HPLC法测定清补平衡膏方中没食子酸、5-HMF和芍药苷的含量

Determination of Gallic Acid,5-Hydroxymethyl-2-furaldehyde and Paeoniflorin in Qingbu Pingheng Herbal Paste by HPLC

目的：建立同时测定清补平衡膏方中没食子酸、5-HMF和芍药苷含量的高效液相色谱法。方法：采用Hibar ODS柱（4.6 mm×250 mm,5μm）,以乙腈-0.1%甲酸溶液为流动相梯度洗脱,流速为1.0 m L·min-1,检测波长为230 nm,柱温35℃,进样量10μL。结果：没食子酸在13.45~672.75μg·m L-1（r=0.999 4）范围内,5-HMF在10.76~537.9μg·m L-1（r=0.999 8）范围内,芍药苷在17.69~884.61μg·m L-1（r=0.999 8）范围内,对照品进样浓度和色谱峰峰面积线性关系良好,加样回收率分别为99.80%、96.94%和99.46%,RSD值分别为2.91%、2.93%和2.97%。结论：建立的高效液相色谱法简单易行,结果准确可靠,可有效地用于清补平衡膏方的质量控制。

Objective： To establish the HPLC method for determining the contents of gallic acid,5-HMF and Paeoniflorin in Qingbu Pingheng herbal paste. Method： HPLC assay was carried out on a column of Hibar ODS（ 4. 6 mm × 250 mm,5 μm）. The mobile phase was acetonitrile-0.1% formic acid solution with gradient elution at a flow rate of 1 m L·min-1. The injection volume was 10 μL. The testing conditions were determined as detection wavelength in 230 nm and the column temperature was 35 ℃. Results： The linear range was 13.45 ~ 672.75 μg·m L-1（ r= 0.999 4） for gallic acid,10.76 ~ 537.9 μg·m L-1（ r= 0.999 8） for 5-HMF and 17.69~884.61 μg·m L-1（ r= 0.999 8） for paeoniflorin salvianolic acid B showed good linear correlations in the range of 7.812 ~ 250.0 μg·m L-1. The recycling rates were 99.80% for gallic acid with RSD = 2.91%,96.94% for 5-HMF with RSD = 2.93,99.46% for paeoniflorin with RSD = 2.97%. Conclusion：The method was simple,quick,reliable,accurate and very suitable to the quality control of Qingbu Pingheng herbal paste.