基于液相色谱的菊苣及其相关产品中6种功能成分的分析方法

Determination of Six Functional Components in Chicory and Its Related Products by Liquid Chromatography

菊苣酸、绿原酸、秦皮甲素、秦皮乙素、山莴苣素和山莴苣苦素是菊苣中6种具有特殊生物活性的物质,也是菊苣保健功能的重要指标之一。为建立高效液相色谱法(high-performance liquid chromatography,HPLC)和超高效液相色谱法(ultra high-performance liquid chromatography,UPLC)测定菊苣及其相关产品中6种功能成分含量的分析方法,采用70%的甲醇水溶液超声提取样品中的功能成分,以0.1%的磷酸水溶液和乙腈作为流动相,在HPLC法中使用SB-C18(4.6 mm×250 mm,5μm)色谱柱,UPLC法中采用填料粒径小的C18+色谱柱(2.1 mm×100 mm,1.5μm),流速分别为1.0 mL/min和0.4 mL/min,山莴苣素和山莴苣苦素的检测波长为258 nm,菊苣酸、绿原酸、秦皮甲素和秦皮乙素的检测波长为330 nm。该方法测定结果的相对标准偏差均小于4%,平均回收率在97.0%~103.6%。6种功能成分在HPLC法中的检出限为0.49 mg/kg~2.14 mg/kg,定量限为1.65 mg/kg~7.14 mg/kg,在UPLC法中检出限为1.11 mg/kg~5.24 mg/kg,定量限为3.72 mg/kg~17.50 mg/kg。

Chicoric acid,chlorogenic acid,esculin,aesculetin,lactucin and lactucopicrin are six compounds with special biological activities in chicory,which are important indices of the health care function of chicory.High-performance liquid chromatography(HPLC)and ultra high-performance liquid chromatography(UPLC)were employed for the determination of the six components in chicory and its related products.The functional components in the samples were extracted by ultrasonic extraction with 70%methanol aqueous solution.The gradient elution was carried out with 0.1%phosphoric acid aqueous solution and acetonitrile as mobile phase.The SB-C18 column(4.6 mm×250 mm,5μm)was used in HPLC,and C18 column with small packing particle size(2.1 mm×100 mm,1.5μm)was used in UPLC.The flow rates were 1.0 mL/min and 0.4 mL/min,respectively.The detection wavelength of lactucin and lactucopicrin were 258 nm,and that of chicoric acid,chlorogenic acid,esculin and aesculetin were 330 nm.The proposed methods have been successfully validated[relative standard deviation(RSD)<4%].The average recoveries ranged from 97.0%to 103.6%.The limits of detection(LODs)for the six components were from 0.49 mg/kg to 2.14 mg/kg in HPLC,and from 1.11 mg/kg to 5.24 mg/kg in UPLC.The limits of quantitation(LOQs)for the six components were from 1.65 mg/kg to 7.14 mg/kg in HPLC,and from 3.72 mg/kg to 17.50 mg/kg in UPLC.