同时测定鲜切荸荠黄化组织中柚皮素和圣草酚的高效液相色谱条件探究

Exploration of simultaneous determination for naringenin and eriodictyol in etiolation tissues of fresh- cut Chinese water- chestnut

采用Agilent Eclipse XDB-C18色谱柱,通过考察流动相、柱温、流速等色谱条件建立了能同时测定鲜切荸荠黄化组织中柚皮素和圣草酚的高效液相色谱法。色谱条件为柱温:40℃;流动相:0.5%冰醋酸溶液(A)-乙腈(B),梯度洗脱(0~10 min,10%→20%B;10~15 min,20%→30%B;15~25 min,30%→40%B;25~35 min,40%→50%B);流速:1 m L·min^(-1);进样量:10μL;检测波长:280 nm。此色谱条件下,柚皮素和圣草酚均能得到有效分离,分别在5.60~28.00μg·m L^(-1)(r=1.0000)、3.20~16.00μg·m L^(-1)(r=0.9999)范围内峰面积与浓度的线性关系良好,加标回收率分别在99.49%~100.71%、98.68%~101.65%(n=6)。本研究所得的高效液相色谱条件简便快捷、准确可靠,重现性好,能排除其中所含其他物质色谱和其他物质产生的干扰,因此可用于同时测定鲜切荸荠黄化组织中的柚皮素及圣草酚。

To explore the simultaneous determination of naringenin and eriodictyol in etiolation extract of fresh-cut Chinese water-chestnut, HPLC was established by optimizing the chromatographic conditions including mobile phase,column temperature, flow rate, etc. in using Agilent Eclipse XDB-C18 column. The optimal simultaneous determination conditions were the mobile phase composed of 0.5% （v/v） acetic acid in water （eluent A） and acetonitrile（eluent B）in a linear gradient program of 0-10 min,lO%-20% B, 10-15 min of 20%-30% B,15-25 min of 30%-40% B and 25-35 min of 40%-50% B at a continual flow rate of 1.0 mL. min t at a column temperature of 40℃,the injection volume was 10 uL,and the UV detection was performed at 280 nm,which could make naringenin and eriodictyol separate successfully. Naringenin and eriodictyol revealed high correlation between 5.60- 28.00 ug·mL J （ r = 1.0000） and between 3.20-16.00 ug· mL-1 （ r = 0.9999） and exhibited recovery range of 99.49 % 100.71% and 98.68% - 101.65% （n = 6）, respectively.The chromatographic conditions were simple, accurate and reliable,with good reproducibility, which can eliminate the chromatography of other substances contained and interference of other substances produced, so it can be used as a method that simultaneously determine naringein and eriodictyol in etiolation tissues of fresh-cut Chinese water-chestnut.