As shown by many researches, various lipids are not only involved in many cellular processes, such as plant signal transduction, vesicular transport, and cytoskeletal rearrangements, but also play a pivotal role in mediating plant growth, development and stress responses. However, due to the diversity and complexity of lipids, and the limitation of analytical approach, its hard to have a deep understanding on lipids. Electrospray ionization tandem mass spectrometry (ESI-MS/MS) is a direct-infusion, high-throughput analytical strategy. By utilizing this approach, most lipid molecular species can be quantified in a short time only need small samples, detection of minor changes in plant lipids induced by environmental changes and growth and development has thus been greatly facilitated. The successful application of this approach in plants has led to great progress in plant lipidomics in recent years. Functions of some lipids and lipid-hydrolyzing enzymes in plant stress and defense responses have been revealed, and several novel genes related to plant lipid metabolism have been identified, all owing to the facilitation of this approach. Moreover, some new discoveries related to lipid distribution, transport, and transformation and identification of new lipid species have been made through the ESI-MS/MS approach coupled with other lipid analyzing methods. The paper firstly gives a brief depiction on the characteristics of the ESI-MS/MS approach, then provides an overview of application advances of the approach in plant lipidomics, and offers some prospective options to its future development in the final part.