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通过气相萃取和气相色谱-化学电离质谱联用同时定量分析植物中的茉莉酸和水杨酸。

Simultaneous quantification of jasmonic acid and salicylic acid in plants by vapor-phase extraction and gas chromatography-chemical ionization-mass spectrometry.

作者信息

Engelberth Juergen, Schmelz Eric A, Alborn Hans T, Cardoza Yasmin J, Huang Juan, Tumlinson James H

机构信息

Center for Medical, Agricultural and Veterinary Entomology, United States Department of Agriculture, Agricultural Research Service, 1700 SW 23rd Drive, Gainesville, FL 32608, USA.

出版信息

Anal Biochem. 2003 Jan 15;312(2):242-50. doi: 10.1016/s0003-2697(02)00466-9.

Abstract

Jasmonic acid and salicylic acid represent important signaling compounds in plant defensive responses against other organisms. Here, we present a new method for the easy, sensitive, and reproducible quantification of both compounds by vapor-phase extraction and gas chromatography-positive ion chemical ionization-mass spectrometry. The method is based on a one-step extraction, phase partitioning, methylation with HCl/methanol, and collection of methylated and, thus, volatilized compounds on Super Q filters, thereby omitting further purification steps. Eluted samples are analyzed and quantified by GC/MS with chemical ionization. Standard curves were linear over a range of 5-1000 ng for jasmonic acid and salicylic acid. The correlation coefficients were greater than 0.999 and the recovery rates estimated between 70 and 90% for salicylic acid and 90 and 100% for jasmonic acid. The limit of detection was about 500 fg by using single ion detection mode. Both, cis- and trans-isomers for jasmonic acid can be detected. A comparison with established methods indicates the new method to be highly efficient, allowing reliable quantification of both compounds from small amounts of plant material (5-400mg fresh weight).

摘要

茉莉酸和水杨酸是植物对其他生物防御反应中的重要信号化合物。在此,我们提出了一种新方法,可通过气相萃取和气相色谱 - 正离子化学电离 - 质谱法轻松、灵敏且可重复地定量这两种化合物。该方法基于一步萃取、相分配、用盐酸/甲醇进行甲基化,以及在Super Q滤纸上收集甲基化从而挥发的化合物,因此省略了进一步的纯化步骤。洗脱后的样品通过化学电离气相色谱/质谱进行分析和定量。茉莉酸和水杨酸的标准曲线在5 - 1000 ng范围内呈线性。相关系数大于0.999,水杨酸的回收率估计在70%至90%之间,茉莉酸的回收率在90%至100%之间。使用单离子检测模式时,检测限约为500 fg。茉莉酸的顺式和反式异构体均可被检测到。与现有方法的比较表明,该新方法效率极高,能够从小量植物材料(5 - 400 mg鲜重)中可靠地定量这两种化合物。

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