"I'm done with Test cricket and won't be coming back for England's tour of Pakistan in December," said the all-rounder.
In the summer, the door was open for a return to the Test team for Moen, who retired from the game a year ago.
However, he told the Daily Mail he could not commit to the tour and the period in secure hotels it requires.
"Baz phoned me and I said, 'Sorry, I'm done,'" he said.
I don't see myself being stuck in hotels for another month and playing to the best of my ability.
Test cricket is tiring. Something has to give as I'm 35 years old. It wouldn't be right for me to reverse my decision and then struggle to give it my all, since I want to enjoy my cricket.
In 64 Tests for England, off-spinner Moeen has taken 195 wickets and scored 2,914 runs.
England will play two Tests in Pakistan on December 1st.
Chris Silverwood and Joe Root were England's coach and captain respectively when Moeen retired from Tests, but this summer has seen a sharp upturn in fortunes with six wins from seven.
During the South Africa series in August and September, Moeen was considered a replacement for a concussion or Covid-19 injury.
While watching the boys, I got the feeling this is how I've always wanted to play the game, but the more I thought about it and discussed it with those close to me, I knew I was done.
I'm closing the door on that side of my career. Being able to play 64 Tests for England has been a great experience.
England don't have a lot of options when it comes to spin bowling.
Parkinson was left out of the squad to play India and South Africa in the summer despite making his Test debut as a concussion substitute against New Zealand.
There are two left-armers who played for the England Lions against the South African team.
Will Jacks, who made his T20 international debut in Pakistan, is one of the candidates.
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