After graduating high school, Sam joined his father on tours playing saxophone and guitar. In Harare, he had his own jazz-influenced band called Ay Band with whom he recorded his debut album, Rume Rimwe, in 2008. He also had recorded two solo albums. The elder Mtukudzi introduced Sam as "the future" to appreciative crowds at a British nightclub in late 2009.
In the early hours of Monday, March 15, 2010, Sam and his sound engineer, Owen Chimhare, died in a car crash while traveling back to Norton near Harare. Their Tata pickup truck, driven by Chimhare, struck a bridge just before the Kuwadzana Extension off-ramp along the Harare-Norton road. The vehicle veered off the road, rammed into the right side of the bridge's guard rails, and ploughed into the river bank below. Inspector Tigere Chigome, a national Traffic Policespokesman, explained in the news that the accident happened at 1:20 AM. Both Mtukudzi and Chimhare died of head injuries at the scene.
Sam Mutudzi was an ambassador in the Dance4Life initiative whose national concept owner are Students Partnership Worldwide in Zimbabwe (SPW Zimbabwe). The initiative seeks to fight HIV and taboos surrounding it using the medium of music and dance. It is being run in several countries such as South Africa, The Netherlands, Kenya, Zambia, Tanzania, UK, US, Russia amongst others. Mtukudzi was identified as an ambassador by Luckmore jalisi from SPW Zimbabwe. He saw him as the appropriate person for the job due to his youthfulness as well as focus maturity in music.