Under the ban, live races and simulcast betting on dog races will be banned in the state - which is considered the hub of the country's dog-racing industry with 11 tracks. This will leave only six operational tracks in the US.
The new legislation, which is set to come into place by 2020, won the vote by 69-31.
'Save thousands of dogs'
"Shutting down the tracks will save thousands of gentle greyhounds from being caged for 22 to 23 hours a day, given performance-enhancing drugs - including cocaine - and forced to run to the point that many sustain broken legs, develop heatstroke, or suffer heart attacks," said vegan charity PETA.
"Dogs used for racing have incurred spinal injuries, broken necks, and crushed skulls, and some were electrocuted when they collided with electric fences.
"On average, one dog dies every three days on a Florida racetrack. Now, all greyhound racing will end in Florida by 2020."
The bipartisan legislation was introduced by Florida State Senator Tom Lee, and advocated by GREY2K USA Worldwide, the Doris Day Animal League, the ASPCA, and the Humane Society of the United States among others.