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Here is a list of the first episode appearances of characters who have appeared in the show at least two times.

Anchovies : Help Wanted
Barnacleboy : Mermaidman & Barnacleboy
Bubble Bass : Pickles
Dirty Bubble, The : Mermaidman & Barnacleboy II
Doctor Fish : I Was a Teenage Gary
Doctor Fish (Season 4) : The Lost Mattress
Don the Whale : Ripped Pants
Flats the Flounder : Sandy's Rocket
Flying Dutchman, The : Squidward, the Unfriendly Ghost
Fred : Reef Blower
Gary : Help Wanted
Goo Lagoon Event Announcer : Ripped Pants
Jellyfish : Help Wanted
Karen the Computer : Plankton!
Larry the Lobster : Ripped Pants
Lazy Mary : Chocolate With Nuts
Lou the Vendor Fish : Ripped Pants
Mama Krabs : Sailor Mouth
Manray : Mermaidman & Barnacleboy III
Mermaidman : Mermaidman & Barnacleboy
Mr. Doodles : Sandy's Rocket
Mr. Krabs : Help Wanted
Mr. SquarePants : Home Sweet Pineapple
Mrs. Puff : Boating School
Mrs. SquarePants : Home Sweet Pineapple
Patchy the Pirate : Christmas Who?
Patrick Star : Help Wanted
Pearl : Squeaky Boots
Plankton : Plankton!
Potty the Parrot : Christmas Who?
Sandy Cheeks : Tea at the Treedome
Scooter : Ripped Pants
SpongeBob SquarePants : Help Wanted
Squidward Tentacles : Help Wanted
Squilliam Fancyson : Band Geeks
Tom : Ripped Pants