The Spanish collection spans five centuries. The earliest work is a mid-fifteenth-century panel by Nicolás Francés. There are sixteenth-century paintings by Juan Fernández de Navarrete, Luis de Morales and by El Greco, who was active in Toledo.
The collection is particularly rich in works of the seventeenth century, the Golden Age of Spanish painting. Bartolomé Esteban Murillo and Francisco de Zurbarán, both of whom lived out most of their careers in Seville, are richly represented. Diego Velázquez moved from Seville to Madrid in 1623 where he was appointed Pintor del Rey. The collection holds The Supper at Emmaus, a painting from his early career and part of the Beit gift of seventeen masterpieces which were presented to the Gallery in 1987. The contribution of Jusepe de Ribera, who spent his mature career in Italy where he came under the influence of Caravaggio, is reflected in a number of religious subjects. There are also compositions by Juan van der Hamen y León, Pedro Núñez del Valle, José Antolínez and Juan Antonio de Frías y Escalante, and a still life by Juan Fernández (‘El Labrador’). The Gallery holds four canvases by Francisco José de Goya, among them his exquisite Portrait of Doña Antonia Zárate, which is part of the Beit gift.