The milestone journey was made at 4pm on Wednesday afternoon on bus route 322, running from Tesco in Brent Park to Paddington Station.
The fare allows passengers to take a £1.50 bus or tram journey and then change onto another bus or tram for free within one hour of touching in at the start of their journey. Anyone who uses pay as you go with a contactless bank payment or Oyster card has automatic access to it.
Around 200,000 journey also involved the London tram network in south London; 80,000 made the switch from a bus to a tram, with 65,000 changing from a tram to a bus.
Mayor of London Sadiq Khan said:
"I'm delighted that the Hopper fare is making such a difference to the lives of Londoners. We must ensure that everyone can afford to travel around our city, particularly people who rely on public transport to get to work or to study. Alongside our commitment to freeze all TfL fares, our Hopper fare is a crucial part of our plans to make it more affordable to live and work in London."
The Hopper fare was one of Sadiq Khan's manifesto promises, though when it launched last month it was given a slight rejig.
He had pledged a new one hour bus ticket allowing unlimited transfers on buses for that sixty minute period.
Those travelling on trains or tubes between two bus or tram services miss out. TfL said it hopes to roll-out unlimited transfers from next year, when the service will be expanded to include Tube and train journeys.
The most popular Hopper routes so far:
The 279 from Waltham Cross bus station to Manor House Underground station
The 53 from Orchard Road in Plumstead to Horse Guards Parade in Westminster
The 207 from the Hayes By-Pass near Southall to White City Bus Station in Shepherds Bush
The most popular bus stop to hop at is Brent Cross Shopping Centre, where more than 20,000 hops have taken place