高效液相色谱法分离和测定小麦中的5种内源激素  

Determination of five endogenous hormones in wheat by high performance liquid chromatography

作　　者：张玉琼[1] 仲延龙[1] 高翠云[1] 董召荣[2] 陈娜[1] 王梅方[1] 

机构地区：[1]安徽农业大学生命科学学院,安徽合肥230036 [2]安徽农业大学农学院,安徽合肥230036

出　　处：《色谱》2013年第8期800-803,共4页Chinese Journal of Chromatography

基　　金：农业部科技项目国家公益性行业(农业)科研专项(201103001;201203032;201203096);安徽省自然科学基金面上项目(1208085mc48)

摘　　要：建立了高效液相色谱法(HPLC)用于分离和测定小麦中的吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA3)、玉米素(ZT)和水杨酸(SA)5种植物内源激素。经过条件优化,选用甲醇作为样品提取溶剂。然后,经石油醚和乙酸乙酯萃取,经Sep-Pak C18小柱纯化。液相色谱的分离采用Eclipse XDB-C18(250 mm×4.6 mm,5μm)反相色谱柱;流速为1 mL/min;进样量10μL。检测器波长设置为254 nm;14.5 min时切换到240 nm;SA洗脱后即18 min时切换回254 nm。流动相A为甲醇,B为乙酸溶液(pH3.6)。梯度条件为0～7 min,20%A;7～10 min,20%A～28%A;10～17 min,28%A;17～19 min,28%A～40%A;19～35 min,40%A。结果表明,小麦中各激素的分离效果理想,加标回收率达96.9%～98%,相对标准偏差在1.54%～2.29%之间。因此,该方法的建立为快速、准确地分离和测定小麦内源激素提供了可靠的方法。A high performance liquid chromatographic （HPLC） method was developed to deter- mine the five endogenous hormones including indole-3-acetic acid （IAA）, abscisic acid （ ABA）, gibberellic acid （ GA3 ）, zeatin （ZT） and salicylic acid （SA） in wheat. The separation conditions were optimized, and methanol was chosen as the extraction solvent. Then the extract was extracted by petroleum ether and ethyl acetate, and purified with the Sep-Pak Cl8 column. The chromatographic conditions were as follows： Eclipse XDB-C,~ reversed phase col- umn （250 mm ×4.5 mm, 5 μm）, the flow rate of 1 mL/min, the injection volume of 10 μL. and the detection wavelength of 240 nm were used for the separation of SA from 14.5 min to 18 min, while the detection at 254 nm used for the separation of the others. Methanol （A） and acetic acid aqueous solution （ pH 3.6 ） （B） were used as the mobile phases with the linear gra- dient set as follows： 0-7 min20%A, 7-10 rain20% A-28%A, 10-17 min28%A, 17 -19 min 28% A -40% A, 19 -35 min 40% A. The results showed that： the hormones were separated well with the recoveries of 95.9% -98%, and the RSDs were in the range of 1.54% to 2.29%. It is a reliable method for rapid, accurate separation and determination of the endogenous hor- mones in wheat. Key words ：

关 键 词：高效液相色谱法 内源激素 小麦 

分 类 号：O658[理学—分析化学]