One and two dimensional NMR spectroscopy has been employed to characterize the various metabolites of serum control healthy samples. Two dimensional heteronuclear experiment has been included to take advantage of larger chemical shift spread of ¹³C resonances allowing a more detailed identification of metabolites not possible in one dimensional spectra. This methodology has successfully allowed the assignment of ninety four resonances of various metabolites. The importance of the work lies in the fact that characteristic fingerprints of various metabolites of serum samples of normal healthy control have been obtained which can identify and distinguish metabolic differences from other diseased specimens or non-diseased/healthy serum samples. This study will help enhance the literature of metabolite identification in serum samples.