UPLC-MS/MS分析不同品种蓝莓中的花青苷

Analysis of Anthocyanins in Different Blueberry Cultivars by UPLC-MS/MS

目的：采用超高效液相色谱-三重四级杆串联质谱联用技术（UPLC-MS/MS）分析12个品种蓝莓中的花青苷。方法：80%乙醇作为溶剂超声辅助提取蓝莓花青苷,提取液经离心后的上清液于旋转蒸发仪浓缩用6%甲酸水溶液定容至刻度线,超滤后,采用UPLC-MS/MS,ACQUITY UPLC BEHC18色谱柱（2.1 mm×50 mm,1.7μm）,乙腈-6%甲酸水溶液作为流动相,进行梯度洗脱,在电喷雾离子源（ESI）正离子模式下,用MRM模式监测。结果：12个品种蓝莓中花青苷种类较为相似,花青苷种类最丰富的蓝莓品种是R26,含有23种,花青苷种类最少的蓝莓品种是N15,含有20种,飞燕草素和锦葵素是蓝莓中最主要的两大类花青苷。12个品种蓝莓中总花青苷含量在820.43-2 114.627μg·g-1,存在显著差异（PN12〉S28〉R6〉S30〉S7-1〉S12〉S7-2〉S7-3〉N15〉N1〉N14。R系列总花青苷含量高于S系列,N系列花青苷含量最少。不同品种蓝莓中各类花青苷含量亦存在显著差异。结论：12个品种蓝莓中的总花青苷含量存在显著差异,可为以蓝莓花青苷为功能成分的保健品、药物开发提供基础数据,同时也为蓝莓花青苷活性实验、蓝莓花青苷的检测分析提供参考依据。

Objective： To analyze the anthocyanins in 12 varieties of blueberries by ultra performance liquid chromatography-tandem triple-quadrupole mass spectrometry（ UPLC-MS/MS）. Method： The 80% ethanol was used as solvent to extract anthocyanin from blueberry under assistance of ultrasound,and the centrifugated supernatant of the extract was treated in a rotating evaporator and concentrated in 6% formic acid solution to the scale line. After ultrafiltration,the extract anthocyanin was analyzed on ACQUITY UPLC BEH C18 column（ 2. 1 mm × 50 mm,1. 7 μm）,with 6% formic acid and acetonitrile as the mobile phase for gradient elution. The positive ion mode of electrospray ionization（ ESI） was adopted in the course of multiple reactions monitoring（ MRM）. Result： The anthocyanins of 12 varieties were similar. R26 blueberry had the most abundant anthocyanin species（ 23 species）. N15 blueberry had the least anthocyanin species（ 20 species）. Delphinidin and malvidin were the two main types of anthocyanins in blueberries. The total anthocyanins content was 820. 43-2 114. 627 μg·g-1 in 12 varieties,with significant differences,R26〉 N12 〉 S28 〉R6 〉 S30〉 S7-1〉 S12 〉S7-2〉 S7-3〉 N15〉 N1 N14 from high to low. The total anthocyanins content in blueberrys of R series was higher than that in blueberrys of S series. The anothcyanins content in blueberrys of N series was lowest. There were also significant differences in contents of anthocyanins between different varieties of blueberries. Conclusion： The content of total anthocyanins in the 12 varieties of blueberries detected in the experiment showed significant differences,which could provide basic data for the development of health products and drugs with blueberry anthocyanin as a functional component,and also provide reference for the activity test of blueberry anthocyanin as well as the detection and analysis of blueberry anthocyanin.