"I have accepted his resignation with effect from February 19," the Speaker informed the Lok Sabha.
Unprecedented pandemonium had broken out in the House last week when Rajagopal, who is opposed to the division of Andhra Pradesh, brought in a canister and sprayed pepper.
All political parties had demanded action against him and the government had said it would not oppose any decision taken by the Speaker.
The 50-year-old Vijayawada MP had yesterday sent his resignation to the Speaker hours after the controversial Telangana bill was passed by the Lok Sabha.
"I submit my resignation from Lok Sabha. I am quitting politics too. I'm hurt as Telugu-speaking people are divided now. It is a sad day. I lost interest in politics," Rajagopal told PTI yesterday.