采用液质联用方法检测黄秋葵荚果黄酮类物质含量  

Content determination of flavonoids from the fruit seed of Abelmoschus esculentus by UPLC-QqQ-MS

作　　者：谢进[1] 黄艳宁[1] 徐瑞[1] 曹亮[1] 范海珊[1] 朱校奇[1] XIE Jin;HUANG Yan-Ning;XU Rui;CAO Liang;FAN Hai-Shan;ZHU Xiao-Qi(Institute of Agro Environment and Ecology, Hunan Academy of Agricultural Sciences, Changsha 410125, China)

机构地区：[1]湖南省农业科学院农业环境生态研究所,长沙410125

出　　处：《广西植物》2017年第12期1592-1597,共6页Guihaia

基　　金：湖南省自然科学基金(14JJ6071);国家公益性行业(农业)科研专项(201303117);湖南省农业科学院科技创新项目(2016QN01)~~

摘　　要：采用超声提取方法,利用液质联用仪建立快速的黄秋葵黄酮类物质提取检测方法,使其能应用于黄秋葵相关产品的品质检测。选定黄芩素、柚皮素、槲皮素、黄芩苷和芦丁这五种黄酮类物质,确定液相色谱、质谱条件和检测线性范围等,从而确定黄秋葵荚果黄酮类物质的提取检测方法。色谱条件:色谱柱为华谱unitary C18(2.1 mm×150 mm,5μm);流动相:A为乙腈,B为含0.1%甲酸的水;洗脱梯度:80%B^60%B(0~4 min),60%B(4~10 min),60%B^55%B(10~12 min),5%B(12.1~18 min),5%B^80%B(18~20min),80%B(20.1~25 min)。质谱条件:毛细管电压为4 000 V,气体温度为330℃,气体流速为12 L·min^(-1),喷雾器压力为275.8 k Pa。优化得到黄芩素、柚皮素、槲皮素、黄芩苷和芦丁这五个标样的裂解能量为34、22、30、18、34 eV;五个标样在11 min内完全分离,线性范围为0.5~400μg·L^(-1),线性相关系数均大于0.99。黄秋葵荚果采用超声波法提取黄酮(75%的乙醇,1∶10的料液比,超声75 min),通过实验得到整个提取检测过程中黄芩素、柚皮素、槲皮素、黄芩苷和芦丁的回收率分别为(93.65±3.66)%、(98.79±2.04)%、(100.66±1.83)%、(87.23±6.61)%、(101.91±0.61)%,并对黄秋葵果荚中这5种黄酮物质的含量进行了测定。该方法重复性好、灵敏度高、专一性强,为黄秋葵黄酮类物质的研究提供了实验基础。The ultra-performance liquid chromatography-triple quadrupole mass spectrometry（ UPLC-QqQ-MS） was applied to establish the rapid detection method for Abelmoschus esculentus flavonoids. This method can apply to qualify the products of A. esculentus. Rutin, baicalin, quercetin, baicalein, naringenin were selected as the standard compounds. The chromatographic separation was done on a ACCHROM UNITARY C18（ 2.1 mm × 150 mm,5 μm） in gradient elution with a mobile phase of acetonitrile（ A） and formic acid aqueous solution（ B）（ 0.1%,W/W） as follows： 80%B-60%B（ 0-4 min）,60%B（ 4-10 min）,60%B-55%B（ 10-12 min）,5%B（ 12.1-18 min）,5%B-80%B（ 18-20 min）,80%B（ 20.1-25 min）. The mass spectrometry condition： The capillary voltage was 4 000 V. The gas temperature was 330 ℃. The gas velocity was 12 L·min-1. The spray pressure was 275.8 k Pa. The MS parameters were set up as： collision energy,34 eV for baicalein,22 eV for naringenin,30 eV for quercerin,18 eV for baicalin and 34 eV for rutin. The results showed good linearity for five flavonoids with R2 values ranging from 0.998 4-0.999 8. Extraction of A. esculentus was prepared by Ultrasonic method with 75% ethanol,1 ∶ 10 ratio of material to liquid,ultrasonic75 min. The recoveries rang from 87.23% to 101.91%（ RSD varied from 0.61% to 6.61%）. The content of these flavonoids in fruit seed of A. esculentus were detected. The method is rapid and has high sensitivity,high accuracy and good specificity. It provides the basic experimental method for the flavonoids of A. esculentus.

关 键 词：超高效液相色谱—三重四级杆质谱联用 黄秋葵荚果 黄酮类物质 

分 类 号：Q946[生物学—植物学]