高效液相色谱法测定玉米内源激素方法的改进研究

Study on the Improvement of HPLC Method for Analyzing Endogenous Hormone in Maize

以玉米植株为材料,对提取、纯化和利用高效液相色谱法分析测定玉米植株中内源激素生长素(IAA)、脱落酸(ABA)、赤霉素(GA3)和玉米素(ZT)的含量的方法进行改进。针对玉米植株的糖分、叶绿素含量高等特点,对以往的HPLC分析植物内源激素的植物材料前处理方法进行改进。试材用液氮研磨以80%预冷的甲醇浸提,浸提液利用旋转蒸发仪浓缩、石油醚去除色素、乙酸乙酯提取激素、PVP去除酚类杂质,最后用0.45 um微孔滤膜抽滤,紫外检测器在254 nm波长下检测吸收波长,以甲醇和水位流动相,利用高效液相色谱法检测内源激素含量,回收率分别为IAA 93.79%±7.43%,ABA 95.79%±6.14%,GA388.63%±6.31%,ZT 89.69%±7.34%。

This experiment improved the method of extraction, purification and analysis by high-performance liquid chromatography（HPLC） on endogenous hormones auxin（IAA）, abscisic acid（ABA） and gibberellic acid（GA3） and zeatin（ZT） content in maize plant tissues. To deal with the existence of large amount of sugar and chlorophyll. The test materials were grinded in liquid nitrogen and lixiviated in 80% methanol, then pigment was eliminated in petroleum ether, phytohormones were extracted in ethyl acetate, phenolic was compounded in polyving pyrrolidone（PVP） and quantified by 0.45 urn microporous membrane filter. Detection was made by UV detector at 254 nm. Reversed HPLC with methanol-water at a flow rate of 1.0 mL/min. The frequency of reclamation was IAA 93.79% ±7.43%, ABA 95.79% ± 6.14%, GA3 88.63% ± 6.31%, ZT 89.69% ± 7.34%.