The new rules state that passengers who choose to take a suitcase into the cabin with them will have pay £5 for priority boarding or have it put in the hold.
But flyers have said it is just a way for the the low-cost airline to ‘loot’ its passengers.
The Spanish Association of Users, Employers and Professionals of Air Transport (ASETRA) has called for passengers to boycott the airline in regards to the changes.
Passengers had been allowed to take on a small bag measuring 35cm x 20cm x 20cm and a suitcase no more than 55cm x 40cm x 20cm.
But now the larger suitcase must go into the hold unless paying for priority boarding.
ASETRA has urged Spain to give airline slots to ‘kinder’ companies in protest.
Passengers could also be charged €50 if they fail to adhere to the new rules.
ASETRA said: “This new condition violates the rights of passengers at a time when Ryanair enjoys a dominant position in the Spanish air transport market.
“This prevents users from choosing other airlines in some routes that the Ryanair company exclusively carries out and thus limits their ability to choose.”
ASETRA has stated that the new Ryanair rules could be challenged in the European courts due to a ‘change of contract in the agreed conditions.’
This applies to passengers who bought their ticket before the changes were announced last year in September.
Ryanair’s Kenny Jacobs told the Express.co.uk: “Since we announced this new policy, we have had a positive reception from our customers.
“This new policy is fair, will speed up boarding and will eliminate any risk of Ryanair flights being delayed because of too many bags being brought on board.
“These new bag rules are centred around lower checked bag fees and bigger bag allowances, changes which will cost Ryanair up to €50m p.a. so we hope our customers will enjoy and welcome them.
“All Ryanair customers can still bring 2 free carry-on bags but because of our heavily booked flights (94 per cent load factors), we don’t have space on board for this many wheelie bags so are asking non-priority customers to put their bigger bag in the hold – free of charge – from today in order to eliminate boarding delays and improve our industry leading on-time departures.”