牡丹皮不同炮制品UPLC指纹图谱及多成分含量测定研究

摘要

Objective To establish the ultra-high performance liquid chromatography (UPLC) fingerprints of Moutan Cortex and its processed products,and determine the contents of gallic acid,5-hydroxymethyl furfurol,4-hydroxybenzoic acid,oxypaeoniflorin,paeoniflorin,benzoic acid,1,2,3,4,6-O-pentagalloyl-glucose and paeonol,so as to provide basis for the improvement of quality standards.Methods The UPLC separation was carried out on a Zorbax Eclipse Plus-C18 column (2.1 mm×100 mm,1.8 μm),and the mobile phase was consisted of water containing 0.1% formic acid and methanol containing 0.05% formic acid with gradient elution at the flow rate of 0.2 mL/min.The column temperature was maintained at 30 ℃.Results The UPLC fingerprints of Moutan Cortex,Moutan Cortex Tostus,Moutan Cortex Carbonisatus were developed,respectively,and paeonol was employed as the reference peak.Seventeen peaks were selected as common peaks in fingerprints of Moutan Cortex and Moutan Cortex Tostus,respectively,and eight common peaks were selected for Moutan Cortex Carbonisatus.The similarity of fingerprints were above 0.944,except that of batch T2 of Moutan Cortex Carbonisatus was 0.857.In addition to benzoic acid,the contents of rest components varied in different degree before and after the processing of Moutan Cortex.Conclusion The established UPLC fingerprint and multi-components determination method was rapid,convenient and reproducible.In conclusion,they could directly reflect the differences of compositions between the Moutan Cortex and its processed products.%目的 建立牡丹皮及其炮制品的超高效液相(UPLC)指纹图谱,并测定没食子酸、5-羟甲基糠醛(5-HMF)、对羟基苯甲酸、氧化芍药苷、芍药苷、苯甲酸、1,2,3,4,6-O-五没食子酰葡萄糖、丹皮酚等8个指标成分的质量分数,为牡丹皮药材质量标准的提高提供依据.方法 采用Zorbax Eclipse Plus-C18(2.1 mm×100 mm,1.8 μm)色谱柱,以质量分数0.1%甲酸水溶液-质量分数0.05%甲酸甲醇为流动相进行梯度洗脱,流速为0.2 mL/min,柱温为30 ℃,建立牡丹皮及其炮制品的UPLC指纹图谱,同时对以上8个成分进行测定.结果 建立了生丹皮、炒丹皮及丹皮炭3种药材的UPLC指纹图谱,以丹皮酚为参照峰,生丹皮和炒丹皮各标定了17个共有峰,丹皮炭标定了8个共有峰,除1批丹皮炭相似度为0.857外,样品与对照指纹图谱的相似度均大于0.944;除了苯甲酸的质量分数无明显变化外,其余化合物在牡丹皮炮制前后均有不同程度的增加或减少.结论 建立的UPLC指纹图谱及含量测定方法均具有较好的重现性,且分析时间短,方便快速,二者结合可直观地反映生丹皮、炒丹皮及丹皮炭3种药材之间成分的差异.