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.
Cabify, which has one million users in Barcelona, said the city had "given in to the demands of the taxi sector, seriously hurting citizens' interests".
It said the move would put 3,000 jobs at risk in the city.
In July last year, Spanish taxi drivers began public protests in Barcelona and Madrid.
The drivers are still on strike in the Spanish capital, but Madrid authorities have said they will not adopt the same restrictions as in Barcelona.
Uber has fallen foul of restrictions in many parts of the world at different times.
In the UK, Transport for London stripped it of its licence in late 2017, saying it was not a "fit and proper" operator.
It was then awarded a short-term 15-month licence for London in June last year, with London mayor Sadiq Khan saying the firm had been "put on probation".