Analysis of pesticide residues is very important to enforce legislation and guarantee food safety. The correct use of pesticides is still crucial in agriculture because they provide spectacular increases in crop yields and ensure global demand for grain. However, the indiscriminate, incorrect and/or excessive use of pesticides in agriculture may have some serious adverse effects such as the accumulation of residues in food. Pesticide residues are controlled worldwide by maximal residues limits (MRLs), not the same in all countries but generally ranging from a few μg kg-1 (usually for pesticides that are banned) to a few tens of mg kg-1. Determining pesticides at this concentration requires sensitive, accurate and robust instrumentation, and trained personnel as well. This chapter explores the latest advances to determine pesticide residues as accurately as possible in the shortest time. A description of aspects like improvement of high-throughput methods specificity and advances in the determination by gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) or (bio)sensors, are presented in this chapter. The focus is on multi-residue o multiplexed analysis that will offer rapidity and economy in order to achieve the required sensitivity (<0.01 mg kg-1). The primary purpose of this chapter is to provide the reader with a state- of- the- art assessment and identification of gaps within this field, and to establish future trends in the extraction, purification, and determination of pesticide residues.