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Sir Mark condemned the claims saying: "It's complete nonsense.
"We operate independently under the law without fear or favour.
"They are putting them at risk by suggesting that any of those officers are going out with, as with any intent, other than to operate without fear or favour in protecting communities."
Two-tier policing is used to describe the impression that some protests and demonstrations are dealt with more harshly than others.
Read more: What is two-tier policing and why does it exist?
The prime minister and the government have faced criticism from some on the right, including Nigel Farage, as well as X owner and billionaire Elon Musk, that police have handled the past week's disorder more harshly than if it had involved members of ethnic minorities.
Sir Mark said commentators from both ends of the political spectrum "like to throw accusations of bias at the police because we stand in the middle".
"If you've got crazy views over there, you don't like it. And if you've got crazy views from over there, you don't like it," he said.
"The serious voices who echo those are of more concern to me because the risk is they legitimise it, and they legitimise the violence that the officers I'm sending on mutual aid today will face on the streets."
Read more:
Three men jailed in first Crown Court sentences after riots
Musk v Starmer: What have they accused each other of in riots row?
More than 500 new prison places for rioters
Sir Mark refused to name any individuals as he said he did not want to "give them credit".
He added: "If you're a keyboard warrior, you're not safe from the law if you incite violence."
More than 6,000 extra police are being deployed across the country ahead of dozens of protests planned for this evening, including outside immigration help centres.
Police sources have told Sky News they're aware of more than 100 events and around 30 counter protests planned for tonight. Out of the 43 police forces in England and Wales, 41 are expecting gatherings.
More than 400 people have been arrested and about 100 charged in connection with the riots.
They started following the killings of three girls in Southport last month - and after false information spread online that the stabbings suspect was an asylum seeker who arrived on a small boat last year.
More defendants were due to appear in court today.
Three men jailed after disturbances which took place last week is evidence of "the swift action" the government is taking, said Sir Keir Starmer.
The prime minister wrote on X: "This is the swift action we're taking.
"If you provoke violent disorder on our streets or online, you will face the full force of the law."
Sir Mark said "protecting communities is at the core of policing" and forces have changed tactics in the last couple of days to relocate police officers at "key strategic locations" across the UK "to give us more agility to respond to wherever the protests are most intense".
"So that we can respond with agility and show overwhelming force, so that the criminals and thugs on the streets don't succeed."
The police chief echoed the director of public prosecutions' comments that terrorism charges should be considered.
"We'll throw the full force of the law at offenders, whether that's charging people with assaults, violent disorder, riot - and if terrorism offences are appropriate," he added.