In the late 1890s he worked in Cambridge, Massachusetts and showed his paintings in New York at the Society of American Artists and also in Boston.
He returned to Europe in 1901, where he visited Spain to study the painting of Velázquez and El Greco and traveled through Brittany, and the Netherlands to see portraits by his "heroes", Frans Hals and Rembrandt.
Adopting the colour theories of his former neighbour Denman Ross, who had become a prominent collector and a teacher at Harvard, Hopkinson later used the results of Carl Cutler's experiments with a spinning disk to study the color spectrum.
Instead of allying himself with the local established painters, Hopkinson showed his work with the "Boston Five", a group of young watercolorists though he continued to paint in oil for an elite clientele.
In the mid-1920s, Hopkinson took on a young Boston painter Pietro Pezzati as his assistant, who worked with him at his Fenway studio.