English: Portrait of a Woman, inscribed in gold over red "Anna Bollein Queen". Black and coloured chalks on pink prepared paper, 28.1 × 19.2 cm. Royal Collection, Windsor.

The question whether or not this Holbein drawing is of Anne Boleyn has produced sharp division among scholars. It comes from a priceless collection discovered by Queen Caroline of Ansbach in a bureau at Kensington in 1727. The drawing is one of two by Holbein inscribed as of Anne Boleyn, the other being in the British Museum. The attribution of the present drawing as Anne Boleyn was made by John Cheke, tutor of the future Edward VI, in 1542. Cheke had entered the royal service after a period working at Cambridge University, and so he may never himself have seen Anne Boleyn, who had been executed in 1536; however, he knew people who had known her. For a long time, many scholars, including K. T. Parker and Anne Boleyn's biographer Eric Ives, have doubted that this portrait was of Anne: they point to the occasional mistake in Cheke's attributions, to the sketches of Wyatt heraldry on the back of the sheet, and to dissimilarities between this image and other possible likenesses of Anne, who was said to have had darker hair than depicted. It has also been argued that Holbein would not have drawn a woman of Anne's stature in an undercap. In Ives's view, "There is little to reinstate either Holbein drawing". Among those who argue the case for this portrait being correctly inscribed are Holbein scholar John Rowlands, historian David Starkey, and Holbein's biographer Derek Wilson. They argue for the reinstatement of this image as of Anne and express reluctance to dismiss Cheke's attribution. Rowlands challenges Ives's conclusions, which are partly based on dissimilarities with other possible images of Anne, on the grounds that it is a mistake to rely too much on the accuracy of these other images, particularly since none, except for a damaged portrait medal, are provably contemporary with Anne. Rowlands concludes that "the circumstantial grounds in favour of the Windsor drawing are really very compelling".

As a result of these disagreements, the drawing has not been captioned consistently in reproductions, sometimes being called "Anne Boleyn" without reservation, and sometimes "Unknown Lady" or something similar. Some scholars prefer to label the drawing less decisively: Susan Foister, for example, the curator of the Tate's "Holbein in England" exhibition of 2006, writes: "Whether Holbein portrayed Anne remains an open question: a drawing at Windsor inscribed with her name shows a fair-haired woman whose appearance differs greatly from the painted portraits"; Tarnya Cooper, in the catalogue of the "Elizabeth" exhibition at the National Maritime Museum in 2003, captions the drawing: "Portrait of a Woman, probably Anne Boleyn, c. 1532–6".