The analysis by UKHospitality Scotland showed an average Scottish pub will be £15,000 worse off than its equivalent in England while a medium-sized hotel will be £30,000 worse off.
The trade body said larger businesses have been denied support worth up to £110,000 – the payment cap.
Despite calls from the Scottish hospitality sector for business rates support ahead of the Budget in December, finance secretary Shona Robinson failed to meet the calls and replicate the help for pubs in England with a 75% business rates discount, which Chancellor Jeremy Hunt announced in November.
UKHospitality Scotland cited information from The Fraser of Allander Institute that at least 10,000 hospitality firms were now operating without any financial assistance.
The latest figures from the trade body are set to be highlighted in an evidence session to the economy committee today (Wednesday 24 January).
UKHospitality Scotland executive director Leon Thompson said: “These figures clearly illustrate the real-life consequences of the Scottish Government’s decisions.
“In the current climate, it is almost impossible to fathom a local pub landlord or hotel manager being able to find themselves thousands of pounds to pay a bumper business rates bill in April.
“Many are struggling to keep the lights on as it is, in the face of extortionate rises in energy, food, drink and wages.
“It is an active choice of the Scottish Government not to support these critical venues and leave them significantly worse off than their English counterparts for the second year in a row.”
The trade body boss went on to highlight how the hospitality venues are pillars of their communities.
He added: “They are where we go to meet friends and family, celebrate an occasion or for some much-needed relaxation.
“There is still time for the Scottish Government to put right their widely criticised decision not to provide business rates support this year.
“As they finalise this year’s Budget, I would urge them to use the funds available to them and introduce a 75% business rates relief scheme.”