Barnabe Barnes attempted, in 1598, to kill the recorder of Berwick with poisoned claret, but he successfully evaded sentence. He published a sonnet sequence, Parthenophil and Parthenophe. Sonnets, madrigals, elegies, and odes (1593), notable as one of the first of such collections to appear after Sidney's Astropher and Stella. Sonnet #54 clearly states his bisexuality.

He also wrote A divine centurie of spirituall sonnets (1595), Foure bookes of offices (1606), and The divils charter: a tragaedie conteining the Lie and Death of Pope Alexander the Sixt (1607), a vigorous Machiavellian drama which includes such melodramatic scenes as the murder of Lucrezia Borgia with poisoned face wash. Barnes' poetry is remarkable for its vigour and technical range.