高效液相色谱法在小麦激素测定中的优化  

Optimization of Hormone Determination of Wheat by High Performance Liquid Chromatography

作　　者：唐晓东[1] 蒋春志[1] 徐俊杰[1] 张丽[1] 樊翠芹[1] 于翠红[1] TANG Xiao-dong;JIANG Chun-zhi;XU Jun-jie;ZHANG Li;FAN Cui-qin;YU Cui-hong(Institute of Cereal and Oil Crops,Hebei Academy of Agriculture and Forestry Sciences/Crop Genetics and Breeding Laboratory of Hebei Province,Shijiazhuang 050035,China)

机构地区：[1]河北省农林科学院粮油作物研究所/河北省作物遗传育种实验室,河北石家庄050035

出　　处：《河北农业科学》2021年第3期97-103,108,共8页Journal of Hebei Agricultural Sciences

摘　　要：针对现有液相色谱检测植物激素方法检测效率低的问题,利用本试验建立了高效液相色谱法测定小麦组织中反式玉米素(tZ)、反式玉米素核苷(tZR)、赤霉素A3(GA3)、6-苄氨基嘌呤(6-BA)、吲哚-3-乙酸(IAA)、脱落酸(ABA)和α-萘乙酸(NAA) 7种激素的方法,并将单个样品检测时间缩短到15 min。本方法采用双pH萃取法提取激素,以ZORBAX Eclipse XDB-C18色谱柱分离,甲醇-0.8%冰醋酸梯度洗脱,检测波长260 nm,流速0.8 mL/min,柱温40℃。该试验条件下,能实现7种激素有效分离。7组标准曲线线性关系良好,R2范围为0.999 7~1.000 0。tZ、tZR、GA3、6-BA、IAA、ABA、NAA的加标回收率分别为91.45%±0.29%、97.96%±0.77%、96.65%±1.17%、 93.40%±0.45%、104.21%±1.85%、92.61%±0.15%和96.65%±2.84%。该方法检测激素种类多、检测时间短、重复性良好、适用性强,极大地提高了高效液相色谱法在现有激素检测中的检测能力和检测效率。In view of the low detection efficiency of the existing liquid chromatography methods for plant hor mone,a high performance liquid chromatography(HPLC) method was established for the determination of trans zeatin(t Z),trans zeatin nucleoside(t ZR),gibberellin A3(GA3),6-Benzylaminopurine(6-BA),indole-3-acetic acid(IAA),abscisic acid(ABA) and α-naphthylacetic acid(NAA) in wheat tissues. The detection time was shortened to 15 min. The hormone was extracted by double p H extraction, separated on Zorbax Eclipse XDB-C18 column, eluted with methanol-0.8% glacial acetic acid gradient, the detection wavelength was260 nm,the flow rate was 0.8 mL/min,and the column temperature was 40 ℃. Under the experimental condi tions,7 kinds of hormones can be separated effectively. The linear relationship of the standard curves of 7 groups was good,and R2 was 0.999 7-1.000 0. The recovery rates of t Z, t ZR, GA3,6-BA,IAA,ABA and NAA were 91.45%±0.29%,97.96%±0.77%,96.65%±1.17%,93.40%±0.45%,104.21%±1.85%,92.61%±0.15% and96.65%±2.84%,respectively. This method could determine many kinds of hormones with short detection time,and had good repeatability and strong applicability,which greatly improved the detection ability and efficiency of HPLC in the existing hormone detection.

关 键 词：小麦 激素 高效液相色谱 双pH萃取 

分 类 号：S512.1[农业科学—作物学]