Totti gave Roma a first-half lead against Sampdoria at the Olympic stadium only for the visitors to level through Gianni Munari after the interval to cement third place in the table.
The 35-year-old, who over the years has been used variously as a striker and a playmaker, joins Jose Altafini and Giuseppe Meazza in third place on 216 goals.
He is now only nine goals short of Sweden's Gunnar Nordahl, the former AC Milan and Roma forward who scored 225 goals between 1949 and 1958.
Silvio Piola, who played for Pro Vercelli, Lazio, Juventus and Novara holds the all-time record of 290.
All-time leading scorers in Serie A
1. Silvio Piola (Pro Vercelli, Lazio, Juventus, Novara) 290
2. Gunnar Nordahl (AC Milan, AS Roma) 225
3. Giuseppe Meazza (Inter, AC Milan, Juventus) 216
. Jose Altafini (AC Milan, Napoli, Juventus) 216
. Francesco Totti (AS Roma) 216