Beyond the wall: High-throughput quantification of plant soluble and cell-wall bound phenolics by liquid chromatography tandem mass spectrometry
Beyond the wall: High-throughput quantification of plant soluble and cell-wall bound phenolics by liquid chromatography tandem mass spectrometry
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DOI：
10.1016/j.chroma.2018.12.059
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发表时间：
2019-03-29
影响因子：
4.1
通讯作者：
Alonso, Ana Paula
中科院分区：
文献类型：
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作者：
Cocuron, Jean-Christophe;Casas, Maria Isabel;Alonso, Ana Paula
植物体内积累了数千种酚类化合物，包括木质素和黄酮类化合物，这些化合物主要通过苯丙烷途径合成，在植物的生长和适应中起着重要作用。建立了一种新的高通量超高效液相色谱串联质谱（UHPLC-MS/MS）方法，用于定量测定19种黄酮类化合物和15种其他酚类化合物，包括酸、醛和醇。在10分钟内进行色谱分离，允许拆分异构体，例如3-、4-和5-绿原酸、4-羟基苯甲酸和水杨酸、异Orientin和Orientin以及毛地黄黄酮和山奈酚。发现每种化合物的线性范围为低fmol至高pmol。此外，这种UHPLC-MS/MS方法显示出非常灵敏，检测限在1.5阿莫尔至300 fmol之间，定量限在5阿莫尔至1000 fmol之间。玉米幼苗提取物用于评估方法在回收率、基质效应和准确度方面的稳健性。在研究的34种代谢物中，生物基质未抑制32种代谢物的信号。此外，大多数分析物从生物样品中回收，效率高于75%。所有黄酮类化合物和其他酚类化合物的日内和日间准确度在+/- 20%的范围内，除了松柏醇和香草酸。最后，定量黄酮类化合物，游离和细胞壁结合酚类物质在幼苗中的两个玉米品系与对比酚含量成功地实现了使用这种方法。由爱思唯尔公司出版
Plants accumulate several thousand of phenolic compounds, including lignins and flavonoids, which are mainly synthesized through the phenylpropanoid pathway, and play important roles in plant growth and adaptation. A novel high-throughput ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) method was established to quantify the levels of 19 flavonoids and 15 other phenolic compounds, including acids, aldehydes, and alcohols. The chromatographic separation was performed in 10 min, allowing for the resolution of isomers such as 3-, 4-, and 5-chlorogenic acids, 4-hydroxybenzoic and salicylic acids, isoorientin and orientin, and luteolin and kaempferol. The linearity range for each compound was found to be in the low fmol to the high pmol. Furthermore, this UHPLC-MS/MS approach was shown to be very sensitive with limits of detection between 1.5 amol to 300 fmol, and limits of quantification between 5 amol to 1000 fmol. Extracts from maize seedlings were used to assess the robustness of the method in terms of recovery efficiency, matrix effect, and accuracy. The biological matrix did not suppress the signal for 32 out of the 34 metabolites under investigation. Additionally, the majority of the analytes were recovered from the biological samples with an efficiency above 75%. All flavonoids and other phenolic compounds had an intra- and inter-day accuracy within a +/- 20% range, except for coniferyl alcohol and vanillic acid. Finally, the quantification of flavonoids, free and cell wall-bound phenolics in seedlings from two maize lines with contrasting phenolic content was successfully achieved using this methodology. Published by Elsevier B.V.