GC--MS内标法定量分析植物组织中的JA、 IAA和ABA
Quantitative analysis of jasmonic acids, indole-3-acetic acid and abscisic acid in plant tissues by GC-MS
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摘要: 提出了利用气相色谱--质谱联用仪同时定量测定植物组织中茉莉酸 (JA)、吲哚--3--乙酸 (IAA) 和脱落酸 (ABA) 的方法。实验以欧洲灰杨叶片为材料, 在用80%冷甲醇提取激素时, 一并加入ABA、 IAA和JA的内标(D3 --ABA 300 ng,13C6 --IAA 60 ng 和DHJA 80 ng), 激素提取液浓缩至水相, 调pH2.5~3.0后经乙酸乙酯萃取、Sep--Pak C18小柱纯化, 甲酯化后, GC--MS定量测定ABA、JA。IAA经甲酯化后再经TMS化 (三甲基硅烷化), 用GC--MS定量测定。经检测: 盆栽欧洲灰杨叶组织中脱落酸含量为120.170~128.771 ng/g(鲜质量), 茉莉酸含量为1.697~6.974 ng/g(鲜质量), 吲哚乙酸含量为4.990~14.776 ng/g(鲜质量); 脱落酸含量最高, 其次是吲哚乙酸, 茉莉酸含量最低。Abstract: GC-MS was used to determine the concentrations of jasmonic acid (JA), indole-3-acetic acid (IAA) and abscisic acid (ABA) in Populus canescens leaves in this paper. Abundant cold methanol (80%, v/v) was used to extract JA, IAA and ABA, and three internal standards, D3-ABA 300 ng,13C6-IAA 60 ng and DHJA 80 ng were simultaneously introduced into the leaf samples. Crude extract was condensed by vacuum evaporating and hormones were re-extracted by ethyl acetate at pH 2.5--3.0 (×3). Then condensed JA, IAA and ABA extracts were purified by Sep-Pak C18 columns. Thereafter, partially purified hormones were methylated using trimethylsilyldizomethane and then analyzed by GC-MS (JA and ABA). IAA was analyzed by GC-MS after an additional TMS reaction. Results showed that concentrations of ABA, JA, and IAA in leaf tissues of P. canescens were 121.064--128.771 ng/g, 2.190--6.974 ng/g and 4.990--15.012 ng/g Fw, respectively. In concentration in P. canescens leaves, ABA IAA JA.
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Keywords:
- GC-MS /
- internal standard method /
- jasmonic acid /
- indole-3-acetic acid /
- abscisic acid /
- Populus canescens
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期刊类型引用(10)
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