The drawing is notable for its complex composition, demonstrating the alternation in the positioning of figures that is first apparent in Leonardo's paintings in the Benois Madonna.

This turning posture is first indicated in paintings by Leonardo in the Adoration of the Magi and is explored in a number of drawings, in particular the various studies of the Virgin and Child with a cat that are in the British Museum.

Saint Anne smiles adoringly at her daughter Mary, perhaps indicating not only maternal pride but also the veneration due to the one who "all generations will call...blessed".

Although the older of the two children, John the Baptist humbly accepts the blessing, as one who would later say of his cousin "I am not worthy even to unloose his sandals".

[6] Saint Anne's hand, her index finger pointing toward Heaven, is positioned near the heads of the children, perhaps to indicate the original source of the blessing.

The composition differs from Leonardo's only other surviving treatment of the subject, The Virgin and Child with Saint Anne in the Louvre, in which the figure of the Baptist is not present.

The earliest reference to it is by the biographer Giorgio Vasari who, writing in the mid sixteenth century, says that the work was created while Leonardo was in Florence, as a guest of the Servite Monastery.

A date of 1498–99 is put on the work by Padre Sebastiano Resta, who wrote to Giovanni Pietro Bellori saying that Leonardo had drawn the cartoon in Milan at the request of Louis XII of France.

More recent historians have dated the work as early as the mid 1490s and, in the case of Carlo Pedretti and Kenneth Clark, as late as 1508–1510.

[10] Martin Kemp notes that the hydraulic engineering images appearing among the sketches in the preparatory drawing in the British Museum dates the composition to around 1507–1508, when Leonardo was making similar studies in the Codex Atlanticus.

[12] Amid fears that it would find an overseas buyer, it was exhibited in the National Gallery where it was seen by more than a quarter of a million people in a little more than four months, many of whom made donations in order to keep the artwork in the United Kingdom.

The shooter was identified as a mentally ill man by the name of Robert Cambridge, who claimed he committed this act in order to bring attention to "political, social and economic conditions in Britain".