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Analysis of Indole-3-Acetic Acid Metabolites from Dalbergia dolichopetala by High Performance Liquid Chromatography-Mass Spectrometry

机译:高效液相色谱-质谱法分析黄柏中的吲哚-3-乙酸代谢物

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摘要

A mixture of [2-14C1] and [13C6]indole-3-acetic acid was applied to the cotyledons of 6-day-germinated seeds of “jacarandá do cerrado” (Dalbergia dolichopetala) and after 8 hours the seeds were extracted. Analysis of the fractionated extract by reversed-phase high performance liquid chromatography-radiocounting revealed the presence of five radiolabeled metabolite peaks (I-V). After further purification, the individual peaks of radioactivity were analyzed by combined high performance liquid chromatography-steel filter-fast atom bombardment-mass spectrometry. The metabolite fraction V was found to contain [14C1, 13C6]indole-3-acetylas-partic acid and unlabeled indole-3-acetylglutamic acid. Analysis of the metabolite fraction II revealed the presence of dioxindole-3-acetylaspartic acid and putative dioxindole-3-acetylglutamic acid as well as putative benzene ring-hydroxylated derivatives of oxindole-3-acetylaspartic acid and oxindole-3-acetylglutamic acid. There was no evidence of significant incorporation of label from [2′-14C1] or [13C6]indole-3-acetic acid into any of these conjugated indoles.
机译:将[2- 14 C1]和[ 13 C6]吲哚-3-乙酸的混合物施用于“jacarandádo”的6天发芽种子的子叶中cerrado(Dalbergia dolichopetala),并在8小时后将种子提取出来。通过反相高效液相色谱-放射计数对分馏出的提取物进行分析,发现存在五个放射性标记的代谢物峰(I-V)。进一步纯化后,通过高效液相色谱-钢过滤器-快速原子轰击-质谱联用分析单个放射性峰。发现代谢物级分V含有[ 14 C1, 13 C6]吲哚-3-乙酰基-天冬氨酸和未标记的吲哚-3-乙酰基谷氨酸。代谢物级分II的分析显示存在二恶吲哚-3-乙酰基天冬氨酸和假定的二恶吲哚-3-乙酰基谷氨酸以及恶唑-3-乙酰基天冬氨酸和恶吲哚-3-乙酰谷氨酸的苯环羟基化衍生物。没有证据表明[2'- 14 C1]或[ 13 C6]吲哚-3-乙酸中的标记物显着掺入这些结合的吲哚中。

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