Multiple stable isotope ratios (δ2H, δ13C, δ18O and δ34S) were measured in muscle, muscle lipids and lipid fractions collected from 28 lambs, subjected to a diet-switch and raised on two energy allowances (EAs), to determine tissue turnover and diet-tissue fractionation. The diet-muscle fractionations prior to the diet-switch were estimated to be -44.0‰, +1.9‰ and 0‰ for H, C and S, respectively, while the drinking water was demonstrated to be the main source of muscle O and thus δ18O variation. The diet-intra-muscular lipid fractionations prior to the diet-switch were estimated to be -172.7‰, -1.3‰ and -11.5‰ for H, C and O, respectively. The C half-lives of muscle were determined to be 75.7 and 91.6days for animals receiving the high and low EA, respectively. Extracting temporally resolved pre-slaughter dietary information from meat by analysing bulk muscle, muscle lipids and muscle lipid fractions appeared to be not practicable due to possible incomplete turnover of lipids.