Act One

In Dr. Kolenaty's law offices, his clerk, Vitek, is filing papers and muses on the case of Gregor versus Prus, in inheritance case which has been unresolved since 1827. Baron Prus died without any acknowledged legitimate children and without a written will, but the Gregor family claims to be his descendants and rightful heirs, rather than Jaroslav Prus, the other claimant, and descendants of the "Mach Gregor," whom the Baron said he would make his heir. Albert Gregor comes in to ask about any progress. They are interrupted by Vitek's daughter Kristina's arrival. She hopes to become a singer and raves about the great Emilia Marty. Dr. Kolenaty himself enters, bringing Emilia Marty with him. She says that she is interested in the Gregor versus Prus case herself, and when Dr. Kolenaty starts to summarize it, shows as much command of the details as he possesses. She tells them that the missing heir is Ferdinand Gregor, the old Baron Prus' son by Ellian MacGregor, an opera singer. When they ask for any kind of proof, she says that the Baron's lost will, leaving the estate to Gregor, are mixed among the love letters now in Jaroslav's possessions, and Kolenaty is sent to fetch them, though he is dubious that this is the breakthrough she claims it is. Alone with her, Gregor tells her he loves her and asks for more information about his parents. She rejects him, but does ask him to give her any documents, written in Greek, that he might find among the family papers. Kolenaty returns, saying that Emilia was right, but Prus, who comes with him, asks for further proof. Emilia says she has this as well.

Act Two

At the opera house, staff gossip about Emilia. Janek, Jaroslav's son, and Kristina appear, and she flirts with him, but tells him that her career as a singer must come first. Prus enters, looking for Emilia, and Janek and Kristina are overwhelmed by Emilia's presence when she enters. She is irritable, however, and insists on paying Gregor for some jewelry he had given her and asks Janek and Kristina if they have "enjoyed heaven" together, though she says it's never worth it. Count Hauk-Sendorf enters and exclaims at the resemblance between Emilia and a Spanish singer, Eugenia Montez, who had once been his mistress. When she left him, he lost his mind. She seems oddly moved and sings to him in Spanish. She then sends everyone away and Prus questions her about her real name. He wonders why the initials keep "E.M." keep recurring: Ellian MacGregor, Eugenia Montez, Emilia Marty, and the name of the mother in the original documents about Prus' son, Elina Makropulos. He shows her another packet of papers, which he has not opened yet, and she asks to buy them. He leaves, and Gregor returns, again telling her he loves her. She begs him to steal the papers for her, if he truly loves her as he says he does, and he becomes violent and threatens to kill her. She shows him a scar on her neck from another man's murder attempt, and falls asleep as he continues his avowals of love and he leaves. Janek enters, and she now asks him to steal the papers. He is just about persuaded but Prus interrupts. He dismisses Janek and agrees to give her the papers, unopened, if she will sleep with him that night.

Act Three

In her hotel bedroom, Emilia and Prus are dressing. Prus gives Emilia the packet, though he says that she lay in his arms like a cold, dead thing, and she examines it; the contents are indeed what she was hoping for. One of Prus' servants comes to tell him that Janek has killed himself over his hopeless love for Emilia, but she is completely indifferent. Hauk-Sendorf comes to ask her to go to Spain with him, and she agrees, but when the rest of the cast comes in, with a doctor, the doctor leads him off. Kolenaty accuses her of forging the letters proving Gregor's claim; the handwriting is exactly the same as hers. She contemptuously tells him it was indeed Ellian MacGregor who wrote the letter and goes off into another room to finish dressing and have a drink, and they search her luggage. She returns, drunk, and tells Gregor that she is his great-great-great-great grandmother. She was born in Crete in the sixteenth century, and her father, Hieronymus Makropulos, was Emperor Rudolf's physician. He promised the Emperor an elixir that would prolong his life for 300 years, and Rudolf insisted that she drink it, to prove that it was not poisoned. After she drank it, she fell unconscious and her father was arrested. However, she awakened and since then, has had dozens of different identities. One of these was Ellian MacGregor, and she bore Prus a child, Ferdinand, Gregor's ancestor. Now she will age and die unless she drinks more of the elixir; she had lost the formula and has finally found it in the packet she obtained from Prus. She begins to wither physically, and her mind wandering, prays in Greek, and they begin to believe her story. Now that she has the recipe, and can prolong her life yet again, she has no wish to do so. Life has meaning only if it has an end. She gives the recipe to Kristina, who burns it, and Emilia dies.

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