律如下:优质品在(5～10):1左右;一般品在(3.0～3.5):1之间;劣质品在(1.0～3.0):1之间。4批引种栽培白芍药材与其他主产地白芍药材指纹图谱的相似度均在0.90之上,且其共有峰的LC-MS测定结果与其它白芍药材一致,芍药苷和芍药内酯苷峰面积比为(5～10):1。结论引种栽培的白芍药材与主产地白芍药材质量一致,且更加稳定可控,因此可以作为白芍药材的新来源;建立的白芍指纹图谱方法稳定、可靠、重复性好,结合芍药苷和芍药内酯苷峰高比的变化规律,可应用于全面控制白芍药材的质量。

Objective To establish the HPLC chromatographic fingerprint analysis for the quality control of Radix Paeoniae Alba from different producing areas and different cultivated species. Methods HPLC-DAD method was applied to establish the chromatographic fingerprint of 44 batches of Radix Paeoniae Alba samples,（including 2 batches of Radix Paeoniae Rubra and 5 batches of cultivated Radix Paeoniae Alba）.Hypersil-C18（250 mm×4.6 mm i.d,5 μm）was used and gradient eluted with acetonitrile： water（acidified to 0.05% with phosphoric acid）,the flow rate was set at 1.0ml/ml and column temperature was set at 40℃,the detection wavelength was set at 230nm and the analysis time was 60min with an equilibrium time of 14min. Results The HPLC chromatographic fingerprint of Radix Paeoniae Alba showing 14 characteristic peaks was established.Based on the similarity analysis,all the samples could be divided into 3 categories.Furthermore,the peak height ratio of paeoniflorin to albiflorin was calculated and summarized as follows：（5 ～ 10）∶1 in the superior,（3.0 ～ 3.5）∶1 in the general,（1.0 ～ 3.0）∶1 in the inferior,respectively;four batches of cultivated Radix Paeoniae Alba showed good similarities with other samples from main producing areas and the coefficients were above 0.90.The LC-MS spectra of their common peaks were coincided with those of other Radix Paeoniae Alba sample. Conclusion The results indicated that the inherent quality of these cultivated species was similar with that of Radix Paeoniae Alba from main producing areas and more stable and controllable.Therefore,they can be used as new resources of Radix Paeoniae Alba.The established HPLC-DAD fingerprint method is repeatable,feasible and can provide the more intuitively evaluating on the quality of Radix Paeoniae Alba with the summarized variation pattern of the peak area ratio of paeoniflorin to albiflorin.