Speaking at the Scottish Grocers Federation (SGF) conference last week, SGF chief executive John Drummond said the convenience sector would witness its "biggest change in a generation" when the Licensing (Scotland) 2005 Act comes into effect in September.
In addition to increased application fees, retailers will have to complete a layout and operating plan and ensure staff are appropriately trained before being granted a licence.
In a poll of SGF members, estimates of how much complying with the new regulations would cost varied from £1,500 to £5,700 a store.
"When the Scottish government gave assurances that retailers operating responsible businesses had nothing to fear from the new legislation this obviously did not apply to costs," Drummond said.
Under the regulations, anyone who authorises retail sales of alcohol in off-licence premises in Scotland must hold a personal licence certificate.
Course provider CPL Training said "hundreds" of retailers were taking the personal licence holders course - which costs about £125 per person - but warned others not to delay in getting the qualification.
"We are advising people to take the course as soon as possible to avoid the inevitable mad scramble in the summer," said trainer Aileen McDonald.
Nassar Saddiq, who owns two stores in Glasgow, took the course recently with eight of his staff. "It has definitely improved our judgement and we are much more vigilant now," he said.