提取工艺对色谱法检测芦荟叶多酚含量影响

Determination of Polyphenols in Aloe Leaf by HPLC Method and the Influence of Extraction Condition on Components' Contents

试验采用高效液相色谱法法比较不同提取工艺对芦荟叶中没食子酸(Gallic acid)、原儿茶酸(Protocate-chuicacid)、儿茶素(Catechin)、绿原酸(Chlorogenicacid)、表儿茶素(Catechin)、咖啡酸(Caffeicacid)、阿魏酸(Ferulicacid)和芦丁(Rutin)8种多酚含量的影响。试验采用ZORBAX SB-C18色谱柱(125 mm×4.6 mm×5μm),流动相A相为乙腈;流动相B相为0.1(V/V)甲酸,梯度洗脱,流速为0.8 mL/min,DAD检测波长为280 nm,柱温为45℃。检测结果表明,不同提取工艺得到的芦荟叶粉中没食子酸、原儿茶酸、儿茶素、绿原酸、表儿茶素、咖啡酸、阿魏酸和芦丁8种芦荟叶多酚的总含量以超临界二氧化碳萃取最高,超临界二氧化碳萃取过程没有加热处理,最大程度地保留了芦荟叶多酚;该检测方法在加标为1 000~100 000μg/kg浓度范围内的回收率为98.21%~103.28%,精密度为1.29%~4.67%(n=6),方法的检出限为0.02~0.05 mg/L,检测线性范围最宽为0.01~500 mg/kg;结果表明,从芦荟叶多酚的含量及物质特性保留的角度考虑,选择芦荟叶的提取工艺以超临界二氧化碳萃取为宜。

The influence of different extraction technologies on contents of eight polyphenols （gallic acid, protocatechuicacid, catechin, chlorogenic acid, catechin, caffeic acid, ferulicacid and rutin） from aloe leaf was compared with HPLC methodin this study. The experiment was based on ZORBAX SB-C15 column （125 mm x 4.6 mmx 5 μm） with mobile phase A ofacetonitrile and mobile phase B of 0.1 （V/V） formic acid, and gradient eluted with the flow rate of 0.8 mL/min, DAD detectionwavelength of 280 nm and column temperature of 45 ℃. It was shown by the results that the total content of 8 polyphenolsin aloe leaf （gallic acid, protocatechuic acid, catechin, chlorogenic acid, catechin, caffeic acid, ferulicacid and rutin） was thehighest when using supercritical extraction among all the extraction technologies. There was no heating treatment duringsubcritical extraction process, therefore, the polyphenols in aloe leaf could be kept at maximum. The recovery of this detectionmethod was 98.21%-103.28% in the spiked 1 000-100 000 btg/kg concentration, precision of 1.29%-4.67% （n=6）, limitof quantification was 0.02-0.05 mg/L, detection of linear range of 0.01-500 mg/kg. It was shown as well that, from the aspects ofincreasing polyphenols content and retaining materiales, it was the best to choose superccal extraction to exlract the aloe leaf.Koywords aloe leaf; chlorogenicacid; catechin; high-performance liquid chromatographic （HPLC）