Tigers duo Sydney Stack and Callum Coleman-Jones were both injured in a fight early on Friday morning.
The AFL is investigating their COVID-19 protocol breaches, with Richmond facing the prospect of a $100,000 fine for a second misdemeanour.
It follows an incident in July where the wife of Tigers captain Trent Cotchin – Brooke Cotchin – broke the league’s strict biosecurity rules.
Stack and Coleman-Jones are facing the prospect of lengthy suspensions and Palaszczuk called for tough penalties.
It comes just two days after the Queensland premier delighted in a joint announcement with the AFL that the 2020 grand final would be held at the Gabba in Brisbane.
“AFL players caught breaking COVID rules should be sent home,” Premier Palaszczuk tweeted on Friday.
“Queensland won’t tolerate it. I know the AFL takes these issues seriously and will take appropriate action.”
A Queensland Police investigation is ongoing, with the AFL expected to hand down its own penalty at some stage on Friday.
In a club statement, Richmond said they immediately reported the players’ protocol breach to the AFL and are assisting the league with its investigation.
A second protocol breach could leave the Tigers’ hunt for a third premiership in four seasons in a precarious position.
Under AFL rules, third and subsequent breaches are punishable by the loss of premiership points, loss of draft picks or further financial sanctions.
A second breach carries a possible fine of up to $75,000 and the Tigers have another $25,000 hanging over them from Cotchin’s misdemeanour.
Stack, 20, has played nine of his 26 AFL games this season while Coleman-Jones, 21, made his one senior appearance last year.
The pair are currently listed as injured with respective hip and foot complaints.
They are not part of a separate quarantine hub where AFL officials and some players and family members are undertaking the required 14-day quarantine period.
Hundreds of AFL players and club staff are living under strict protocols in Queensland bubbles to complete the 2020 season.
The majority of them have been temporarily relocated from Victoria.