The tie at Madrid's Santiago Bernabeu will be the third of four matches between the arch rivals, clashes which are known as "El Clasico", in 18 days across three competitions.
Real recently defeated Spanish first division leaders Barca 1-0 in the Copa del Rey final on April 20 and it is comments made by Mourinho in the wake of that match which aggravated Guardiola.
"Since he has chosen to to talk about me, I'll do the same," former club captain Guardiola said on Barcelona's official web site.
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"[Wednesday] we'll face each other on the pitch. He's won his games off the pitch and I'll give him his personal Champions [League] for that -- just take it home and enjoy it."
Portuguese boss Mourinho used his press conference ahead of the match to accuse Guardiola, 40, of criticizing referees when they make correct decisions.
The former Inter Milan coach's comments related to the Copa del Rey final, in which Barca had a goal from Spain forward Pedro correctly disallowed for offside.
"There have been two groups of managers up until now," the one-time Chelsea coach Mourinho told Real's official web site.
"A quiet one who doesn't talk about referees and a louder one, such as myself, that criticizes the referees when they make significant mistakes.
"We have entered a new era following the statements made by Guardiola and there is now a third group. He is the only one in it as he criticizes the right calls made by referees. I had never seen anything like it before."
According to British newspaper The Guardian, Guardiola responded in strong fashion and said: "In this room [Real's press room], he is the chief, the f*****g man. In here he is the f*****g man and I can't compete with him.
"If Barcelona want someone who competes with that, then they should look for another manager. But we, as a person and an institution, don't do that."
Guardiola, who was a Barca player when Mourinho was part of the club's coaching staff between 1996 and 2000, was unhappy with his opponent alleging he had targeted match officials.
"I simply congratulated Madrid after the [Copa del Rey] final. If you think that his allegation that I always complain about the referees is true after you've all heard me over the last three years, well there's nothing I can do.
"Off the pitch, against his media supporters, there's nothing we can do to fight that. I've seen it before with Mourinho, he did it at Chelsea and now he's doing it here."
The Catalan club, who are three-time European champions, will be without Spain midfielder Andres Iniesta for the first match of the last-four tie.
The 26-year-old, who scored the winner for his country in the 2010 World Cup final against Netherlands, is out with an injury to his right calf, but club captain Carles Puyol should be fit for the match.
Real have won Europe's premier club competition on nine occasions, but will be without Sami Khedira for Wednesday's clash after the Germany international suffered a leg injury.
Portugal defender Ricardo Carvalho will also miss the match through suspension.
The return leg is in Barcelona on May 3, with the winner advancing to face Manchester United or Schalke in the final at Wembley Stadium, London on May 28.
Source: http://edition.cnn.com/2011/SPORT/football/04/27/football.mourinho.guardiola.clasico/index.html