The Argentine rifled home from the spot to break another record by scoring for the 12th consecutive Liga game, cancelling out Ever Banega's opener for the resilient hosts
Ernesto Valverde had set out to refuse the Blaugrana their usual time on the ball and it was a tactic that looked to have worked superbly as Ever Banega put them ahead. However, Lionel Messi equalised from the spot to score in a record 12th consecutive Liga fixture.
Both teams opened the game with a lot of energy, but Barca looked more threatening as they were largely in control of possession. However, they were unable to fashion any chances to test Vicente Guaita as los Che defended in numbers and pressed quickly.
Valencia complemented their enthusiastic defence by playing on the break when they managed to wrest the ball away from their opponents, with Roberto Soldado keen to make an impact.
Midway through the first half the striker's bright run came to an end on the edge of the box due to a trip from Javier Mascherano. Tino Costa struck the free-kick but he failed to get it past the wall and his team's appeals for a Xavi handball were ignored.
Their resilience at the back continued to frustrate the Blaugrana and Valverde's men were rewarded in the 33rd minute. Soldado's cross was headed away by Gerard Pique, but only as far as Banega who brought the ball down and rifled a low shot past Victor Valdes at his near post.
Barca were handed a route back into the game as Joao Pereira clumsily cut down Pedro in the box in the 39th minute. Messi stepped up to take the penalty and fired an unstoppable effort into the top corner for his 34th Liga goal of the season.
The teams went into the break level but with just five second-half minutes played los Che nearly regained the lead. Andres Guardado's cross was met by Soldado but he could not guide his header inside of Valdes' post.
The game continued to go from end-to-end and five minutes later, Cesc Fabregas really should have put the Catalan side ahead after Messi's defence-splitting through-ball, though his shot only rippled the side netting.
Valencia were showing no signs of tiring as they continued to press the Barcelona players with great intensity, keeping the visitors out of their usual rhythm.
Sergio Canales replaced Banega in the 64th minute and it did not take long for the former Real Madrid man to make his presence known. From Guardado's cutback a fantastic, arrowed shot looked to be heading for the top corner until Valdes fingertips turned it wide of the post.
The keeper was once again in fine form in the 89th minute making a brilliant double save - the second of which came from Soldado - which stopped the hosts from stealing all three points at the death.
Nevertheless, Valencia secured a well-earned point that could also see Barcelona's advantage at the top of the table cut to nine points if Atletico Madrid defeat Betis on Sunday.