The duo, who are taking part in the marathon event in aid of Comic Relief, eclipsed the previous time of 37 hours on-air - set by Simon Mayo in 1999 - at 7.30pm on Thursday.
They are now aiming to break the Guinness World Record, which currently stands at 50 hours. If they are successful, they will reach their target on Friday at 8.30am, but will attempt to stay on until the end of their regular breakfast show at 10am.
The pair have already raised more than £1 million for the Red Nose Day charity. Commenting on the rising total, Moyles declared: "That's amazing, just to watch us idiots and listen to us idiots for two days."
A number of celebrities, including Ricky Gervais, Davina McCall, Lily Allen and Jimmy Carr, have joined the DJs in the studio to show their support.
Moyles previously told DS that the record attempt could be "the witnessing of an absolute physical and mental breakdown".
> Click here to read our recent interview with Chris Moyles about the record attempt