ZHANG Hai jun,GUO Li,WANG Ming ze,et al.Determination of Total Flavonoids Content in Arabidopsis thaliana by High Performance Liquid Chromatographic[J].HEILONGJIANG AGRICULTURAL SCIENCES,2014,(04):97-100.
HPLC法测定拟南芥中总黄酮含量的研究
- Title:
- Determination of Total Flavonoids Content in Arabidopsis thaliana by High Performance Liquid Chromatographic
- 文章编号:
- 1002-2767（2014）04-0097-04
- Keywords:
- Arabidopsis thaliana; total flavonoids content; rutin; highperformance liquid chromatographic(HPLC)
- 分类号:
- R284.2
- 文献标志码:
- A
- 摘要:
- 为更好地开发和利用拟南芥中总黄酮及相关副产品,研究了检测拟南芥中总黄酮含量的方法。结果表明:以芦丁为标准品，建立了检测拟南芥中总黄酮含量的HPLC法。HPLC法色谱条件：C18色谱柱、35℃柱温、256 nm检测波长、35%甲醇〖DK1〗∶65%水（含0.4%磷酸）流动相、10 μL进样量及10 mL?min-1流速。标准品芦丁在该色谱条件下，峰型良好，在0.05～1.60 mg?mL-1时峰面积与质量浓度间呈良好线性关系，R2=0.999 7，平均加样回收率为99.90%，RSD小于3%(n=6)，该方法适用于拟南芥中总黄酮的检测。
- Abstract:
- In order to develop and use the flavonoids and related byproducts in Arabidopsis thaliana,the determine method was established for total flavonoids content.The results showed that taking rutin as the standard,highperformance liquid chromatographic method was established,mobile phase consisted on methanol(A)/0.4% orthophosphoric acid in distrilled water(B)and each solvent was filtered through a 0.45 μm pore size filter and degassed in an ultrasonic bath before.The analysis of rutin was performed at 35℃(column oven temperature)with 10 mL?min-1 flow rate,the injection volume was 10 μL,and solvent A∶B of rutin was 35%∶65%,the chromatogram was detected at 256 nm.The peak area had a good linearity with concentration when R2=0.999 7,the recoveries of standard spiling was 99.90% with RSD less than 3%.So the method was suitable for determination of total flavonoids in Arabidopsis thaliana.
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备注/Memo
收稿日期：2013-12-27基金项目：教育部新教师基金资助项目（3M2101936604）；国家麻类产业技术体系资助项目（NYCYTX-19-S13）；黑龙江省农业科技创新工程重点（青年基金）资助项目第一作者简介：张海军（1986-），男，山东省济宁市人，硕士，研究实习员，从事植物分子育种和技术推广工作。E-mail：zhanghaijun_234@163.com。通讯作者：王殿奎（1963-），男，黑龙江省双城市人，研究员，从事亚麻、大麻、菊芋等经济、能源作物的新品种选育、产业开发和技术推广工作。E-mail：359351633@qq.com。