高效液相色谱法测定不同开花时期大马士革Ⅲ玫瑰(Rosa damascene tyigintipetala)花青素种类及含量

本实验采用HPLC法对大马士革Ⅲ玫瑰(Rosa damascene tyigintipetala)蕾期、初开期、盛开期和盛开末期4个开花时期中6种主要花青素(飞燕草色素、矢车菊色素、天竺葵色素、芍药素、锦葵色素、矮牵牛色素)同时进行测定。采用V(无水乙醇)∶V(水)∶V(盐酸)=2∶1∶1溶液超声提取样品中花青苷30 min,并于沸水浴中水解1 h。测定的色谱条件为Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm),流动相为1%甲酸水溶液和1%甲酸乙腈溶液,流速0.8 mL/min,检测波长530 nm,柱温35℃。结果表明:大马士革Ⅲ玫瑰4个开花时期中均检出了矢车菊色素和天竺葵色素,方差分析结果表明两类花青素不同开花时期含量差异达到极显著水平(p<0.01);两类花青素含量均随开花过程和花瓣褪色而降低;矢车菊色素含量在不同开花时期均占绝对优势。同时,本研究建立的大马士Ⅲ玫瑰花青素的提取和检测方法,该方法简便、快速、稳定且重现性好,可作为大马士革Ⅲ玫瑰及其他植物源性材料中飞燕草色素、矢车菊色素、天竺葵色素、芍药素、锦葵色素、矮牵牛色素等六种花青素的有效测定方法。

Dertermination of Anthocyanidin Composition and Content from Rosa damascene tyigintipetala at Different Flowering Periods by HPLC

In this experiment,the HPLC method was used to determine 6 main anthocyanidins,including delphinidin pigment,cyanidin pigment,pelargonidi pigment,peonidin pigment,malvidin pigment and petunidin pigment,from Rosa damascene tyigintipetala at 4 different flowering periods(i.e.,unfold-petal stage,initial-flowering stage,fullflowering stage and flowering-wilting stage).The anthocyanidins in the sample were extracted by ultrasonication for 30 min with the extracting solvent(ethanol:water:hydrochloric acid=2:1:1),and hydrolyzed for 1 h in a boiling water bath.The HPLC was carried out by using an Agilent ZORBAX SB-C18 column(4.6 mm×250 mm,5μm),the flow phase was 1%formic acid aqueous solution and 1%formic acid acetonitrile solution,the flow rate was 0.8 mL·min^?1,the detection wavelength was 530 nm,and the column temperature was 35℃.The results showed that cyanidin and pelargonidin pigments were detected at 4 flowering periods from Rosa damascene tyigintipetala.Variance analysis showed that the content difference between these two anthocyanidins at different flowering stages was extremely significant(P<0.01).The content of these two anthocyanidins decreased with the flowering process and petal fading,and the content of cyanidin pigment was absolutely dominant at different flowering stages.The established extraction and detection method of anthocyanidins from Rosa damascene tyigintipetala in this study is simple,rapid,stable and reproducible,which can be further used as an effective method for the determination of 6 anthocyanidins,such as delphinidin pigment,cyanidin pigment,pelargonidi pigment,peonidin pigment,malvidin pigment and petunidin pigment from Rosa damascene tyigintipetala and other plant-derived materials.