The 'Super Bass' rapper surpassed Eminem on the micro-blogging site on Monday afternoon (January 9), with a total of 8.27 million followers.
This was compared to Eminem's 8.25 million followers, leaving him trailing closely behind in second place.
Minaj is currently preparing to release her second studio album Pink Friday: Roman Reloaded in the UK on February 13 and the US on February 14.
The star plans to unveil the music video for the LP's lead single 'Stupid Hoe' next week. She told her Twitter followers: "Looking @ the Stupid Hoe 1st edit....omfg. Ur gna spazzzz (sic)"
> Nicki Minaj: New album Roman Reloaded will snatch wigs and bald spots
Listen to Nicki Minaj's 'Stupid Hoe' below (Warning: strong language):