Amphiphilic lipids and hydrophobic proteins are vaporized at atmospheric pressure using nonresonant 70 femtosecond (fs) laser pulses followed by electrospray post-ionization prior to being transferred into a time-of-flight mass spectrometer for mass analysis. Measurements of molecules on metal and transparent dielectric surfaces indicate that vaporization occurs through a nonthermal mechanism. The molecules analyzed include the lipids 1-monooleoyl-rac-glycerol, 1,2-dihexanoyl-sn-glycero-3-phosphocholine, 1,2-dimyristoyl-sn-glycero-3-phosphocholine, and the hydrophobic proteins gramicidin A, B, and C. Vaporization of lipids from blood and milk are also presented to demonstrate that lipids in complex systems can be transferred intact into the gas phase for mass analysis.