The sitter is shown half-length, standing slightly to the right, in a black doublet and ruff with only his left hand visible. This holds the bottom of a sash with a silver-beaded edge, which is tied round his neck and threaded through a signet ring bearing unclear initials. The pommel of a sword worn across his back is visible under his right arm and his left wrist wears a discreet bracelet of pearls and (possibly) jet beads.

Hawkins was the first English slave trader and a highly successful merchant in other areas of trade. He made four voyages to Sierra Leone between 1564 and 1569, taking a total of 1200 Africans across the Atlantic to sell to the Spanish settlers in the Caribbean island of Hispaniola. On his first voyage he described capturing 300 Africans 'by the sword and partly by other means'. In the Caribbean he sold them to the Spanish for 'hides, ginger, sugars, and some quantity of pearls, but he freighted also two other hulks with hides and like commodities.'

In 1567, on his last slaving voyage, in which his younger kinsman Francis Drake accompanied him, all but two of his ships and some treasure were seized by the Spanish in the port of San Juan de Uloa, Mexico, though he escaped and was later able to recover some of his captured men from Spain. In 1572 he became MP for Plymouth and in 1577 Treasurer of the Navy, in which role he greatly improved construction of ships. He was a senior commander in the Armada campaign, during which he was knighted, and subsequently joint commander with Frobisher in an expedition against the Portuguese coast in 1590. After the Armada he set up the 'chest at Chatham', the Naval welfare fund for seamen and in 1592 the hospital (alms-house) which still bears his name there. Hawkins died in Puerto Rico, shortly before Drake, on their final expedition to the West Indies in 1595–96.

This portrait is inscribed 'Anno D[omi]ni 1581' , 'Aetatis Suae 44' and (possibly later) 'Sr John Hawkins', though if the age stated is correct it would have been painted in 1576. It is presumed to have been in the Hawkins Hospital at Chatham practically since its foundation, from whose Governors it was purchased for the NMM.