固相萃取-液相色谱-串联质谱法同时测定蜂蜜中16种黄酮类化合物和阿魏酸

Simultaneous determination of 16 flavonoids and ferulic acid in honey by solid phase extraction and high performance liquid chromatography-tandem mass spectrometry

建立了固相萃取净化-液相色谱-串联质谱法(SPE-LC-MS/MS)同时测定蜂蜜中芦丁、杨梅素、桑黄素、槲皮素、柚皮素、橙皮素、木犀草素、染料木素、山柰酚、异鼠李素、芹菜素、松属素、汉黄芩素、白杨素、高良姜素、芫花素和阿魏酸含量的方法。蜂蜜经pH 2的盐酸溶液稀释,C18固相萃取柱净化,液相色谱-串联质谱法检测,外标法定量。以空白蜂蜜基质溶液配制0~200μg/kg的系列标准溶液,线性相关系数大于0.997,方法定量限为20μg/kg。在蜂蜜样品中进行加标水平为20、40、100μg/kg的添加回收试验,回收率为64.5%~113%,相对标准偏差为1.4%~14.5%。该方法取样量少、操作简便、快捷,可用于蜂蜜中黄酮类化合物的测定。

Flavonoids are polyphenol compounds that are widely distributed in honey;they play an important role in its antioxidant activity and aid in distinguishing various species of honey.An analytical method based on solid phase extraction and high performance liquid chromatography with tandem mass spectrometry(SPE-HPLC-MS/MS)for the simultaneous determination of rutin,myricetin,morin,quercetin,naringenin,hesperetin,luteolin,genistein,kaempferol,isorhamnetin,apigenin,pinocembrin,wogonin,chrysin,galangin,genkwanin,and ferulic acid in honey was developed and validated.The honey samples were diluted with HCl solution(pH 2)and cleaned up on a C18 column by solid phase extraction;the methanol eluent was filtered through a nylon membrane and analyzed by LC-MS/MS on an ODS-3 column(150 mm×4.6 mm,3μm).Quantitative detection was performed by LC-MS/MS in the multiple reaction monitoring(MRM)mode under negative electrospray ionization(ESI-)conditions.The external standard method was used for quantitation.The method validation data were obtained.The calibration standard curve solutions were prepared by honey matrix-matching;the linear range was 0-200μg/kg,with correlation coefficients of the calibration standard curves greater than 0.997.The limits of quantification(LOQs)of this method were 20μg/kg.The recoveries were in the range of 64.5%-113%,and the relative standard deviations were 1.4%-14.5%at three spiked levels(20,40,and 100μg/kg).The developed method is convenient,rapid,and effective,and it is suitable for the determination and confirmation of flavonoids in honey samples.