Barkley began his career with the Toledo Blue Stockings of the Northwestern League, and was a member of their championship team in. He was still with the team when they joined the American Association for the season, and was the everyday second baseman. He batted.306 that season and led the league in doubles with 39. Following the season, Toledo and the St. Louis Browns made an arrangement in the off-season for a trade involving several players, but the tradebroke down after the waiting period and only Barkley and one other player actually played with St. Louis. After a lawsuit it was estimated that Barkley had been valued for $800. Chris von der Ahe later claimed that Barkley's value was $1,000, but that may have been the asking price. In March 1886, Browns owner Chris von der Ahe offered Barkley for $1000 to the first team to pay the money. Billy Barnie was able to have Barkley sign an undated contract with the Baltimore Orioles and wired the asking price to Von der Ahe, but he had already secured a deal with owner of the Pittsburgh Alleghenys, Denny McKnight, and Sam was convinced to play for the Allegheny club instead. The American Association suspended and fined Barkley for signing with Pittsburgh this action. Barkley sued the Association, but they settled out of court with suspension being lifted although the fine stayed in place. Baltimore was offered and accepted Milt Scott as payment. That first season with Pittsburgh, the season, he hit.266 with 31 doubles, and he also stole 22 bases, while playing in 122 games. He stats declined significantly in, only playing in 89 games, hitting only.224. After the season was over, Pittsburgh sold him to the Kansas City Cowboys of the American Association. He was given good playing time in by the Cowboys, playing in 116 games, but his batting average slid further down, to.216, but the season was not uneventful. On June 13, he hit for the cycle, and he was given the managerial reins, which lasted 58 games and 21 wins.
Post-career
After his career in baseball ended, Barkley became a cigar maker. He died at the age of 53 in his hometown of Wheeling, and was buried in Peninsula Cemetery.