The 49-year-old faced Latrobe Valley Magistrates Court via video link from prison on Tuesday with her hair tied up, wearing a blue jumper and glasses.
She is accused of killing her former in-laws, Don and Gail Patterson, both 70, and Gail's sister Heather Wilkinson, 66.
All three died in hospital days after consuming an allegedly deadly mushroom meal at Patterson's Leongatha home, in South Gippsland, on July 29, 2023.
She is also accused of the attempted murder of her ex-husband Simon at the lunch and on three occasions dating back to 2021, and the attempted murder of Ms Wilkinson's husband Ian, 68.
Patterson pleaded not guilty to all eight of her charges on Tuesday, including three counts of murder and five counts of attempted murder.
She was remanded in custody to face trial at the Supreme Court in Melbourne after she elected not to have a committal, fast-tracking the court process.
Her case was next listed for a directions hearing at the Supreme Court on May 23.
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