More than £120,000 was raised during the Vélo Birmingham Business 100 challenge.
The 100-mile cycling sportive saw 15,000 cyclists take to the roads of Birmingham and the West Midlands last month and part of the event saw companies take part in a special business challenge.
More than £123,000 was raised for local and national good causes by businesses from across the Midlands, adding to the £2 million expected to be collected for charity by the event's individual participants.
The sportive's Business 100 participants began the day in an exclusive starting wave led by Vélo Birmingham ambassador and former England rugby captain Martin Johnson.
It was supported by the Greater Birmingham Chambers of Commerce, Birmingham Post and law firm DLA Piper.
With a total of £31,126 raised, the Queen Elizabeth Hospital Birmingham Eye Cycle Team won the prize of an all-expenses trip for four riders to take part in the 2018 Cape Argos, a annual cycle event held in Cape Town, South Africa.
Surgeons, patients and their families joined forces to raise money for sight-saving equipment as part of the hospital's Eye Appeal.
Tim Matthews, consultant at Queen Elizabeth Hospital, said: "It was a pleasure to cycle Vélo Birmingham alongside my colleagues.
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"We all enjoyed the significant challenge that cycling 100 miles posed to us and it was wonderful to raise so much money.
"We are all incredibly excited and honoured to have won the Business 100 prize and we look forward to sending a team over to South Africa next year."
Birmingham-based workplace management specialist Donseed raised £22,615.25 for their charities of the year and the team was joined by Mr Johnson as their honorary fifth member for the sportive.
HomeServe, including chief executive Martin Bennett, rode in aid of The Royal British Legion and its Pop-in Centre in Birmingham, raising £20,689.