The painting is the centre piece of an exhibition on the Spanish artists active in the southern Italian city during the Renaissance, which opened at the Capodimonte Museum on Monday.
Commissioned in around 1514 for the Doce family chapel in the church of San Domenico Maggiore in Naples, the altarpiece featuring Mary, the baby Jesus, Archangel Raphael, Tobias and Saint Jerome, was subsequently removed by Spanish rulers and transferred to Madrid around the mid-17th century.
It is now housed in the Prado Museum in the Spanish capital, which has helped put the exhibition together.
The Spanish in Naples. The Southern Renaissance presents a selection of works executed by some of the leading Spanish artists active in Naples during the first 30 years of the 16th century.
The artists include Pedro Fernández, Bartolomé Ordóñez, Diego de Siloe, Pedro Machuca and Alonso Berruguete, with special attention to the close connection between the sister disciplines of painting and sculpture.
"Naples not only has great museums, but it is (itself) a great scattered museum, making the city unique in the world," Culture Minister Gennaro Sangiuliano said at the opening of the exhibition.
"Naples is experiencing an extraordinary cultural season, and Spain, which is at home in Naples, wants to make a strong contribution," the Spanish ambassador to Italy, Miguel Ángel Fernández-Palacios Martínez, said.
The exhibition runs until June 25.
ANSA