目的 建立白花蛇舌草中多成分同时定量的高效液相色谱法方法。方法 采用Ultimate XB C18(4.6 mm×250 mm, 5 μm)色谱柱, 以乙腈(B)-水(A)(各含体积分数为0.1‰乙酸)为流动相进行梯度洗脱, 流速0.8 mL·min-1, 柱温35 ℃, 检测波长238 nm;运用外标法和一测多评法分别测定白花蛇舌草中4个环烯醚萜类和2个黄酮类成分的含量。结果 白花蛇舌草中的车叶草酸, quercetin-3-O--β-D-glucopyranoside, kaempferol-3-O--β-D-galactopyranoside, (E)-6-O-香豆酰鸡屎藤次苷甲酯, (E)-6-O-阿魏酰鸡屎藤次苷甲酯, (Z)-6-O-香豆酰鸡屎藤次苷甲酯分别在2.34~93.50, 2.61~104.33, 0.67~26.69, 3.42~136.84, 0.65~26.07, 1.10~44.17 μg·mL-1内线性关系良好(r>0.999 3), 仪器精密度、方法重复性、样品稳定性的RSD值均不大于2.2%, 平均加样回收率(n=6)均在99.8%~101.1%, RSD均不大于1.2%, 外标法实测值与一测多评法计算值经配对t检验得P＞0.05。结论 两种定量方法的含量分析结果之间无显著性差异, 均可用于白花蛇舌草中上述4个环烯醚萜类和2个黄酮类成分的同时定量分析。
Abstract
OBJECTIVE To develop an HPLC METHOD for simultaneous determination of multiple-components in Hedyotis diffusa Willd. METHODS The HPLC analysis was carried out on a C18 column(4.6 mm×250 mm, 5 μm) by gradient elution with acetonitrile-water as mobile phase at a flow rate of 0.8 mL·min-1, the column temperature at 35 ℃, and the detection wavelength was set at 238 nm. External standard METHOD and quantitative analysis of multi-components by single marker(QAMS) METHOD were adopted for simultaneous determination of six components in Hedyotis diffusa Willd, respectively. RESULTS The linear ranges for asperulosidic acid, quercetin-3-O--β-D-glucopyranoside, kaempferol-3-O--β-D-galactopyranoside, (E)-6-O-p-coumaroyl scandoside methyl ester, (E)-6-O-feruloyl scandoside methyl ester, (Z)-6-O-p-coumaroyl scandoside methyl ester were 2.34-93.50, 2.61-104.33, 0.67-26.69, 3.42-136.84, 0.65-26.07, and 1.10-44.17 μg·mL-1(r>0.999 3), respectively. The RSD values of precision, reproducibility, and sample stability were not more than 2.2%. The average recoveries of the six components were 99.8%-101.1%with RSDs not more than 1.2%. The P values of external standard METHOD and QAMS by paired t-test were greater than 0.05. CONCLUSION There is no significant difference in the content analysis RESULTS of the two methods, which can both used for simultaneous determination of the four iridoids and two flavonoids in Hedyotis diffusa Willd.
关键词
白花蛇舌草 /
高效液相色谱法 /
环烯醚萜 /
黄酮 /
外标法 /
一测多评法 /
含量测定
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Key words
Hedyotis diffusa Willd /
HPLC /
iridoid /
flavonoid /
external standard method(ESM) /
quantitative analysis of multi-components by single marker(QAMS) /
content assay
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中图分类号：
R917
R284
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参考文献
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脚注
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基金
广东省自然科学基金研究团队项目(351063201000003)
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