Chitalu Chilufya, 48, was held in June following accusations he used ill-gotten gains to acquire property.
A court cleared him of the charges in August due to lack of evidence and he resumed his duties.
But President Edgar Lungu unexpectedly fired the minister on Sunday without given any reason.
“President Lungu has terminated the appointment of Chitalu Chilufya as health minister with immediate effect,” presidential spokesman Isaac Chipampe said in statement.
Chilufya’s sacking comes days after parliament quizzed officials over irregularities cited in an audit of financial statements released last year.
The reports allege the health ministry misused $17 million of public money meant to fund rubber gloves, condoms and medical kits.
Several opposition party leaders and civil society groups had since been calling on the president to remove him from office.
Chilufya has not yet responded to the accusations.
The minister is considered a potential candidate for presidential elections in August this year if Lungu is barred from running.
The president insists he will stand for a third term despite repeatedly failing to amend the southern African country’s constitution in order to do.