首页|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

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

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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.
Simultaneous Determination of Twelve Endogenous Hormones in Woody Plants by Ultra Performance Liquid Chromatography and Its Application
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 A1,gibberellin A3,gibberellin A4,and gibberellin A7)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.

PretreatmentUltra performance liquid chromatographyExternal standard methodEndogenous hormoneWoody plant

胡晓雯、黄海娇、张雯雯、许志茹、刘关君

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林木遗传育种全国重点实验室,黑龙江哈尔滨 150040

东北林业大学林学院,黑龙江哈尔滨 150040

东北林业大学生命科学学院,黑龙江哈尔滨 150040

前处理 超高效液相色谱 外标法 内源激素 木本植物

黑龙江省自然基金联合引导项目

LH2022C015

2024

分析科学学报
武汉大学,北京大学,南京大学

分析科学学报

CSTPCD北大核心
影响因子:0.717
ISSN:1006-6144
年,卷(期):2024.40(3)
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