Second slave voyage (1564–1565)
Hawkins persuaded Queen Elizabeth I to back his second slave voyage and secured investors from her court, these included Robert Dudley, 1st Earl of Leicester and Edward Clinton, 1st Earl of Lincoln, with William Cecil, 1st Baron Burghley involved as a supervisor. The queen allowed Hawkins to charter one of her ships for the trip, the 700 Ton Jesus of Lübeck and for it to fly her flag, the Royal Standard. He set sail from Plymouth on 18 October 1564 with four vessels, Jesus of Lübeck, and 3 ships of his own. He took over 400 people from Africa and enslaved them, some he bought from the Portuguese, others he kidnapped directly by raiding the coast, he left Africa on 29 January 1565.
Upon arriving at Borburata on April 3, 1565, Hawkins was informed that he could not trade without an authorised license from the governor. He requested permission to sell some of the ‘lean and sicke Negreos’ to help alleviate the burden on his ships and enable the sick to recover. After days of negotiations, local officials finally consented to purchase a few enslaved individuals while waiting for the governor's decision. He then travelled to Rio de la Hacha where he used force to ensure he secured the deals he thought fair. He sold 300 enslaved people, clothing, linen and wine. He was paid in gold, silver and other precious items and took further orders to sell enslaved people on a future voyage.
After completing his business, Hawkins prepared to return to England. Needing water, he sailed to the French colony of Fort Caroline in Florida. Finding them in need, he traded his smallest ship and a quantity of provisions to them for cannon, powder, and shot, that they no longer needed, as they were preparing to return to France. The provisions gained from Hawkins enabled the French to survive and prepare to move back home as soon as possible.
The voyage returned a profit reported at 60%. Hawkins brought back to England the sweet potato and tobacco, which was initially used as a narcotic; smoking did not gain in popularity until years later.