Shiva Sunitha, Enninful Regina, Roth Mary R, Tamura Pamela, Jagadish Krishna, Welti Ruth
1Kansas Lipidomics Research Center, Division of Biology, Kansas State University, Manhattan, KS USA.
2Department of Agronomy, Kansas State University, Manhattan, KS USA.
Plant Methods. 2018 Feb 13;14:14. doi: 10.1186/s13007-018-0282-y. eCollection 2018.
Lipidomics plays an important role in understanding plant adaptation to different stresses and improving our knowledge of the genes underlying lipid metabolism. Lipidomics involves lipid extraction, sample preparation, mass spectrometry analysis, and data interpretation. One of the practical challenges for large-scale lipidomics studies on plant leaves is the requirement of an efficient and rapid extraction method.
A single-extraction method with a polar solvent mixture gives results comparable to a widely used, multi-extraction method when tested on both and leaf tissue. This single-extraction method uses a mixture of 30 parts chloroform, 25 parts isopropanol, 41.5 parts methanol, and 3.5 parts water (v/v/v/v) and a 24-h extraction time. Neither inclusion of ammonium acetate nor inclusion of acetic acid increased extraction efficiency.
The extract produced by this method can be used for analysis by mass spectrometry without a solvent evaporation step. The amount of lipid extracted, including phosphatidic acid, is comparable to widely used, more labor-intensive methods. The single-extraction protocol is less laborious, reducing the potential for human error.
脂质组学在理解植物对不同胁迫的适应性以及增进我们对脂质代谢相关基因的了解方面发挥着重要作用。脂质组学包括脂质提取、样品制备、质谱分析和数据解读。在对植物叶片进行大规模脂质组学研究时，一个实际挑战是需要一种高效快速的提取方法。
当在[具体植物1]和[具体植物2]叶片组织上进行测试时，一种使用极性溶剂混合物的单提取方法所得到的结果与一种广泛使用的多提取方法相当。这种单提取方法使用由30份氯仿、25份异丙醇、41.5份甲醇和3.5份水（体积比/体积比/体积比/体积比）组成的混合物，提取时间为24小时。加入醋酸铵或醋酸均未提高提取效率。
该方法产生的提取物无需进行溶剂蒸发步骤即可用于质谱分析。所提取的脂质量，包括磷脂酸，与广泛使用的、更耗费人力的方法相当。单提取方案操作更简便，减少了人为误差的可能性。
