NHS Fife is thought to be the first in Scotland to stipulate both partners must be non-smokers, as well as the female having particular body mass, before they will even be considered for the treatment.
A charity that helps infertile couples criticised the move to restrict the criteria, and a pro-smoking lobby group said it was discriminatory.
The change comes into effect from the start of October as a reduction in treatment cycle provision is also made in Fife.
Couples in the region trying to have a baby on the NHS through the process are to receive two instead of three cycles – a move that follows health boards in Dumfries and Galloway and in the Borders.
The Infertility Network charity said other boards have some restrictions on females smoking, but this is the first time it has encountered both partners being banned.
NHS Fife claimed the move, along with further funding of £100,000, would mean waiting times are reduced.
Gwenda Burns, of the Infertility Network, said the new criteria vastly narrowed couples' chances.
"There are health reasons involving smokers but, before, women have been offered help to stop smoking and they could go through the treatment," she said.
"Both partners being non-smokers is not in the guidelines."
Simon Clark, of Forest, which promotes smokers' rights, said: "It is discriminatory, and smokers have already paid into the welfare service through tobacco taxes, so in one way they have paid for the right to treatment."
The Infertility Network blamed the reduction on tightening health board budgets, and also said NHS Fife should have waited for a review of guidelines on provision of IVF due in the near future.
Ms Burns added: "This is bad news for patients. The decision to cut patient provision before the infertility review is finished is completely incomprehensible.
"To make this decision, without any consultation or warning, whilst we are so close to the end of this important review, is beyond belief. It is scandalous that health boards in Scotland continue to reduce provision and make changes to criteria knowing that new recommendations will soon be in place.
"This is the third health board to do so, with Dumfries and Galloway and Borders reducing provision to two cycles over the last year or so."
Dr Brian Montgomery, NHS Fife Medical Director, said: "NHS Fife has committed an extra £100,000 to fund infertility treatment over the next financial year.
"This will see 28 more cycles carried out than the previous financial year, helping to reduce waiting times for treatment.
"Treatment criteria have been revised to improve the success of the treatment and the outcomes for mothers and babies.
"Patients will receive up to two treatment cycles."
Dr Montgomery added: "Both partners must be non-smokers and the female body mass index should be less than 30kg/m2.
"These changes will come into effect for all new patients from October 2012, or those on the waiting list who are likely to be offered a cycle after April 1, 2013. Couples currently waiting for a third cycle will still receive this."