Markers linked to QTLs are useful in practical plant breeding, only if they get validated in genotypes of independent populations and diverse genetic backgrounds. 41 SSR markers reported linked to QTLs for grain iron (Fe) and zinc (Zn) concentration in wheat were analysed. Only 16 of them showed polymorphism and the remaining 25 turned out to be monomorphic in 48 wheat genotypes used in the present study. Single marker analysis (SMA) for the 16 polymorphic markers was carried out to assess the linkage between marker and the trait, based on which two markers (Xbarc186 and Xbarc74) for grain Fe concentration and three markers (Xgwm3, Xwms149 and Xgwm538) for grain Zn concentration were validated in the present study. The phenotypic variations explained by Xbarc186, Xbarc74, Xgwm3, Xwms149 and Xgwm538 were 40.2% and 19.8%, 10.7%, 21.7%, and 39.6%, respectively. The validation of these SSRs may be useful in breeding wheat with high grain Fe and Zn concentration.