The 49-year-old was confronted at Latrobe Valley Magistrates Court via video link from prison on Tuesday, with her hair tied back, 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 fatal mushroom meal at Patterson's home in Leongatha, South Gippsland, on July 29 last year.
She is also charged with the attempted murder of her ex-husband Simon over lunch and on three occasions dating back to 2021, and the attempted murder of Ms Wilkinson's husband Ian, 68.
Patterson pleaded not guilty Tuesday to all eight charges, including three murders and five attempted murders.
She was taken into custody to stand trial in the High Court in Melbourne, after choosing no bond, speeding up the judicial process.
Her case was then listed for a hearing at the Supreme Court on May 23.