建立了固相萃取反相高效液相色谱荧光检测法测定拟南芥中生长素的方法。采用Waterssunfire C18(150mm×4.6mm,5μm)色谱柱,以乙腈-10mmol/L乙酸钠(20:80,V/V,乙酸调节至pH3.5)为流动相,在流速1mL/min,进样量20μL,柱温20℃,样品温度10℃,荧光激发和发射波长分别为275和345nm条件下,分离测定了拟南芥中的植物生长素吲哚乙酸和吲哚丙酸含量。分离效果良好,线性范围分别为85.71pg~21.43ng和0.33ng~1.29μg,相关系数分别为0.9992和0.9999,相对标准偏差小于5%,平均回收率分别为103.8%和102.6%。本方法可对少量模式植物样品进行灵敏、准确的定量分析。
An analytical method has been developed for the simultaneous determination of indole-3-acetic acid(IAA) and indole-3-propionic acid(IPA) in Arabidopsis thaliana by solid phase extraction and high performance liquid chromatography with fluorescence detection on a Waters sunfire C18(150 mm×4.6 mm,5 μm) chromatographic column.Acetonitrile and 10 mmol/L sodium acetate-acetic acid solution(pH 3.5)(20:80,V/V) was used as mobile phases at a flow rate of 1 mL/min.The column temperature was 20 ℃ and the temperature of sample was 10 ℃.The linear range of the IAA and IPA was 85.71 pg-21.43 ng and 0.33 ng-1.29 μg,the correlation coefficient was 0.9992 and 0.9999,the recoveries of IAA and IPA were 103.8% and 102.6%,respectively.The relative standard deviation(RSD,%) was less than 5%.This method can be used for determination of the auxin in Arabidopsis thaliana.