The painting collection possesses more than three hundred and forty works by old masters created from the end of the 15th to the second half of the 19th century. Among them, a particular place is occupied by a smallish collection of paintings from the workshops of leading Dalmatian and Dubrovnik artists of the 15th and 16th centuries. One such work is the Polyptych of Our Lady made in the works of Blaž son of Juraj of Trogir in the spirit of the late Venetian trecento.

Exhibited in the permanent display of the museum are works of Lovro Dobričević, St Blaise, and Mihajlo Hamzić, The Baptism of Christ. They were both trained in Italy, and on their return to Dubrovnik opened up their own workshops, creating a particular style that links the new Renaissance trends with the principles of late Gothic painting.

From the same period are the works of foreign, mainly Italian, artists like Vicenzo Catena, Girl and Flute Player, Paris Bordone, Venus and Adonis, with the occasional work of central European masters.
Most of the 17th and 18th century paintings are also from Italy, with a few exceptions that derive from Germany and Dubrovnik.

The collection has more than 100 portraits of Dubrovnik figures, leading scholars and scientists, writers and politicians ( such as Bošković, Gradi, Ghetaldi, Gondola, Bona, Caboga and Bassegli ), the authors of which are lesser known or anonymous painters.