Single-conformation spectroscopy of the three lignin monomers (hereafter "monolignols") p-coumaryl alcohol (pCoumA), coniferyl alcohol (ConA), and sinapyl alcohol (SinA) has been carried out on the isolated molecules cooled in a supersonic expansion. Laser-induced fluorescence excitation, dispersed fluorescence, resonant two-photon ionization, UV-UV hole-burning, and resonant ion-dip infrared spectroscopy were carried out as needed to obtain firm assignments for the observed conformers of the three molecules. In each case, two conformers were observed, differing in the relative orientations of the vinyl and OH substituents para to one another on the phenyl ring. In pCoumA, the two conformers have S(0)-S(1) origins nearly identical in size, split from one another by only 7 cm(-1), in close analogy with previous results of Morgan et al. on p-vinylphenol ( Chem. Phys. 2008 , 347 , 340 ). ConA, with its methoxy group ortho to the OH group, also has two low-energy conformers forming a syn/anti pair, in this case with the OH group locked into an orientation in which it forms an intramolecular H-bond with the adjacent methoxy group. The electronic frequency shift between the two conformers is dramatically increased to 805 cm(-1), with the dominant conformer of ConA (with S(0)-S(1) origin at 32 640 cm(-1)) about 5 times the intensity of its minor counterpart (with S(0)-S(1) origin at 33 444 cm(-1)). The presence of an OH···OCH(3) intramolecular H-bond is established by the shift of the OH stretch fundamental of the OH group to 3599 cm(-1), as it is in o-methoxyphenol ( Fujimaki et al. J. Chem. Phys. 1999 , 110 , 4238 ). Analogous single-conformation UV and IR spectra of o-methoxy-p-vinylphenol show a close similarity to ConA and provide a basis for a firm assignment of the red-shifted (blue-shifted) conformer of both molecules to the syn (anti) conformer. The two observed conformers of SinA, with its two methoxy group straddling the OH group, have S(0)-S(1) origins split by 239 cm(-1) (33 055 and 33 294 cm(-1)), a value between those in pCoumA and ConA. A combination of experimental data and calculations on the three monolignols and simpler derivatives is used to establish that the conformational preferences of the monolignols reflect the preferences of each of the ring substituents separately, enhanced by the presence of the intramolecular OH···OCH(3) H-bond. Taken as a whole, the presence of multiple flexible substituents locks in certain preferred orientations of the groups relative to one another, even in the apparently flexible allyl alcohol side chain (-CH═CH-CH(2)OH), where the OH group orients itself so that the hydrogen is pointed back over the vinyl π cloud in order to minimize interactions between the oxygen lone pairs and the π electrons.