Riding from Kalgoorlie to Perth, teams will spend five days from September 27 to October 1 following the Golden Pipeline.
The trail has been designed so it is suitable for serious or casual recreational mountain bikers, novices and families.
The challenge will raise funds for Joondalup-based charity Youth Futures.
Each rider will have a minimum fundraising target of $1000 and all money raised will go towards accommodation, education and support services for young people who are dealing with issues such as mental health, homelessness and abuse.
The challenge will also include an e-bike category, where participants design and build a 250W battery powered bike that will travel each stage of the challenge, with the longest distance being 180km. For details, go to www.pipelinechallenge.com.au.