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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%。
Maize plants were used as materials for the determination of endogenous hormones IAA, ABA, GA3 and ZT in maize plants by extraction, purification and high performance liquid chromatography (HPLC) Content method to be improved. In view of the characteristics of maize plants such as high sugar content and chlorophyll content, the methods for the pretreatment of plant material with endogenous hormones by HPLC were improved. The samples were extracted with 80% precooled methanol by liquid nitrogen grinding. The extract was concentrated by rotary evaporator, the petroleum ether was used to remove the pigment, the ethyl acetate was used to extract the hormones and the PVP was used to remove the phenolic impurities. Finally, a 0.45 um microporous membrane The absorption wavelength was detected by UV-Vis detector and UV detector at 254 nm. The mobile phase was methanol and water. The content of endogenous hormones was detected by HPLC. The recoveries were 93.79% ± 7.43% for IAA and 95.79% ± 6.14 for ABA, respectively %, GA388.63% ± 6.31%, ZT 89.69% ± 7.34%.