The 29-year-old was substituted with 14 minutes remaining of Monday's 6-2 win over Iran after suffering a heavy tackle from Morteza Pouraliganji early in the second half and was pictured with strapping on his leg as he left the Khalifa International Stadium.
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Although manager Gareth Southgate insisted Kane was "fine" after the match, sources have confirmed that the England captain will undergo a scan on Wednesday.
It is unclear at this stage whether the scan is precautionary or a consequence of Kane feeling more discomfort. England held a light warm-down session at their Al Wakrah training base on Tuesday in which the players who started a day earlier against Iran did not feature.
The Tottenham striker has a history of ankle ligament injuries, suffering them during both his 2018 and 2019 Premier League campaigns.
Kane has scored 51 goals in 76 international appearances and needs two to match Wayne Rooney's England scoring record.
He won the Golden Boot as England reached the semifinals in Russia four years ago but did not score against Iran.
In other England news, James Maddison was missing once again as he recovers from an ankle problem while Callum Wilson, who replaced Kane as a substitute, skipped the outdoor session to concentrate on gym work.
Harry Maguire is also being monitored after being substituted due to illness.