LaBioMMi, Federal University of São Carlos, São Carlos, SP, Brazil; Max Planck Institute for Chemical Ecology, Jena, Germany; King Abdullah University of Science and Technology, Thuwal, Saudi Arabia.
Accert Chemistry and Biotechnology Inc., São Carlos, SP, Brazil.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Dec 1;1186:123006. doi: 10.1016/j.jchromb.2021.123006. Epub 2021 Oct 22.
Oxylipins constitute a huge class of compounds produced by oxidation of long-chain unsaturated fatty acids either chemically (by radicals such as reactive oxygen species, ROS) or enzymatically (by lipoxygenases, LOX; cyclooxygenases, COX; or cytochrome P450 pathways). This process generates fatty acids peroxides, which can then be further modified in a broad range to epoxy, hydroxy, keto, ether fatty acids, and also hydrolyzed to generate small aldehydes and alcohols. In general, oxylipins are present in almost all living organisms and have a wide range of signaling, metabolic, physiological, and ecological roles depending on the particular organism and on their structure. In plants, oxylipins have been extensively studied over the past 35 years. However, these studies have focused mainly on the jasmonates and so-called green leaves volatiles. The function of early LOX products (like keto and hydroxy fatty acids) is yet not well understood in plants, where they are mainly analyzed by indirect methods or by GC-MS what requires a laborious sample preparation. Here, we developed and validated a straightforward, precise, accurate, and sensitive method for quantifying oxylipins in plant tissues using HPLC-MS/MS, with a one-step extraction procedure using low amount of plant tissues. We successfully applied this method to quantify the oxylipins in different plant species and Arabidopsis thaliana plants treated with various biotic and abiotic stress conditions.
氧化脂类是一大类化合物，由长链不饱和脂肪酸的氧化产生，这种氧化可以通过化学途径（如活性氧物种 ROS）或酶促途径（如脂氧合酶 LOX、环氧化酶 COX 或细胞色素 P450 途径）实现。这个过程会生成脂肪酸过氧化物，然后可以进一步广泛修饰为环氧化物、羟基、酮基、醚脂肪酸，也可以水解生成小醛和醇。总的来说，氧化脂类几乎存在于所有生物中，根据特定的生物体和其结构，具有广泛的信号转导、代谢、生理和生态作用。在植物中，氧化脂类在过去 35 年中得到了广泛的研究。然而，这些研究主要集中在茉莉酸及其所谓的绿叶挥发物上。在植物中，早期 LOX 产物（如酮基和羟基脂肪酸）的功能尚未得到很好的理解，它们主要通过间接方法或 GC-MS 进行分析，这需要繁琐的样品制备。在这里，我们开发并验证了一种使用 HPLC-MS/MS 直接、精确、准确和灵敏地定量植物组织中氧化脂类的方法，该方法采用一步提取程序，只需少量植物组织。我们成功地将该方法应用于定量不同植物物种和各种生物和非生物胁迫条件下处理的拟南芥植物中的氧化脂类。
