The centrepiece of the National Gallery of Victoria's winter masterpieces exhibition will be Raphael's The Holy Family with St John.

The work, also known as Madonna of the Rose, comes from Spain's Museo del Prado collection of works that will be exhibited at NGV International as part of this year's survey of Italian masters.

Premier Denis Napthine said the work had left the Prado only once before. It is one of 70 paintings and 30 drawings from masters such as Titian and Tiepolo.

''It's an important work because it was done by one of the most influential painters of Western art,'' said Miguel Falomir Faus, curator of the exhibition from the Museo del Prado. ''But more importantly, this was executed by Raphael himself. In the last year of his life, many of Raphael's paintings were executed by his assistants, so this is a very important painting,'' he said.

''The exhibition spans three centuries and gives a comprehensive approach to the history of Italian paintings,'' he said.