While the final tally has yet to be determined, it is expected that between $13,500 and $14,000 will have been raised for the Dunedin Hospital children's ward.
Great weather meant the walk was faster than expected, with the more than 370 people taking part completing it in about one and a-half hours.
A member of the Lions Club of Dunedin South which organised the trip, Graeme Elliot, said it had been thought it could take two hours as, although the route was reasonably flat, walking along railway tracks was not easy, particularly in the tunnels.
He described the day as "really fantastic", with everybody happy to have the opportunity to walk from Flat Stream to Deep Stream.
Walkers caught the Taieri Gorge train in the morning and were dropped off at Flat Stream.
The route took in four bridges/viaducts and two tunnels.
They then boarded the train again in the afternoon at Deep Stream and returned to Dunedin.
Mr Elliot said walking allowed people to see the superb stonework on the bridges and imagine what it must have been like for those building them in cold and horrible weather.
While a few people had bumps and grazes by the end of the walk, and one woman had fallen in a tunnel and put a tooth through her lip, St John Ambulance staff were on hand to deal with any concerns.
Tickets had been in great demand and it had been suggested that the club, which was helped in the venture by Peninsula Lions Club members, could organise another trip this year.
However, it was considered it was better to do it once a year and do it properly, Mr Elliot said.
The trip could not be carried out in summer because of the train schedules.
