基于指纹图谱和多指标含量测定的蜜远志质量评价

Quality evaluation of honey-stir-baked Polygalae Radix based on combinative methods of fingerprint and quantitative analysis of multi-components

目的建立指纹图谱和多指标定量的蜜远志质量评价方法。方法Kromasil 100-5-C18色谱柱(250 mm×4.6 mm,5μm)为固定相,以乙腈(A)-0.2%磷酸水溶液(B)为流动相进行梯度洗脱,体积流量为1.0 mL/min,检测波长318 nm。对16批蜜远志建立指纹图谱,确定共有峰并进行相似度分析结合化学计量学分析。采用Waters Xbridge Shiled RP C18色谱柱(250 mm×4.6 mm,5μm)进行同时测定远志[口山]酮Ⅲ、3,6′-二芥子酰基蔗糖、细叶远志皂苷、远志酸、远志皂苷元的含量。结果在指纹图谱研究中,标定了27个共有峰,结合对照品和HPLC-Q-TOF-MS共指认8个成分,分别为西伯利亚远志糖A5、西伯利亚远志糖A6、远志[口山]酮IX、远志[口山]酮Ⅲ、3,6′-二芥子酰基蔗糖、细叶远志皂苷、远志酸、远志皂苷元。聚类分析将16批样品分为3类;经主成分分析,主成分1~3是影响蜜远志质量评价的主要因子。多指标含量测定中远志[口山]酮Ⅲ、3,6′-二芥子酰基蔗糖、细叶远志皂苷、远志酸、远志皂苷元的质量分数分别为0.027%~0.068%、0.074%~0.798%、1.1%~1.4%、0.15%~0.36%、0.15%~0.37%。经方法学验证,线性关系良好(r≥0.9996),平均加样回收率为97.15%~100.9%。结论所建立的指纹图谱结合多指标含量测定方法准确、高效,特征性强,可为蜜远志饮片的质量评价和临床应用提供有效参考。

Objective To establish HPLC fingerprints and multi-components determination method of Miyuanzhi(honey-stir-baked Polygalae Radix).Methods Kromasil 100-5-C18(250 mm×4.6 mm,5μm)column was used as stationary phase and acetonitrile-0.2%phosphoric acid aqueous solution was used as mobile phase for gradient elution,the flow rate was 1.0 mL/min and the detection wavelength was 318 nm.The fingerprints of 16 batches of Honey-stir-baked Polygalae Radix were established,the common peaks were determined,and the similarity analysis was carried out combined with stoichiometric analysis.The content of polygalaxanthone III,3,6′-disinapoyl sucrose,tenuifolin,polygalacic acid and senegenin were determined simultaneously by Waters Xbridge Shiled RP C18.Results In the fingerprint study,27 common peaks of Honey-stir-baked Polygalae Radix were demarcated and eight components were identified by reference and HPLC-Q-TOF-MS,which were sibiricose A5,sibiricose A6,polygalaxanthone IX,polygalaxanthone III,3,6′-disinapoyl sucrose,tenuifolin,polygalacic acid and senegenin,respectively.The 16 batches of Honey-stir-baked Polygalae Radix were divided into three categories by cluster analysis.According to the principal component analysis,the principal components 1—3 were the main factors affecting the quality evaluation of the samples.The mass fractions of polygalaxanthone III,3,6′-disinapoyl sucrose,tenuifolin,polygalacic acid and senegenin in multi-components determination were 0.027%—0.068%,0.074%—0.798%,1.1%—1.4%,0.15%—0.36%,0.15—0.37%,respectively.The method was verified,the linear relationship was good(r≥0.9996).The average recovery was 97.15%—100.9%.Conclusion The established HPLC fingerprints and multi-components determination method are accurate,efficient and characteristic,which can provide reference for the quality control and clinical application of Honey-stir-baked Polygalae Radix.