Australian Nursing and Midwifery Federation members walked off the job at the Royal Hobart Hospital for 15 minutes on Wednesday afternoon.
The union, which rejected an offer from the state government to bring forward enterprise agreement talks, wants better pay and more clinical resources.
"It is time to resource the Royal Hobart Hospital and the Tasmanian Health Service safely," the federation's Tasmanian secretary Emily Shepherd said after the rally of several hundred people.
"We don't want to sit down and negotiate existing entitlements that need to be met.
"There is an obligation on the Tasmanian government to (ensure) safe staffing ... (and) safe workloads, and ensure our members aren't working an onerous amount of double shifts and overtime."
The island state's health system has been under increasing pressure amid a spike in COVID-19 cases.
There were 170 people with the virus in hospital on Wednesday, more than triple the figure at the beginning of July.
Strikes are are planned at the Launceston General Hospital and at facilities in the state's northwest in coming weeks.
"We're hoping the government will come to the table with concrete solutions we can put back to members and potentially call off our strike action," Ms Shepherd said.
"If not, our members have resolved at the Royal Hobart Hospital to escalate their strike action."
Andrew, a nurse for 12 years, said patients with serious mental health issues were waiting unprecedented times in the hospital's emergency department.
"We really care about patient safety and Tasmania having a good healthcare service. We're invested," he said.
He said there was an over-reliance on agency nurses.
"It's important to have a permanent fairly stable workforce that is paid appropriately."
Premier and Health Minister Jeremy Rockliff urged the union to sit down with the state government and negotiate in good faith.
"We've got increasing demand. At the moment there are a lot of pressures," he told reporters.
"I do not need strike action to hear the voice of nurses. I understand. I've read the comments, I've visited hospitals.
"Strike action will put further strain on our health system, which no one needs."
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