<i>Aim:</i> To assess longitudinally whether smoking tobacco is related to muscle strength in healthy adults and to assess the influence of lifestyle covariates on this relation. <i>Methods:</i> Data were obtained from the observational Amsterdam Growth and Health Longitudinal Study, with four repeated measurements at the ages of 21 (n = 181), 27 (n = 144), 32 (n = 426) and 36 (n = 373) years. Knee muscle strength (KMS) was measured at an angle of 300° per second. Smoking and the covariates physical activity and dietary intake were assessed with a questionnaire. Other covariates included physical fitness and body fat percentage. Cardiopulmonary fitness was assessed with a maximal running test and body fatness with skinfold measurements. Sex-specific longitudinal analyses were carried out using generalized estimating equations. <i>Results:</i> In men and women, an inverse relationship between smoking tobacco and KMS was found. Smoking 100 g of tobacco a week resulted in a reduction of 2.9% in KMS in men and a reduction of 5.0% in women. This relation existed largely independently of lifestyle covariates.<i> Conclusion:</i> This study indicates that smoking is inversely related to skeletal muscle strength in healthy adults.