They were beaten 7-6, 6-4 by Venus’ former partner, South African Raven Klaasen, and New Zealand-born Ben McLachlan, who hails from Queenstown but represents Japan.
It was certainly a match that got away from the second seeds.
Venus and Skupski were up a break in the first set but could not close it out.
They converted just one of nine break-point chances with Klaasen and McLachlan converting twice from five chances.
McLachlan, who has now won seven ATP doubles titles, said he was thrilled with the win.
Venus will now travel to Toronto for the ATP Masters 1000 tournament, reuniting with new regular partner Tim Puetz, of Germany.
They are the seventh seeds and have been drawn against the Belgian pair of Sander Gille and Joran Vliegen in the first round.
