HPLC同时测定紫斑牡丹花粉中槲皮素、木犀草素、异鼠李素的含量

Simultaneous Determination of Quercetin,Luteolin and Isorhamnetin in Pollen of Paeonia Rockii by RP-HPLC

关键词:  紫斑牡丹花粉  槲皮素  木犀草素  异鼠李素  含量测定  高效液相色谱法

Key words:  pollen of paeonia rockii  quercetin  luteolin  isorhamnetin  content assay  HPLC

摘要:

目的 建立同时测定紫斑牡丹花粉中3个黄酮类化合物槲皮素、木犀草素、异鼠李素含量的RP-HPLC。方法 样品用25倍75%乙醇回流提取2 h，提取液加15%盐酸水解2.5 h；采用Agilent Eclipse plus C₁₈色谱柱（250 mm×4.6 mm，5 μm），以甲醇-乙腈-0.1%磷酸（25：13：62）为流动相等度洗脱，流速：1.0 mL·min^-1，检测波长：360 nm，柱温35℃。结果 槲皮素、木犀草素和异鼠李素分别在1.3~26.0，0.7~14.0，8.4~168.0μg·mL^-1内呈良好的线性关系，相关系数均≥0.9999；加样回收率（n=9）的RSD值分别为99.8%，100.3%，100.8%。结论 该方法简单、高效，可作为紫斑牡丹花粉质量控制的参考

Abstract:

OBJECTIVE To develope a RP-HPLC method for simultaneous determination of three flavonoids (quercetin, luteolin, isorhamnetin) in pollen from Paeonia rockii.METHODS The samples were pretreated, including reflux extraction with 75% ethanol for 2 h(with the ratio of liquor to material of 25:1), and then the extracting solution was hydrolyzed with 15% hydrochloric acid for another 2.5 h. Agilent Eclipse plus C₁₈ column (250 mm×4.6 mm, 5 μm) was adopted. The mobile phase was methanol-acetonitrile-0.1% phosphoric acid (25:13:62) at the flow rate of 1.0 mL·min^-1. The detection wavelength was 360 nm and the column temperature was 35℃.RESULTS Calibration curves were found to be linear in the range of 1.3-26.0 μg·mL^-1 for luteolin, 0.7-14.0 μg·mL^-1 for quercetin and 8.4-168.0 μg·mL^-1 for isorhamnetin. The correlation coefficients were greater or equal to 0.999 9. The average recoveries(n=9) of three flavonoids were 99.8%, 100.3% and 100.8%, respectively.CONCLUSION The developeded method is simple and efficient, which can be used for quality control of pollen of Paeonia rockii.