Charlene Doofenshmirtz is the ex-wife of Dr. Heinz Doofenshmirtz and shares custody of their daughter, Vanessa. She didn't divorce Heinz because he was evil; in her words, they just didn't get along very well and wanted different things. Whenever Vanessa tries to tell her that Heinz is actually evil, Charlene is adamant that no one can be evil. In contrast to Heinz's agitation with her, she is actually very civil in his presence and talks of and to him as if they were still on relatively good terms ("I Scream, You Scream", "Hail Doofania!")
Charlene has short dark brown hair and a slightly yellow skin tone. Her body-shape is similar to Linda's. She wears a green polo shirt with dark green buttons, black shoes, and blue jeans. She also has a yellow pearl necklace, a pair of yellow earrings and purple glasses.
She is a bit sarcastic with her ex-husband, and despite talking with him in good terms, she tends to make sarcastic comments about Heinz's ineptitude ("This Is Your Backstory", "Hail Doofania!"). She is also very innocent in subjects such as Heinz's schemes and -inators ("I Scream, You Scream") and never believes the schemes that her ex-husband has made, nor does she believe Phineas and Ferb's amazing inventions ("Finding Mary McGuffin").
However, she can be caring towards her daughter Vanessa, even personally encouraging her to donate to charity ("Finding Mary McGuffin").
- She and Linda Flynn-Fletcher are in the same cooking class and sometimes carpool. ("I Scream, You Scream", "It's a Mud, Mud, Mud, Mud World").
- She doesn't ever appear to take her glasses off for any reason, not even when she's getting treatments at a spa, including facial masks. ("Hail Doofania!").
- Despite her sarcasm towards Heinz's ineptitude, Charlene was briefly surprised that Heinz built in an -inator to help him with his poor kicking skills as he managed to pull a fantastic kick during the Doofenshmirtz family kickball game, much to everyone's surprise. ("Thaddeus and Thor") She also briefly complimented Heinz for using bee pheromones to make himself look attractive, to which he thanks her for. ("Bee Day") She is even perfectly aware that Heinz has built a giant robot named Norm to serve as his assistant, even complimenting Norm for his tuxedo design. ("This Is Your Backstory")
- She appears to have the same personality as Linda, though perhaps a little more sarcastic. Linda also at one point dated Doofenshmirtz before he met Charlene. ("What Do It Do?").
- Charlene shares another aspect with Linda: a daughter that wants to "bust" another person. In this case, it is Vanessa who wants to "bust" her father ("I Scream, You Scream", "Hail Doofania!").
- Though she and Heinz are divorced, she decided to keep the last name of "Doofenshmirtz" and attends Doofenshmirtz family reunions ("Thaddeus and Thor").
- She donates goods to charity and encourages her daughter to do so. Charlene also appears to be dating the man who drives the delivery van, Sam ("Finding Mary McGuffin").
- She is revealed to be rich as she pays Heinz a rather hefty alimony, which he uses to help finance his schemes and at one point pay for his building. ("Finding Mary McGuffin", "Run, Candace, Run", "Lotsa Latkes").
- She is possibly the same age as Linda, as their daughters are in the same age group.
- Heinz Doofenshmirtz said that one of the things that made them split apart was his desire to be evil (How to Conquer the Tri-State Area). However, he finally comes to the fact that he isn't as evil as he wants to be. (Last Day of Summer)
- She drives a car that strongly resembles a 1973 Chevrolet Camaro, which is erroneously identified by the Evil Jingle as a sports sedan. ("It's a Mud, Mud, Mud, Mud World", "Brain Drain")
- She had knee surgery. ("Candace Disconnected")
- She broke up with Doofenshmirtz because of a sockpuppet named Mr. Tomato. ("Bee Day")
- She is a Pisces, so she was born between February 19 and March 20. ("Love at First Byte")
- It was revealed that when Heinz and Charlene went to Community College together, a man named Lang (who is currently the principal of Danville High) had a crush on Charlene and developed a grudge against Heinz for winning her heart. Even after being told that Heinz and Charlene have been divorced, Lang stubbornly refuses to abandon his grudge. Whether or not Charlene returned Lang's feelings is never made clear. ("Doof 101")
- "I Scream, You Scream" (First appearance)
- "It's a Mud, Mud, Mud, Mud World" (Non-speaking cameo)
- "Hail Doofania!"
- "Thaddeus and Thor" (Speaking cameo)
- "Finding Mary McGuffin"
- "Brain Drain"
- "Rollercoaster: The Musical!" (Cameo)
- "Run, Candace, Run" (Pictured only)
- "Phineas' Birthday Clip-O-Rama!" (Pictured only)
- "Lotsa Latkes" (Pictured only)
- "This Is Your Backstory"
- "Bee Day"
- "Steampunx" (Cameo in song)
- "It's No Picnic" (Pictured only)
- "The Magnificent Few"
- "Run, Candace, Run"
- "Candace Disconnected"
- Phineas and Ferb the Movie: Across the 2nd Dimension
- "Night of the Living Pharmacists"
- "Doof 101"
- "Last Day of Summer"