The effects of 16 herbicides from 14 different chemical classes on levels of soluble protein, hydroxyphenolics, anthocyanin, and chlorophyll were determined in light- and dark-grown soybean [Glycine max (L.) Merr. ‘Hill’] seedlings. Growth-reducing concentrations of the herbicides were supplied to 3-day-old dark-grown soybean seedlings in liquid culture. Soluble protein (per axis) was reduced by fluridone {1-methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone}, paraquat (1,1′-dimethyl-4,4′-bipyridinium ion), perfluidone {1,1,1-trifluoro-N-[2-methyl-4-(phenylsulfonyl)phenyl] methanesulfonamide}, and propanil (3′,4′-dichloropropionanilide) 24 or 48 h after treatment. In light-grown plants, soluble hydroxyphenolic compound levels were decreased on a per axis basis after 48 h by all chemical treatments except by atrazine [2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine], the methyl ester of diclofop {2-[4-(2,4-dichlorophenoxy)phenoxy] propanoate}, DSMA (disodium methanearsonate), fluridone, MH (1,2-dihydro-3,6-pyridazinedione), nitralin [4-(methylsulfonyl)-2,6-dinitro-N,N-dipropylaniline], TCA (trichloroacetic acid), and 2,4-D [(2,4-dichlorophenoxy)acetic acid]. Total chlorophyll content in hypocotyls of these seedlings was decreased by fluridone, metribuzin [4-amino-6-tert-butyl-3-(methylthio)-as-triazin-5 (4H)-one], norflurazon [4-chloro-5-(methylamino)-2-(α,α,α-trifluoro-m-tolyl)-3 (2H)-pyridazinone], paraquat, and 2,4-D, but others had no significant effect. Anthocyanin accumulation in hypocotyls of 48-h light-grown seedlings was decreased by atrazine, fenuron (1,1-dimethyl-3-phenylurea), metribuzin, norflurazon, paraquat, propanil, and propham (isopropyl carbanilate). Phenylalanine ammonia-lyase (PAL) activity (previously reported work) was positively correlated with anthocyanin levels in tissues after treatment with these 16 herbicides, but not with glyphosate [N-(phosphonomethyl)glycine]. There was, however, no correlation between extractable PAL activity and chlorophyll, soluble hydroxyphenolic compounds, or soluble protein content. These results indicated that extracted PAL activities usually reflect relative in vivo activities and that PAL activity is limiting to phenylpropanoid synthesis in vivo.