Alessandro Petacchi will "definitely not be racing" in the Tour de France, a spokesman from his team Milram said.
Petacchi has suffered from asthma for many years
The news comes after the Italian Olympic Committee charged him with doping at this year's Giro d'Italia and he will face a hearing on 24 July.
All 189 riders have been passed fit to start the Tour on Saturday after blood tests were carried out on Thursday.
"We have carried out 189 blood tests. There are no unfit cases," said an International Cycling Union spokesman.
Petacchi, who protests his innocence, faces a one-year ban after testing "non-negative" for salbutamol, which he is allowed to take for his asthma.
"Non-negative" means a substance is found which may be legal if the rider has a medical certificate.
Nevertheless, the Olympic Committee has asked the Italian Cycling Federation to ban him.
The International Cycling Union said the 33-year-old Petacchi, who is currently suspended by his Milram team, tested for salbutamol at Pinerolo on 23 May, the day he claimed the third of his five stage wins in this year's Giro.
"What can I do to demonstrate my good faith? I do not want to miss the Tour," Petacchi, who has 21 ProTour victories, told Italian newspaper Gazzetta dello Sport when the story first broke.
"Why would I have done something different from other times? I have the responsibility of the team on my shoulders, with the sponsors that are always speaking against doping."
Team Milram spokesman Andrea Agostini said Petacchi was 320 nanograms per millilitre above the authorized level of 1,000 nanograms per millilitre.
"This is not a significant amount and historically, as long as there is a satisfactory reason, cases of this nature are archived," he said.
The urine sample taken at Pinerolo was the only one of the five doping tests Petacchi gave that showed an excessive amount of salbutamol.
"I may have done one spray more but I have done nothing illegal," said the cyclist.
Italian cyclist Leonardo Piepoli has been suspended by his team Saunier Duval after testing positive for the banned substance salbutamol during the Tour of Italy.
The International Cycling Union (ICU) informed Saunier Duval on Thursday that the climbing specialist's test had produced higher-than-permitted levels of the drug which is used to treat asthma.
Last month Piepoli revealed that he did have medical authorisation to use the drug.
"I take salbutamol to relieve my allergies," argued the Italian rider who claimed the 'King of the Mountains' crown at this years Giro d'Italia.