A total of £251, 270 was collected at Leeds in 2015/16, down slightly on the £253,868 handed over during the previous year.
It contributed towards a total income of £5.46m from toilets at 12 of Network Rail’s stations, including Liverpool Lime Street, Manchester Piccadilly and Edinburgh Waverley.
Network Rail charges 40p for access to the toilets near the New Station Street entrance at Leeds, generating an average of £688 per day in the process.
The majority of its stations only charge 30p, although all toilets at Birmingham New Street are now free to use.
It means Leeds has the joint second highest charge alongside Glasgow Central, where facilities also include showers.
The figures show that only London’s Victoria Station charged more at 50p per visit.
Anthony Smith, chief executive of the independent watchdog Transport Focus, said: “Passengers dislike having to pay to use toilets at stations, particularly when they have forked out on a train ticket. It is important that money raised is spent on maintaining and improving facilities at stations.”
Revenue made in Leeds is used to cover the cleaning and maintenance of the paid-for toilets as well as the free facilities on platforms eight, 12 and 15.
A spokesperson from Network Rail said: “We do not profit from toilet charges. The small charge we make for using the toilets helps to maintain them and prevents misuse such as vandalism and other anti-social behaviour.
“Any money raised from the charges is reinvested into the railway and passenger facilities at our stations.”