This almost life-size double-portrait of the daughters of Phillip II of Spain was painted when the Infantas unexpectedly became heirs to the huge Spanish Empire following the sudden deaths of their half-brother Don Carlos and their mother, Isabel de Valois in 1568. It is strikingly similar to Alonso Sánchez Coello's double-portrait of the infantas in the Museo del Prado, Madrid. The painting was formerly attributed to Anthonis Mor van Dashorst and subsequently to Sofonisba Anguissola; however, the overall composition and the modelling of the features and treatment of the hands points toward the work of Coello.

Coello was one of the foremost portraitists of the later sixteenth century and a pioneer of the Spanish tradition of portraiture. A follower of Titian, he combined the golden tones and use of light on the canvas found in Venetian painting, with a precision of representation learned in the studio of his Flemish Master, Anthonis Mor (RCIN 405799). Mor was court painter to Phillip II and on his return to the Netherlands in 1561, Coello took his former Master's place. Coello remained at the Spanish court in Madrid until his death in 1588 and continued to enrich the style inherited from both Titian and Mor, bringing a distinctive use of colour, sharpness of execution and heightened realism to Spanish court portraiture. This painting is very close in quality to other known works by Coello. However it lacks the fluency of Coello's paintwork, suggesting it was at least in part painted by one of the artists working in his studio. 

The painting is typical of sixteenth century Spanish court portraiture, depicting the little Infantas in formal poses and with a precise attention to detail in the painting of their richly ornamented dresses and jewellery. Five year old Isabella Clara Eugenia (1566-1633) is shown on the left, holding a green parrot on her hand. Across the table Infanta Catalina Michaela (1567-97) holds gloves in her left hand and rests her right on the forepaw of a spaniel who sits on the table. Both girls are wearing dark green dresses which are richly embroidered with flowers and gold lace and trimmed with pearls. Their ornate jewellery would have been a family heirloom and the collar and girdle worn by the Infantas in this portrait appear again in a portrait of their mother in the Prado (Pantoja de la Cruz, c. 1605). During this period it was common for royal children to be depicted almost as miniature adults and in this portrait, the 'al forza', a tuck of fabric around the hem of a skirt, which was a distinctive feature of dress for adult woman in Spain, is clearly shown.

In the Prado painting there are noticeable differences in the position of the children's hands and the objects with which they are playing – wreaths of flowers instead of pet animals. These differences indicate that artists working in Coello's studio were allowed some flexibility in their copies of the Master's paintings, although his treatment of the faces is minutely reproduced.

The painting appears in Pyne's illustrated 'Royal Residences' of 1819, hanging in the Presence Chamber at Kensington Palace (RCIN 922150).