"Hello to everyone from atop this toilet bowl," Matthews said before he launched into his performance.
Kimmel said that the bathroom offers the best acoustics when performing at home.
Matthews said that he always wanted to cover "American Tune" and was given enough time to study and learn it due to COVID-19.
Matthews also discussed how his family is doing at home during the pandemic.
"Being together without the possibility of going out and getting in contact with other friends, other than on, you know, screens, has tested our patience," he said.
"But we all come back thankfully, nobody has run out of the house screaming with their heads on fire," Matthews continued.
Kimmel has been producing this new version of Jimmy Kimmel Live from his home due to the COVID-19 pandemic. The late night host and Matthews promoted charity World Central Kitchen which uses food to empower communities.
Matthews has co-written a recently released children's book titled If We Were Giants which he says was inspired by New Orleans.