基于超高效液相色谱同时测定木本植物中12种内源激素的方法及应用  

Simultaneous Determination of Twelve Endogenous Hormones in Woody Plants by Ultra Performance Liquid Chromatography and Its Application

作　　者：胡晓雯 黄海娇 张雯雯 许志茹 刘关君[1,2] HU Xiaowen;HUANG Haijiao;ZHANG Wenwen;XU Zhiru;LIU Guanjun(National Key Laboratory of Forest Genetics and Breeding,Haerbin150040;College of Forestry,Northeast Forestry University,Haerbin 150040;College of Life Science,Northeast Forestry University,Haerbin 150040)

机构地区：[1]林木遗传育种全国重点实验室,黑龙江哈尔滨150040 [2]东北林业大学林学院,黑龙江哈尔滨150040 [3]东北林业大学生命科学学院,黑龙江哈尔滨150040

出　　处：《分析科学学报》2024年第3期300-308,共9页Journal of Analytical Science

基　　金：黑龙江省自然基金联合引导项目(LH2022C015)。

摘　　要：建立了超高效液相色谱同时测定木本植物中吲哚乙腈(IAN)、吲哚丙酮酸(IPYA)、吲哚乙酰胺(IAM)、色胺(TAM)、吲哚-3-乙酸(IAA)、玉米素(ZT)、反式玉米素核苷(tZR)、脱落酸(ABA)和赤霉素A1(GA1)、赤霉素A3(GA3)、赤霉素A4(GA4)、赤霉素A7(GA7)等12种内源激素的分析方法。对前处理与测定条件进行了筛选和优化,木本植物组织经80%甲醇提取,通过固相萃取柱(MCX)纯化,以ZORBAX EP-C18为色谱柱,甲醇/0.1%冰乙酸为流动相梯度洗脱,柱温30℃,波长为254 nm和210 nm条件下,利用超高效液相色谱进行外标定量分析。测定12种激素线性相关系数R2>0.99,方法检出限(LOD)及定量限(LOQ)分别为0.002~0.029 ng·mL^(-1)和0.006~0.088 ng·mL^(-1)。样本84K杨和白桦在不同加标水平下的回收率分别为78.1%~116.8%和82.9%~114.2%,相对标准偏差(RSD)分别为0.3%~4.7%和0.2%~4.2%。应用本方法对两种木本植物组织内源激素含量的测定,结果表明,脱落酸含量最高分别为768.88 ng·g^(-1)、366.27 ng·g^(-1),GA3、GA4与GA7等活性赤霉素含量偏低。所建立的方法适用于杨树、桦树中多种激素的快速纯化分析,为其他木本植物激素的测定提供了借鉴。An ultra performance liquid chromatography method(UPLC)was developed for the simultaneous determination of twelve kinds of endogenous hormones(indole acetonitrile,indole pyruvate,indole acetamide,tryptamine,3-indoleacetic acid,zeatin,trans-zeatin riboside,abscisic acid,gibberellin A 1,gibberellin A 3,gibberellin A 4,and gibberellin A 7)in Woody plant.The woody plant tissues were extracted with 80%methanol.Purification of samples was performed with a MCX solid phase extraction column,the chromatography was performed on a ZORBAX EP-C18 column with gradient elution of methanol/0.1%glacial acetic acid at the column temperature of 30℃and detection wavelength of 254 nm and 210 nm.The limits of detection(LODs)and quantification(LOQs)were in the range of 0.002-0.029 ng·mL^(-1) and 0.006-0.088 ng·mL^(-1).The recoveries of each substance under different spiking levels were 78.1%-116.8%and 82.9%-114.2%,and the relative standard deviations(RSD)were 0.3%-4.7%and 0.2%-4.2%.The results showed that the content of abscisic acid was the highest(768.88 ng g^(-1) and 366.27 ng·g^(-1)),and the contents of GA3,GA4 and GA7 were lower.The established method could be used to purify and detect various hormones in 84K Poplar and Betula platyphylla Suk,and it provided a good reference for the determination of hormones in other woody plants.

关 键 词：前处理 超高效液相色谱 外标法 内源激素 木本植物 

分 类 号：O657.72[理学—分析化学]