The decision comes after the Catalan government insisted on imposing a 15-minute delay before passengers could be picked up.
Uber has been the target of protests by taxi drivers in Barcelona who say their business is being undermined.
A rival service, Spain's Cabify, has followed Uber's example.
Uber said the new restrictions left it with "no choice" but to suspend its services "while we assess our future in Barcelona".
Although ride-hailing has been suspended, the UberEats food delivery service will continue to operate in the city.