UPLC-PDA法测定蒲公英提取物中咖啡酸的含量

Determination of caffeic acid in dandelion extract by UPLC-PDA

关键词:  蒲公英提取物；超高效液相色谱-二极管阵列检测法；咖啡酸  含量测定

Key words:  dandelion extract  UPLC-PDA  caffeic acid  content determination

摘要:

建立超高效液相色谱-二极管阵列检测法（UPLC-PDA法）测定蒲公英提取物中咖啡酸的含量。采用反相高效液相色谱法,用带有二极管阵列检测器的高效液相色谱仪（UPLC-PAD）对咖啡酸进行色谱分离和快速筛查。以ACQUITY UPLCTMHSS T3色谱柱(2.1mm×100mm,1.8μm)为分离柱，柱温：35℃；流动相体系：A项为乙腈，B项为0.4%磷酸水溶液，进行梯度洗脱；流速：0.35mL/min；进样量：5μL；检测波长为323nm。通过光谱图、保留时间及峰面积参数对咖啡酸进行定性定量检测。咖啡酸在1～100 μg/mL的范围内线性良好（R2=0.999）；0.25 μg/mL咖啡酸对照品溶液与空白溶液的信噪比>3,为检出限，1 μg/mL 咖啡酸对照品溶液与空白溶液的信噪比>10,为定量限，完全满足检测需求。该方法色谱分离较好,分析速度较快,前处理简单,适用于蒲公英提取物中咖啡酸的定性定量检测。

Abstract:

This study was aimed to establish a high performance liquid chromatography-diode array(UPLC-PDA) test method for the determination of caffeic acid in dandelion extract. Caffeic acid can be separated and detected by reversed-phase high performance liquid chromatography (HPLC-PAD) with diode array detector. The test condition is like that: separate column is ACQUITY UPLCTMHSS T3 column (2.1mm × 100mm-1. 8 μ m),the column temperature is 35 ℃, mobile phase consists of acetonitrile and 0.4% phosphoric acid solution for gradient elution, the flow rate is 0. 35mL/ min, the injection volume is 5 μ L, and the detection wavelength is 323nm. Caffeic acid is detected qualitatively and quantitatively by the retention time and peak response in the chromatogram.The linearity of the method for caffeic acid detection complies to the requirement at the range of 1～100 μg/mL (R2=0.999).The limit of detection(LOD) is 0.25 μg/mL (S/N>3), and the limit of quantitation(LOQ) is 1μg/mL (S/N> 10)，which fully demonstrates the compliance of the test method. this method is suitable for qualitative and quantitative determination of caffeic acid in dandelion extract with fine separation, quick analysis and simple pretreatment procedure of the sample.