Çolak Nergiz Gürbüz, Eken Neslihan Tek, Frary Anne, Doğanlar Sami
Department of Molecular Biology and Genetics, Faculty of Science, Izmir Institute of Technology, Izmir, Turkey.
Plant Science and Technology and Application Center, Izmir Institute of Technology, Izmir, Turkey.
Bio Protoc. 2021 Mar 5;11(5):e3929. doi: 10.21769/BioProtoc.3929.
Targeted metabolomics is a useful approach to evaluate crop breeding studies. Antioxidant and flavor-related traits are of increasing interest and are considered quality traits in tomato breeding. The present study presents chromatographic methods to study antioxidants (carotenoids, vitamin C, vitamin E, phenolic compounds, and glutathione) and flavor-related characters (sugars and organic acids) in tomato. Two different extraction methods (for polar and apolar entities) were applied to isolate the targeted compounds. The extraction methods developed in this work were time and cost-effective since no further purification was needed. Carotenoids, vitamin C, glutathione, and phenolic acids were analyzed by HPLC-PDA using a RP C18 column at an appropriate wavelength for each compound. Vitamin E and sugars were analyzed by HPLC with RP C18 and NH2 columns and detected by FLD and RI detectors, respectively. In addition, organic acids were analyzed with GC-FID using a Rtx 5DA column after derivatization with MSTFA. As a result, sensitive analytical methods to quantify important plant metabolites were developed and are described herein. These methods are not only applicable in tomato but are also useful to characterize other species for flavor-related and antioxidant compounds. Thus, these protocols can be used to guide selection in crop breeding.
靶向代谢组学是评估作物育种研究的一种有用方法。抗氧化和风味相关性状越来越受到关注，并被视为番茄育种中的品质性状。本研究介绍了用于研究番茄中抗氧化剂（类胡萝卜素、维生素C、维生素E、酚类化合物和谷胱甘肽）和风味相关特征（糖类和有机酸）的色谱方法。应用了两种不同的提取方法（用于极性和非极性物质）来分离目标化合物。本研究中开发的提取方法既节省时间又经济高效，因为无需进一步纯化。类胡萝卜素、维生素C、谷胱甘肽和酚酸通过HPLC-PDA使用RP C18柱在每种化合物的适当波长下进行分析。维生素E和糖类分别通过使用RP C18柱和NH2柱的HPLC进行分析，并分别通过FLD和RI检测器进行检测。此外，有机酸在使用MSTFA衍生化后通过使用Rtx 5DA柱的GC-FID进行分析。结果，开发了用于定量重要植物代谢物的灵敏分析方法，并在此进行了描述。这些方法不仅适用于番茄，也有助于表征其他物种中的风味相关和抗氧化化合物。因此，这些方案可用于指导作物育种中的选择。
