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Paleoenvironmental and paleoclimatic variations experienced by the Mediterranean region during the Miocene Burdigalian-Langhian interval have been investigated through analysis of biomarker signals (n-alkanes and glycerol dialkyl glycerol tetraethers, GDGTs) from slope mudstones of the northern Apennines (Marne di Vicchio Fm). Terrestrial signatures in marine sediments reflected by preserved long-chain n-alkanes with a identifiable odd-over-even carbon number predominance attest to the thermal immaturity of the sedimentary rock. Abundances of n-C27, n-C29 and n-C31 alkanes, coupled with the carbon isotopic signatures of the n-C31 homologue suggest that Mediterranean vegetation crossed a transition phase adapting to arid conditions that may favored a shift from woodland to wooded grassland during the Middle Miocene. This ecosystem evolution could have been favored by the passage from warm humid conditions to a warm drier climate, as indicated by GDGT-based TEX86 and BIT indexes. Strong similarities between these Mediterranean biomarker records and those from the Atlantic (DSDP Sites 94, 516, 608) imply a constant connection of the western Mediterranean to the Atlantic Ocean from ~19 Ma to ~14.5 Ma.