高效液相色谱法同时检测棉花根中的多种植物激素

目的：探寻高效液相色谱同时检测棉花根中多种植物激素含量的方法。方法：采用WatersC18反相色谱柱（4.6×250mm,5μm）,在柱温为35℃、流速为1mL.min-1的条件下,以乙腈和三乙胺溶液为流动相梯度洗脱,在每种物质的保留时间附近切换至最大吸收峰（GA3除外）波长作为检测波长,并与254nm同一波长检测多种植物激素含量的方法进行比较,分离和检测棉花根中的玉米素（Z）、玉米素核苷（ZR）、赤霉酸（GA3）、生长素（IAA）和脱落酸（ABA）的含量。结果：切换波长法检测5种植物激素的灵敏度和回收率均较高,检出限均较低。回收率为：Z 96.82%、ZR 94.14%、GA3 92.75%、IAA 93.38%、ABA 95.57%;检出限为：Z 0.1μg.mL-1、ZR 0.1μg.mL-1、GA3 0.5μg.mL-1、IAA 0.3μg.mL-1;ABA 0.05μg.mL-1,能准确检测出棉花根中Z、ZR、GA3、IAA和ABA的含量。结论：采用Waters C18反相色谱柱（4.6×250mm,5μm）,在柱温为35℃、流速为1mL.min-1的条件下,以乙腈和三乙胺溶液为流动相梯度洗脱,结合切换波长法能同时检测出植物组织中多种植物激素含量。

Simultaneous Determination of Several Phytohormones in Cotton Root by using High Performance Liquid Chromatography

Objective： To explore how to simultaneously detect the content of several phytohormones in cotton root by high performance liquid chromatography （HPLC）. Methods： The contents of zeatin（z）, ribosylzeatin（ZR）, gibberellin（GA3）, growth hormone（IAA）, abscisin （ABA）were detected by the following conditions： reversed phase column Waters C 18（4.6× 250mm, 5 μm） of 35 ℃, gradient elution by using acetonitrile and triethylamine solution as mobile phase with flow speed 1mL·min^-1, the detection wavelength for every substance denpending on it＇s maximum absorptive summit wavelength during the retension time. Results： Sensitive and recovery rate all were higher, recovery rate of five phytohormones as Z 96.82%, ZR 94.14%, GA3 92.75%, IAA 93.38%, ABA 95.57%.Detection limit of five phytohormones all were lower, Z 0.1 μg· mL^-1, ZR 0.1μg· mL^-1, GA3 0.5 μg·mL^-1, IAA 0.3 μg·m^-1, ABA 0.05 μg·mL^-1. The content of five phytohormones（Z, ZR ,GA3, IAA and ABA）can accuratly be detected. Conclusion： The study established the method which could simultaneously be detected the content of several phytohormones of plant tissue by high performance liquid chromatography （HPLC） through the wavelength change.