Honda rider Joan Mir topped the session as he recorded the fastest time in the final Barcelona MotoGP session.
The Spaniard posted a 1.39.425s, lapping over two tenths clear of Pedro Acosta, Fabio Di Giannantonio and Jorge Martin.
Fabio Quartararo and Yamaha finished in fifth spot, meaning all top five positions were covered by all five manufacturers.
Franco Morbidelli and Alex Marquez followed, claiming sixth and seventh in the Ducati satellite teams.
Raul Fernandez and Francesco Bagnaia lapped eighth and ninth fastest, followed by Enea Bastianini.
The championship leader, Marco Bezzecchi, followed in 11th, with Diogo Moreira and Brad Binder close behind.
Fermin Aldeguer and Johann Zarco posted 14th and 15th, with Maverick Vinales on his return in 16th place.
Despite Mir topping the session, his team-mate Luca Marini finished 17th on the timesheet, ahead of Pramac duo Toprak Razgatlioglu and Jack Miller.
Ai Ogura followed, before Yamaha test rider Augusto Fernandez claimed 21st, with Alex Rins rounding off proceedings.
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