高效液相色谱法分析黑米皮花青素及其体内代谢产物

Determination Anthocyanins from Black Rice Outlayer and Human Metabolites by HPLC

建立分析黑米皮中花青素含量及人体内代谢产物的高效液相色谱方法。并研究了黑米皮中花青素种类、含量及在人体内的代谢情况。样品经预处理后,以HypersilBDSC18柱,4%磷酸水溶液、乙腈(V/V)等度洗脱分离,520nm波长下检测、外标法定量。结果表明,黑米皮中花青素总含量约为2.31%,其中矢车菊素-3-葡萄糖苷(Cy-3-g)占1.87%,芍药素-3-葡萄糖苷(Pn-3-g)占0.44%。Cy-3-g在血浆中的最高浓度为21.47±4.48ng/ml,出现在口服黑米皮后1.5h,血浆中未检测到Pn-3-g。Cy-3-g在0～2h的尿液样品中的含量最高(390.78±69.28ng/ml),Pn-3-g在2～4h收集的尿液中含量最高(105.90±26.26ng/ml)。

The determination of the anthocyanins content in black rice outlayer and the human metabolites were studied. After samples preparation, the anthocyanins were determined with an UV-HPLC system at 520nm detection. A G18-column was used at a column temperature of 30℃. The mobile phase, 4% aqueous H3PO4/CH3CN （8.8：1.2 V/V）, was used under isocratic conditions at a flow rate of 0.8ml/min. The results showed that total anthocyanin contents of black rice outlayer fraction were 2.31%. Cyanidin 3-glucoside accounted for 1.87% of anthocyanin contents and Peonidin 3-glucoside accounted for 0.44% of anthocya- nin contents. The plasma Cy-3-gmax value was 21.47 ±4.48ng/ml at 1.5h after intaking the black rice outlayer fraction and Pn- 3-g can＇ t be detected in plasma sample. The urine Cy-3-gmax, and Pn-3-gmax values were 390.78 ± 69.28ng/ml 2h after intake and 105.90± 26.26ng/ml 4h after intake, respectively.