HPLC-DAD-FLD同时测定柑橘果汁中12种多甲氧基黄酮和香豆素类物质

摘要

建立高效液相色谱-串联二极管阵列检测器和荧光检测器(high performance liquid chromatography-diode array detector-fluorescence detector,HPLC-DAD-FLD)同时检测柑橘果汁中12种多甲氧基黄酮、香豆素及呋喃香豆素类物质的方法.以0.01％磷酸、甲醇、乙腈组成三元流动相进行梯度洗脱,12种物质在30 min内实现基线分离.利用DAD和FLD获得各物质的紫外和荧光光谱信息,将柑橘果汁样品组分的光谱与之比对,并结合色谱保留时间,实现样品成分的定性分析.分别以320 nm和450 nm为紫外和荧光的检测波长,研究该方法的定量性能.结果显示:标准曲线线性关系良好;荧光定量限低至μg/L级,可作为紫外定量的重要补充手段;果汁回收率的紫外和荧光检测值分别为95.2％～104.8％和94.5％～103.5％.该方法对样品定性、定量分析准确,适合柑橘多甲氧基黄酮、香豆素等能发射荧光物质的检测.%A method using high performance liquid chromatography with diode array detection and fluorescence detection (HPLC-DAD-FLD) was developed to simultaneously analyze 12 compounds of polymethoxyflavones (PMFs),coumarins and furocoumarins in citrus juice samples.Baseline separation of twelve compounds was achieved within 30 min using gradient elution with a ternary mobile phase composed of 0.01％ phosphoric acid,methanol and acetonitrile.Qualitative identification of the components in citrus juices was confirmed by comparing their UV and fluorescence (FL) spectra and retention times with those of the reference standards.Quantitative analysis of the juice components was operated at a UV wavelength of 320 nm (OD320 nm) and a fluorescence emission of 450 nm.The calibration curves showed good linearity.The limit of quantitation (LOQ) with FLD was at as low as μg/L level so that FLD could be an important complementary tool to UV detection.The recovery tests showed a good accuracy for both UV (95.2％-104.8％) and FL (94.5％-103.5％) examination.In general,this method is accurate and reliable for qualitative and quantitative analysis of citrus juice components,especially the ones having fluorescence emission like PMFs and coumarins.