紫斑牡丹花粉HPLC特征图谱及4个成分含量测定

HPLC characteristic chromatogram and assay of four components for pollen of Paeonia rockii

目的:利用高效液相色谱法建立紫斑牡丹花粉的特征图谱,并测定不同产地紫斑牡丹花粉中氧化芍药苷、芍药内酯苷、芍药苷、槲皮素4个成分的含量。方法:采用CAPCELL PAK-C18(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.2%甲酸水(B)为流动相梯度洗脱,流速1 mL·min^-1,检测波长274nm,柱温30℃;采用中药色谱指纹图谱相似度评价软件(2012年版)对13批牡丹花粉进行相似度分析,同时测定7批不同产地牡丹花粉中4个成分的含量。结果:13批紫斑牡丹花粉样品得到的指纹图谱相似度大于0.960,标定共有峰16个,并确定了其中4个色谱峰的归属。产自甘肃兰州、安徽亳州、山东菏泽的7批紫斑牡丹花粉样品中氧化芍药苷、芍药内酯苷、芍药苷、槲皮素的含量分别在2.488~7.675、0~15.251、2.339~10.433、6.462~15.924 mg·g^-1之间。结论:HPLC指纹图谱结合定量测定能全面反映紫斑牡丹花粉内在质量,该方法简便可行,快速准确,可为其质量评价提供参考。

Objective:To establish the HPLC characteristic chromatogram of Pollen of Paeonia rockii and to develop the simultaneous determination method of oxypaeoniflora,albiflorin,paeoniflorin and quercetin in the samples.Methods:HPLC analysis was performed on a CAPCELL PAK-C18 column(250 mm×4.6 mm,5μm).The mobile phase consisted acetonitrile(A)and 0.2%formic acid aqueous solution(B)with gradient elution.The flow rate was 1 mL·min^-1,the detection wavelength was 274 nm and the column temperature was 30℃.TCM chromatographic fingerprint similarity evaluation software(2012 version)was used for similarity analysis of 13 batches of Paeonia rockii pollen,and the contents of four components in 7 batches of Paeonia rockii pollen from different habitats were determined at the same time.Results:The similarities of the chromatographic fingerprint of 13 batches of Paeonia rockii pollen samples were above 0.960.There were altogether 16 common peaks,and four of them were identified.The contents of oxypaeoniflora,albiflorin,paeoniflorin and quercetin in 7 batches of samples from Lanzhou,Bozhou and Heze city were in the range of 2.488-7.675,0^-15.251,2.339^-10.433 and 6.462^-15.924 mg·g^-1,respectively.Conclusion:The combination of HPLC fingerprint and content determination can reflect inherent quality of Paeonia rockii pollen.The proposed method is simple,rapid and accurate,providing reference for quality evaluation of the drug.