Pulses stand out in the dietary realm due to their rich reservoir of both macro- and micronutrients. Beyond their nutritional benefits, these foods also harbor a complex array of compounds known as antinutrients (ANs). This group of ANs, which includes proteins, glycosides, tannins, saponins, and alkaloids, has the potential to hinder the gastrointestinal absorption of essential nutrients, thereby introducing possible health concerns. Particularly salient antinutrients found in pulses are phytic acid (PA), lectins, saponins, and various enzyme inhibitors, notably trypsin, chymotrypsin, and α-amylase inhibitors. This chapter meticulously explores the historical progression of methodologies designed to evaluate the levels of phytic acid, lectins, and saponins in pulses. Highlighting the advancements achieved over the last 50 years, the contemporary techniques highlighted herein not only boast improved sensitivity and accuracy but also facilitate meticulous quantification. This precision paves the way for comprehensive comparative analyses, especially relevant when examining plant-based proteins derived from pulses.