Manchester United moves up from fourth to second on the list, which is based on season 2013-14 revenues.
Bayern Munich, Barcelona and Paris Saint-Germain filled the next spots.
English clubs Manchester City, Chelsea, Arsenal and Liverpool were placed sixth to ninth, all of them showing healthy rises in revenues.
Tottenham were in 13th spot, while there were new entries for Newcastle United and Everton at 19th and 20th.
The total combined revenue for the top 20 richest clubs rose by 14% to €6.2bn, the report found. The list only looks at revenues accrued and does not take into account club debts.
Real Madrid, which won its so-called "Decima" European Cup/Champions League after beating city neighbours Atletico 4-1 last May, saw revenues of €549.5m (£459.5m, at average exchange rate for the year ending 30 June 2014).
The club saw revenue growth of €30.6m, from increases of 8% and 9% in broadcast and commercial revenues respectively.
Other findings include:
Meanwhile, broadcast rights deals pushed the entire Premier League into the wider top 40 highest earning clubs.
"The Premier League's new broadcast deals have translated into big revenue increases across the English top flight," said Austin Houlihan, senior manager at Deloitte.
"In fact, every Premier League club reported record revenues in 2013-14."
DELOITTE FOOTBALL MONEY LEAGUE
•1. Real Madrid: 549.5m (518.9m)
•2. Man Utd: 518m (423.8m)
•3. Bayern Munich: 487.5m (431.2m)
•4. Barcelona:  484.6m (482.6m)
•5. Paris Saint Germain; 474.2m (398.8m)
•6. Manchester City: 414.4m (316.2m)
•7. Chelsea: 387.9m (303.4m)
•8. Arsenal: 359.3m (284.3)
•9. Liverpool: 305.9m (240.6m)
•10. Juventus: 279.4m (272.4m)
Source: Deloitte, revenues in euros for 2013-14 season.  Note: 2012-13 revenues in brackets
Mr Houlihan said that between them, the eight English clubs in the top 20 achieved total broadcast revenues of €1.1bn.
"The fact that all the clubs in the Premier League are in the top 40 is testament to the huge appeal of the league globally and also the equality of the distributions the clubs enjoy relative to their European counterparts," he added.
"Additionally, the Premier League is currently negotiating for the next cycle of media rights and further uplifts are anticipated."
Once again, the list was dominated by clubs from the so-called "Big Five" leagues; England's Premier League, Germany's Bundesliga, Italy's Serie A, Spain's La Liga and France's Ligue 1.
Real Madrid has matched its 10th triumph as champions of Europe by topping the Deloitte football rich list for a 10th straight year.