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Arsenic and Old Lace is based on a play by the same name, written in 1939 by American playwright Joseph Kesselring (1902-1967). The script was adapted for the movie by American screenwriter Julius J. Epstein. A remake, also titled Arsenic and Old Lace (1969) was released in 1969.

Arsenic and Old Lace is a turn on "lavender and old lace," a term often used to describe the Victorian era when the homes of genteel older ladies were over-decorated with touches of lace doilies, lace curtains, and the smell of lavender dried and stitched into little bags of calico or gauze and hung up in wardrobes or put into dresser drawers. Also during this period, lavender-water was considered a suitable scent for elderly women, rather than a more seductive perfume, and shades of mauve or lavender were considered suitable to wear, enhancing a pale complexion and white or grey hair. Lavender and Old Lace was used in the title of a 1902 mystery and romance novel by American author Myrtle Reed. The movie Ladies in Lavender (2004) picks up this theme. In the case of Arsenic and Old Lace, the term "lavender" has been replaced with "arsenic" for obvious reasons... that's what these genteel elderly ladies use to poison lonely elderly men.

Those who have seen both the movie and the play say that there are several differences, the biggest one being that the play is set entirely in the Brewsters' living room, so there are no visits to the judge or the doctor. Also, the play is set in early September whereas the film is set on Halloween. The idea of Mortimer Brewster (Cary Grant) writing anti-marriage books was wholly a film invention; the play had him as a drama critic. Mortimer and Elaine are merely engaged in the play and have not just gotten married (and it's implied they've engaged in a bit of premarital fun, which would have been against Production Codes in the 1940s). Also against the Codes was the line from the play where Mortimer shouts triumphantly, "I'm not a Brewster! I'm a bastard!!!!". That line was changed to "I'm the son of a sea cook" for the movie. In the original Broadway play, (Boris Karloff) played Jonathan Brewster, but he wasn't available for the film, so Raymond Massey took over the role. Consequently, when the policeman tells Jonathan that he looked like Boris Karloff, it was a lot more meaningful in the play because he actually WAS Boris Karloff. Finally, the play has an entirely different ending where Mr. Witherspoon (Edward Everett Horton) from the sanitarium ends up being the aunts' final victim by sipping some of the wine as the curtain falls.

Karloff was playing the role of Jonathan Brewster in the Broadway play of Arsenic and Old Lace when filming of the movie was taking place in 1941 and was unable to be released for the filming. Consequently, Raymond Massey played the role in the film.

It is the third movement from Mozart's 11th Piano Sonata in A major. It's sometimes referred to as Rondo à la Turca or Turkish March.

Dr Einstein (Peter Lorre) is trying to convince Mortimer to leave so that he doesn't get hurt. Einstein slams the door and mutters "Mein Gott, was hab ich verbrochen?!", a German phrase used when one's situation is exasperating. It literally translates as "My God, what crime have I committed?" Figuratively, it means something like "My God, what have I done to deserve this?"

According to the Diagnostic and Statistical Manual of Mental Diseases, Teddy Brewster (John Alexander) has just that-a Delusional Disorder (297.1), more specifically called "Grandiose Delusional Disorder" in which a person believes themselves to be some prominent individual. It isn't all that common, but because it is quite entertaining to have a person imagine that they are the President or Napoleon or Jesus, the condition appears in movies and stories (and jokes) fairly often.

Jonathan Brewster is identified by a senior officer and taken away. Dr Einstein walks away under the noses of the police. Teddy is committed to the Happydale Sanitarium but Aunts Abby (Josephine Hull) and Martha (Jean Adair) don't want him to be alone, so they also commit themselves with Mortimer signing the commitment papers as the nearest relative. The aunts tell him he shouldn't really sign the papers because he wasn't their real nephew. He was the son of their cook, and his father was a sea cook (in other words, he isn't related to Teddy, Jonathan, and the aunts, who are obviously insane). In the final scene, Mortimer goes off with Elaine (Priscilla Lane), and the taxi driver never gets paid.

Viewers who enjoyed Arsenic and Old Lace say that His Girl Friday (1940), The Awful Truth (1937), and Bringing Up Baby (1938) come close in zaniness. In His Girl Friday, Cary Grant plays a newspaper editor trying to keep his ex-wife from remarrying. Similarly, in The Awful Truth, Grant plays a newly-divorced man trying to keep his ex-wife from remarrying while she is doing the same to him. In Bringing up Baby, Grant plays a paleontologist involved in a caper that includes a Brontosaurus bone, two leopards, and Katharine Hepburn.