The superhero film, starring Andrew Garfield and Emma Stone, was a runaway hit with $65 million, nearly double that of runner-up Ted, which slipped from the top spot with a still-respectable $32.6 million.
Remaining in third place with $20.3 million was Disney Pixar's animated feature Brave, while Oliver Stone's drug war drama Savages entered the chart at number four with $16.2 million.
Channing Tatum stripper film Magic Mike fell three places to number five with $15.6 million, while Tyler Perry's Madea's Witness Protection came in just behind at number six, dropping three spots and bringing in $10.2 million.
Slightly under-performing during its opening weekend was Katy Perry: Part of Me, which entered the chart in eighth position and grossed only $7.1 million, coming in just ahead of number seven's Madagascar 3: Europe's Most Wanted.
Rounding out the Top 10 were Wes Anderson's Moonrise Kingdom, which fell two spots to number nine, and To Rome with Love, which made an impressive leap from 17 to ten and brought in $3.5 million.
The US box office top ten in full (studio estimates):
1. (-) The Amazing Spider-Man - $65,000,000
2. (1) Ted - $32, 593, 000
3. (3) Brave - $20,162,000
4. (1) Savages - $16,162,000
5. (2) Magic Mike - $15,610,000
6. (4) Tyler Perry's Madea's Witness Protection - $10,200,000
7. (5) Madagascar 3: Europe's Most Wanted - $7, 700,000
8. (-) Katy Perry: Part of Me - $7,150,000
9. (7) Moonrise Kingdom - $4,642,000
10. (17) To Rome with Love - $3,502,000