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Frank Schleck, the elder of the two Luxemburger brothers prominent in professional road cycling over recent years, was withdrawn by his team from the Tour de France on Tuesday night after an announcement that a urine test following Saturday's stage had revealed the presence of Xipamide, a banned diuretic.

Schleck, 32, and his brother Andy, 27, made history last year when they became the first siblings to appear on the final podium of the Tour in Paris. Andy was second and Frank third behind the winner, Cadel Evans. Andy had been awarded victory in the 2010 edition of the race after that year's winner, Alberto Contador, was suspended and had his titles stripped following a positive test for clenbuterol but has missed this year's event through injury.

The winner of two individual stages of the Tour, in 2006 and 2008, Frank Schleck experienced a difficult first two weeks of this year's race, in which his brother did not compete as a result of an accident in the Dauphine stage race last month. Frank was lying in 12th place, 9min 45sec behind the leader, Bradley Wiggins, before his removal.

Xipamide is a sulfonamide used for the treatment of oedema, fluid retention and hypertension. The governing body, the Union Cycliste Internationale, informed Schleck of the result of the test on the A sample during the afternoon of the Tour's second rest day and requested his removal from the race. He has four days to request a test on the B sample.

His Radioshack Nissan Trek team issued a statement confirming the rider's withdrawal. "Even though an abnormal A sample does not require these measures, Mr Schleck and the team believe it is the right thing to do, to ensure the Tour de France can go on in calm and that Frank Schleck can prepare his defence in accordance with the legal timing to do so.

"On the subject of Xipamide the team can declare the following: it is not a product that is present in any of the medicine that the team use and the reason for the presence of the Xipamide in the urine sample of Mr Schleck is unclear to the team. Therefore the team is not able to explain the adverse findings at this point. However, the team is fully determined to co-operate with the anti-doping authorities in order to resolve the matter."

The brothers are the sons and grandsons of racing cyclists. Their grandfather, Gustav Schleck, rode in the 1930s and their father, Johnny, rode in seven Tours as a domestique and was in the teams of the 1969 winner, Jan Janssen, and of Luis Ocaña, the winner in 1973. In recent years the Luxembourg flags carried by their supporters have become a familiar sight on the Tour.

Frank Schleck had never given a positive test before Saturday but during the 2008 Tour – in which the elder brother wore the yellow jersey for two days – the German newspaper Suddeutsche Zeitung claimed to have evidence of a payment of €7,000 three years earlier from Frank to the notorious doctor Eufemiano Fuentes, whose alleged involvement in doping was the subject of the Operación Puerto investigation in his native Spain. Fuentes denied any wrongdoing and said that his methods were perfectly legal.

Earlier in the race Johnny Schleck's car had been stopped and searched by French police but nothing was found.

Frank Schleck admitted making the payment to Fuentes, insisting it was simply for training advice. Temporarily suspended by his team, he was later cleared by the Luxembourg cycling authorities.

Radioshack Nissan Trek was created this year by a merger of a team set up by Lance Armstrong in 2010, during the second and last year of his comeback, and Leopard Trek, founded as a Luxembourg national project last year to provide a platform for the Schleck brothers. Their preparations for this Tour were accompanied by a public disagreement with Johan Bruyneel, formerly Armstrong's directeur sportif, who had taken over the same role in the new team.

This is the second drug incident to have emerged during the present Tour. The first, made public on the rest day following the opening week, resulted in the suspension of Rémy di Gregorio, a 26-year-old French rider with the Cofidis team, who was charged with "possession of banned substances or illegal devices". Di Gregorio insisted he has never doped.