He owned the Italian supercar for just two years before selling it to its current owner, an Austria-based motoring enthusiast, in 2009.
Several pictures remain of the Maggie May singer with the blue convertible motor, including a number which were taken outside his lavish home.
In the shots, Stewart, wearing a Celtic FC hoodie, it stood in between the Gallardo and a red Ferrari, which he also owned.
It is thought he also owned an identical Gallardo in America.
As a known collector of high-performance vehicles, it is of little surprise that Sir Rod didn’t use the car on a daily basis, racking up just 2,243 miles during his ownership.
The car has continued to be used sparingly in the years since and now has just over 3,000 miles on the clock.
The car, that is powered by a 5 litre V10 engine and has a top speed of 196mph, is now being sold through auctioneers RM Sotheby’s.
Augustin Sabatie-Garat, auction manager at RM Sotheby’s, said: “Stewart has always been well known for his supercar collection, and this Lamborghini is a stand-out example in the musician’s list of exotic cars owned.
“The Gallardo Spyder was delivered new to Stewart, with invoices included in the sale to confirm this, as the pop legend chose the highly unique colour scheme over a Blu Scylla and interior.
“Aside from the celebrity connection, the car is a highly desirable example of a Gallardo Spyder in its own right.”
The auctioneers also added the car was a fantastic opportunity for any hardcore Rod Stewart fans.
A spokesman said: “It was only natural that Stewart, a Lamborghini collector and enthusiast for decades, wanted to own the latest and greatest that they had to offer.
“Beautifully maintained and lovingly preserved from new, this Gallardo Spyder is an exceptionally rare and exciting proposition, not only thanks to its celebrity ownership, but also to the fact that it still wears its original paint and has never been involved in an accident.
“With the vast majority of mileage accrued by Stewart himself, this would be a thrilling purchase for any fan of Rod Stewart or Lamborghini enthusiast.
“It would be difficult to find a Gallardo Spyder worldwide presented in such exceptional condition with the added bonus of spending time in Rod Stewart’s garage.”
Also in the same sale is another of Stewart’s Lamborghinis – a 1971 Miura.
The sale takes place on September 5 in London.