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基质辅助激光解吸/电离飞行时间质谱法监测拟南芥叶片提取物中的花青素。

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry monitoring of anthocyanins in extracts from Arabidopsis thaliana leaves.

作者信息

Marczak Lukasz, Kachlicki Piotr, Koźniewski Piotr, Skirycz Aleksandra, Krajewski Paweł, Stobiecki Maciej

机构信息

Institute of Bioorganic Chemistry PAS, Noskowskiego 12/14, 61-704 Poznań, Poland.

出版信息

Rapid Commun Mass Spectrom. 2008 Dec;22(23):3949-56. doi: 10.1002/rcm.3819.

Abstract

Anthocyanins are secondary plant metabolites ubiquitous in the plant kingdom. They have different biological activities, so monitoring their content in plant tissue or in feed/food derived from plants may be an important task in different projects from various fields of molecular biology and biotechnology. Profiling of secondary metabolites with high-performance liquid chromatography/mass spectrometry (HPLC/MS) systems is time-consuming, especially when many samples have to be checked within a defined time frame with a reasonable number of repetitions according to the metabolomic standards. Even application of the advanced ultra-performance liquid chromatography (UPLC)/MS or equivalent systems would require a long time for analysis of numerous samples. We demonstrate the applicability of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for the assessment of level (concentration) of anthocyanins in leaf tissues of four Arabidopsis thaliana ecotypes grown at normal (20 degrees C/16 degrees C day/night) and decreased (4 degrees C) temperature. The quantitative results were obtained for anthocyanins with MALDI-TOF MS using ferulic acid as a matrix. The amounts of anthocyanins in leaves of A. thaliana varied from 0.3-2.5 microg per gram of leaves for ecotypes Col-0 and C24, respectively, and contents of these markedly increased in plants grown in the cold. The applied analytical method exhibited better repeatability of measurements than obtained with an HPLC/ion trap MS system.

摘要

花青素是植物界普遍存在的次生植物代谢产物。它们具有不同的生物活性,因此监测其在植物组织中或源自植物的饲料/食物中的含量,可能是分子生物学和生物技术各个领域不同项目中的一项重要任务。使用高效液相色谱/质谱(HPLC/MS)系统对次生代谢产物进行分析耗时较长,尤其是当必须在规定时间内按照代谢组学标准对大量样品进行合理次数的重复检测时。即使应用先进的超高效液相色谱(UPLC)/MS或等效系统,分析大量样品也需要很长时间。我们展示了基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)在评估四种拟南芥生态型叶片组织中花青素水平(浓度)方面的适用性,这些生态型在正常温度(20摄氏度/16摄氏度,白天/夜晚)和低温(4摄氏度)条件下生长。使用阿魏酸作为基质,通过MALDI-TOF MS获得了花青素的定量结果。对于生态型Col-0和C24,拟南芥叶片中的花青素含量分别为每克叶片0.3 - 2.5微克,并且在低温下生长的植株中这些花青素的含量显著增加。所应用的分析方法比HPLC/离子阱MS系统具有更好的数据重复性。

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