HPLC同时测定黄檗种子中4种内源植物激素

Determination of Four Endogenous Phytohormones in Seeds of Phellodendron amurense by High Performance Liquid Chromatography (HPLC)

摘要: 本文以东北天然红松林重要伴生植物黄檗(Phellodendron amurense)种子为研究材料，采用高效液相色谱法对种子中的植物激素进行分析，以探讨植物种子中的植物激素测定方法。黄檗种子经80%甲醇浸提，C-18固相萃取柱萃取，反相C18柱分离，流动相：甲醇:乙腈:水 = 1:1:8 (体积比)。检测波长：GA为206 nm、IAA 225 nm、ABA与玉米素Z的检测波长为254 nm，首先在HPLC预设置206 nm (GA)和225 nm (IAA)两个波长，在样品运行7分钟时，手动更改检测波长为254 nm。利用该方法测定赤霉素、吲哚乙酸、玉米素和脱落酸4种内源植物激素的回收率在93%以上，相对标准偏差小于4.1%，精密度和重现性均较好。表明该方法适用于黄檗种子中植物激素含量的检测。

Abstract: Four endogenous phytohormones: Gibberellins, indoleacetic acid, zeatin and abscisic acid in the seeds of Phellodendron amurense, were examined by High Performance Liquid Chromatography (HPLC) to provide effective and reliable methods for identification of phytohormones in plant seeds. The four analytes were extracted from P. amurense seeds by 80% methanol and C-18 solid extraction column, and then separated by Agilent C18 column, using methanol, acetonitrile, water (1:1:8, V/V/V) as mobile phases. Detection wavelength: GA was 206 nm, IAA was 225 nm, ABA and Zeatin were 254 nm. First, the HPLC preseted 206 nm (GA) and 225 nm (IAA) as detection wavelengths, and when the sample ran for 7 minutes, the detection wavelength was changed to 254 nm. This method had been applied to the determination of gibberellin, indole acetic acid, zeatin and abscisic acid in the seeds of P. amurense, and the recovery rate of 4 kinds of endogenous phytohormones was more than 93%, relative standard deviation less than 4.1%. The results showed that phytohormones was well separated, and the ideal quantitative data of phytohormones in P. amurense seeds were obtained, and this method suited the purpose of the determination for phytohormones in P. amurense seeds.