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The companies bringing the case -- Dow Chemical Canada, Imperial Oil and Nova Chemicals, backed by Alberta and Saskatchewan provinces -- were expected to argue that Ottawa failed to provide scientific evidence to justify the regulations.
They will also try to convince the court that the federal government overstepped its jurisdiction in regulating waste management.
Environment Minister Steven Guilbeault in a Twitter message accused big multinationals of "trying to stop our ban on harmful single-use plastics."
"We're going to stick to the science and fight for a healthy environment where our shorelines, parks, and waterways are free of harmful plastic pollution," he vowed.
The phased-in ban on the import, manufacture and sale of plastic items such as store bags, straws, cutlery and beverage ring carriers was introduced last year as part of Ottawa's commitment to zero plastic waste by 2030.
It relied on a recent scientific assessment that found plastic pollutes waterways and oceans and can linger in the environment for hundreds of years, harming wildlife and people -- with microplastic fragments now found everywhere.
Canada produces more than four million tonnes of plastic waste every year, according to environmental group Oceana Canada.
Ottawa's ban has the widespread support of Canadians, and is in step with more than 50 other countries that are passing laws to fight plastic pollution, the group said in a statement.
"The continued production of single-use plastic is unsustainable and will prohibit Canada from meeting its commitment to zero plastic waste," Oceana campaigner Anthony Merante said.
"If the plastic industry wins this lawsuit, we will continue to see whales washing ashore, turtles drowning and more plastics in our food, water and blood," he warned.
After three days of hearings, a ruling on the matter is expected to be announced in the coming months.
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