反相高效液相色谱法测定草莓天竺葵素-3-O-葡萄糖苷的含量

Determination of pelargonidin 3-O-glucoside in strawberry by reversed phase high performance liquid chromatography

为了测定草莓中天竺葵素-3-O-葡萄糖苷的含量,建立了一种反相高效液相色谱法。采用Waters Symmetry C18色谱柱（5μm,4.6 mm×150 mm）,流动相A为10%甲酸,流动相B为乙腈。线性梯度洗脱设计为：0～13 min,乙腈为0～20%,20 min,乙腈为40%,25 min,乙腈为0。检测波长为520 nm,进样量为每针20μL,柱温25℃,流速为1 mL·min^-1。结果表明,天竺葵素-3-O-葡萄糖苷在浓度为0.25～20μg·mL^-1范围内有良好的线性关系,线性回归方程为y=71 859x＋13 792,相关系数为0.9993,回收率为98.29%～102.69%,相对偏差3.03。本测定方法简便、准确、快速、稳定,适合于测定草莓及其他蔬菜水果中天竺葵素-3-O葡萄糖苷的含量。

In order to detect the content of pelargonidin 3-O-glucoside in strawberry,a new method using reversed phase HPLC was established. The HPLC system consisted of a Waters 1525 Binary Pump and a Waters 2487 dual λabsorbance detector under the control of Breeze software. The volume of 20 μL was injected onto a symmetry C18column（ 5 μm,4. 6 mm × 150 mm,Waters） with a symmetry C18security guard column（ 5 μm,3. 9 mm × 20 mm）at a flow rate of 1 mL·min^-1. The mobile phase was composed of 10% aqueous formic acid water（ A） and acetonitrile（ B）. A linear gradient was performed as follows： 0-20% B（ 0-13 min）,20%-40% B（ 13-20min）,40%-0 B（ 20-25 min）. The column temperature was set at 25℃ and the anthocyanin components were detected at 520 nm. The standard working curve had a good linear relationship in the concentration ranged from 0. 25-20 μg·mL^-1,the linear equation was y =71859x ＋13792 and the related coefficient was 0.993. The recoverieswere 98. 29%-102. 69% and the relatively standard deviation was 3. 03. This method is simple,fast,exact,steady and suitable to detect the content of pelargonidin 3-O-glucoside in strawberry and other plants.