Chelsea were trailing 1-0 to a Rodrigo goal when they were awarded a penalty thanks to VAR with the chance to level matters against Valencia.
Substitute Barkley immediately picked up the ball and strode towards the spot, and continued to wave away Jorginho, Willian and Tammy Abraham.
Barkley then missed the ensuing spot-kick, rattling the crossbar but after the game Lampard defended his player.
Chelsea's English midfielder Ross Barkley (L) hits a penalty over the bar during the UEFA Champion's League Group H football match between Chelsea and Valencia at Stamford Bridge in London on September 17, 2019.
Image credit: Getty Images
"Ross is a penalty taker and is when he starts games. He took it and missed it." Lampard told BT Sport.
It is a great story to say there is contention between players. If he scores it is not a story. But he took it and missed. That is it.
“The disappointment to all of us is that we didn't get chance to win the game.
“There is no issue in the dressing room."
Lampard doubled down on that line during his press conference.
"Ross is the penalty taker... Jorginho and Willian are the penalty takers on the pitch. When Ross comes on, he's the penalty taker."
Another Chelsea midfielder Mason Mount had to be substituted during the first-half after a dangerous challenge from Francis Coquelin.
Lampard admitted that losing the in-form youngster impacted Chelsea’s play.
"It did affect the game and we know how well he is playing.” Lampard said.
“He looked sharp. It is an ankle injury and hopefully not too bad. It needs to be assessed over the next few days."